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Second Edition
Handbook of
Plant-Based
Fermented Food and
Beverage Technology
Second Edition
Handbook of
Plant-Based
Fermented Food and
Beverage Technology
Edited by Y. H. Hui
Administrative Associate Editor
E. Özgül Evranuz
Associate Editors
Francisco Noé Arroyo-López
Lihua Fan
Åse Solvej Hansen
M. Eugenia Jaramillo-Flores
Marica Rakin
Rosane Freitas Schwan
Weibiao Zhou
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
The previous edition was published with a different title: Handbook of Food and Beverage Fermentation Technology (ISBN
9780824747800)
CRC Press
Taylor & Francis Group
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Boca Raton, FL 33487-2742
© 2012 by Taylor & Francis Group, LLC
CRC Press is an imprint of Taylor & Francis Group, an Informa business
No claim to original U.S. Government works
Version Date: 20120425
International Standard Book Number-13: 978-1-4398-7069-3 (eBook - PDF)
This book contains information obtained from authentic and highly regarded sources. Reasonable efforts have been made to
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Contents
Preface....................................................................................................................................................... ix
Editors........................................................................................................................................................ xi
Contributors.............................................................................................................................................xiii
Part I
Introduction
1. Fermented Plant Products and Their Manufacturing.................................................................. 3
Y. H. Hui
2. Flavors and Food Fermentation..................................................................................................... 23
Shao Quan Liu
3. Fermentation and Biopreservation of Plant-Based Foods with Lactic Acid Bacteria.............. 35
Lihua Fan and Lisbeth Truelstrup Hansen
4. Plant-Based Fermented Foods and Beverages of Asia................................................................. 49
Jyoti Prakash Tamang
Part II
Soy Products
5. Soy Sauce: Typical Aspects of Japanese Shoyu and Indonesian Kecap.................................... 93
Hanifah Nuryani Lioe, Anton Apriyantono, and Masaaki Yasuda
6. Synbiotic Soy Beverages: Principles and Sensory Attributes................................................... 103
Olga Lucía Mondragón-Bernal, Francisco Maugeri Filho,
José Guilherme Lembi Ferreira Alves, and Maria Isabel Rodrigues
7. Thua Nao: A Traditional Thai Fermented Soy Product............................................................131
Ekachai Chukeatirote, Katekan Dajanta, and Arunee Apichartsrangkoon
8. Soymilk and Tofu Manufacturing............................................................................................... 139
Sam K. C. Chang and Zhisheng Liu
Part III
Fruits and Fruit Products
9. Sensory Analysis of Fruit and Fermented Fruit Product Flavors............................................165
Soo-Yeun Lee, Lisa Allgeyer, Erika Neely, and Joseph Kreger
10. Wine Fermentation and Production.............................................................................................179
Juan José Rodríguez-Bencomo, M. Ángeles Pozo-Bayón, and M. Victoria Moreno-Arribas
11. Fermentation of Caper Products................................................................................................. 201
Rubén Pérez Pulido, Nabil Benomar, Magdalena Martínez Cañamero, Hikmate Abriouel,
and Antonio Gálvez
v
vi
Contents
12. Apple Cider Fermentation............................................................................................................ 209
Alessandro Nogueira and Gilvan Wosiacki
13. Fermentation and Cashew Apple Juice....................................................................................... 237
Ana Lúcia Fernandes Pereira and Sueli Rodrigues
14. Production and Characterization of Wine from Mango (Mangifera indica L.) Fruit Juice......... 249
Lebaka Veeranjaneya Reddy and Obulam Vijaya Sarathi Reddy
15. Stone Fruit: Wine and Brandy.................................................................................................... 273
Vinod Kumar Joshi, Rakesh Sharma, and Ghan Shyam Abrol
16. Fermentation of Olive Fruit......................................................................................................... 307
Francisco Noé Arroyo-López, Joaquín Bautista-Gallego, Verónica Romero-Gil,
Jose María Baquero, Pedro García-García, Rufino Jiménez-Díaz, Antonio López-López,
Francisco Rodríguez-Gómez, and Antonio Garrido-Fernández
17. Noni Fruits..................................................................................................................................... 327
Scot Nelson
18. Wine Production in the United States: Regulatory Requirements.......................................... 339
Y. H. Hui
Part IV
Vegetables and Vegetable Products
19. Vegetable Fermentation with Economical Significance in South America: Use of
Ensilage as a Probiotic Vehicle..................................................................................................... 363
Mario Eduardo Arena, Ana Lidia Apás, and Silvia Nelina González
20. Fermented Red Beet Juice............................................................................................................ 373
Zsolt Zalán, Anna Halász, and Ágnes Baráth
21. Health Benefits of Fermented Vegetable Juices.......................................................................... 385
Marica Rakin, Maja Vukašinović Sekulić, and Ljiljana Mojović
22. Almagro Eggplant: From Homemade Tradition to Small-Scale Industry............................. 407
Susana Seseña and María Llanos Palop
23. Olives in Commerce in the United States....................................................................................419
Y. H. Hui
24. Pickle Manufacturing in the United States: Quality Assurance and Establishment
Inspection........................................................................................................................................435
Y. H. Hui
25. Canned Sauerkraut: Manufacturing, Inspection, and Grade Standards................................451
Y. H. Hui
26. Mexican Jalapeño Pepper: Properties, Manufacture, and Flavor........................................... 463
Karendash González-Quijano, Lidia Parada-Dorantes, Lidia Dorantes-Alvarez,
Humberto Hernández-Sánchez, and M. Eugenia Jaramillo-Flores
vii
Contents
Part V
Cereals and Cereal Products
27. Fermented Bread........................................................................................................................... 477
Weibiao Zhou and Nantawan Therdthai
28. Sourdough Bread.......................................................................................................................... 493
Åse Solvej Hansen
29. Liquid Sourdough Fermentation..................................................................................................517
Paola Carnevali, Fabio Minervini, and Aldo Corsetti
30. Chinese Fermented Rice Noodles................................................................................................ 527
Yong Yang
31. Boza: A Traditional Cereal-Based, Fermented Turkish Beverage...........................................533
Sirma Yegin and Marcelo Fernández-Lahore
32. Chinese Steamed Buns.................................................................................................................. 543
Suwimon Keeratipibul and Naphatrapi Luangsakul
33. Whiskey Manufacture...................................................................................................................557
Y. H. Hui
34. Beer Production in the United States: Regulatory Requirements........................................... 565
Y. H. Hui
35. Distilled Spirits Production in the United States: Regulatory Requirements........................ 579
Y. H. Hui
Part VI
Specialty Products
36. Traditional Balsamic Vinegar: A Microbiological Overview................................................... 609
Lisa Solieri, Maria Gullo, and Paolo Giudici
37. Palm Wine.......................................................................................................................................631
Maurizio Ciani, Marzia Stringini, and Francesca Comitini
38. Brazilian Cachaça: Fermentation and Production.................................................................... 639
Fernanda Badotti, Fátima C. O. Gomes, and Carlos A. Rosa
39. Brick Tea........................................................................................................................................ 649
Haizhen Mo and Xinhong Liang
40. Shalgam (Şalgam).......................................................................................................................... 657
Huseyin Erten and Hasan Tanguler
41. Fermented Coconut Milk and Coconut Oil................................................................................ 665
Azila Abdul Aziz, Ramlan Aziz, Muhammad Roji Sarmidi, Chua Lee Suan,
Nur Arbainah Shamsul Annuar, Norhayati Mohamed Noor, and Norfarahiyah Aman Nor
viii
Contents
42. Coffee Fermentation..................................................................................................................... 677
Rosane Freitas Schwan, Cristina Ferreira Silva, and Luis Roberto Batista
43. Pulque Fermentation......................................................................................................................691
Adelfo Escalante, Martha Giles-Gómez, Guadalupe Esquivel Flores, Violeta Matus Acuña,
Rubén Moreno-Terrazas, Agustín López-Munguía, and Patricia Lappe-Oliveras
44. Probiotic Nondairy Beverages..................................................................................................... 707
Carlos Ricardo Soccol, Juliano De Dea Lindner, Caroline Tiemi Yamaguishi,
Michele Rigon Spier, Luciana Porto de Souza Vandenberghe, and Vanete Thomaz Soccol
Part VII
Fermentation and Food Ingredients
45. Fresh and Fermented Vegetables as a Source of Proteolytic Bacteria......................................731
Catalina Elena Kotlar, María Victoria Agüero, Alejandra Graciela Ponce,
Olinda Pérez Borla, and Sara I. Roura
46. Soymilk Fermentation and Enzymes Production.......................................................................757
Marisa S. Garro and Graciela Savoy de Giori
47. Microorganisms and Microbially Derived Ingredients Used in Foods.................................... 771
Y. H. Hui
Preface
Fermented food is a very interesting category of food products. Not only can it be produced with inexpensive ingredients and simple techniques, but it also makes a significant contribution to human diet,
especially in rural households and village communities in many areas of the world. In every ethnic group
in the world, there are fermented foods produced from recipes handed down from generation to generation. Such food products play an important role in cultural identity, local economy, and gastronomical
delight. With progresses in biological and microbiological sciences, the manufacture of some of the more
popular fermented food products has been commercialized, while others are still produced at home or in
small-scale industries using traditional methods.
Fermentation changes the initial characteristics of a food into a product that is significantly different
but highly acceptable by consumers. Of course, consumer preference for fermented food varies within
and between cultures. For example, within the United States, many consumers like sauerkraut, although
some do not. The trend in North America is toward acceptance and preference of foreign fermented nonEuropean food products. You can find Chinese fermented black beans and Korean Kim Chee in most
Asian markets and some major grocery chains in North America.
Although reference books on fermented foods have been in existence for at least 50 years, those with
details on the science, technology, and engineering of food fermentation began to appear after 1980.
Scientific literature in the past decade has been flooded with new applications of genetic engineering in
the fermentation of food products, especially in the breeding of plants.
This book provides an up-to-date reference for fermented foods and beverages derived from plant
products. Almost every book on food fermentation has something not found in others. The Handbook
of Plant-Based Fermented Food and Beverage Technology provides a detailed background of history,
microorganisms, quality assurance, and the manufacture of general fermented food products and discusses the production of six categories of fermented foods and beverages:
•
•
•
•
•
•
Soy: soy sauce, tofu
Fruit: wine, olive
Vegetables: red beet juice, pickles
Cereals: sourdough, beer
Specialties: tea, coffee
Ingredients: enzymes, probiotic bacteria
Traditional fermented products are discussed, including wine, sourdough, and olive. We also present
details of the manufacture and quality characteristics of some fermented foods that are less common in
other books on fermentation in the English language: fermented products from East Asia, for example,
soy sauce, and fermented cereal products from Turkey, for example, boza. Although this book has covered a variety of products, many topics are omitted for different reasons including space limitation,
product selection, and the contributors’ areas of expertise.
This book is unique in several aspects: it is an updated and comprehensive reference source, it contains
topics not covered in similar books, and its contributors include experts from government, industry, and
academia worldwide. The book has 47 chapters and is divided into seven parts. It is the cooperative effort
of more than 60 contributors from more than 20 countries (Argentina, Brazil, Canada, China, Denmark,
Germany, Himalaya, Hungary, India, Indonesia, Italy, Japan, Malaysia, Mexico, Serbia, Singapore,
Spain, Thailand, Turkey, and the United States) with expertise in one or more fermented products, led by
an editorial team of nine academicians from seven countries (Brazil, Canada, Denmark, Mexico, Serbia,
ix
x
Preface
Singapore, Spain, Turkey, and the United States). The seven associate editors of this team are all experts
in their areas of specialization as indicated below:
• Dr. Francisco Noé Arroyo-López is a research scientist with the Department of Food
Biotechnology, Instituto de la Grasa (CSIC), Seville, Spain. His expertise focuses on table olives
and fermented vegetables, as supported by his published work in journals and book chapters.
• Dr. Lihua Fan is a research scientist with Agriculture and Agri-Food Canada. Food fermentation and microbial safety of fruits and vegetables are her major research interests, among
others, as documented by her numerous published journal articles, book chapters, and other
professional works.
• Professor Åse Solvej Hansen is with the Department of Food Science, University of Copenhagen.
Her major research has focused on rye and wheat baking technology and microbiology, including the influence of sourdough fermentation on bread quality, as documented by her extensive
publications.
• Dr. M. Eugenia Jaramillo-Flores is with the Department of Food Science, National Polytechnic
Institute, Mexico. With an expertise in food biochemistry and biotechnology, she focuses her
research on nutraceuticals and functional foods, as confirmed by her published work in peerreviewed journals and other professional publications that are nationally and internationally
recognized.
• Dr. Marica Rakin is with the Department of Biochemical Engineering and Biotechnology,
University of Belgrade, Serbia and Montenegro. She has published extensively on fermentation, and her research focuses on the production of bio-fermented vegetables and drinks and the
production of bio-ethanol related to renewable biomass.
• Dr. Rosane Freitas Schwan is an agronomist at Federal University of Lavras, Minas Gerais,
Brazil, where she is mostly involved in research on microbiology of several fermented foods
including cocoa and coffee. In addition, she is studying the development of culture starters to
improve the quality of fermentative processes. • Dr. Weibiao Zhou is the director of the Food Science and Technology Program at the National
University of Singapore. Dr. Zhou’s research interests and expertise are in food engineering
and food processing, particularly bakery products, as supported by his numerous publications.
In sum, the approach for this book makes it an essential reference on plant-based fermented foods and
beverages. The editorial team thanks all the contributors for sharing their experience in their fields of
expertise. They are the people who made this book possible. We hope you enjoy and benefit from the
fruits of their labor.
We know how hard it is to develop the content of a book. However, we believe that the production of a
professional book of this nature is even more difficult. We thank the production team at CRC Press. You
are the best judge of the quality of this book.
Y. H. Hui
United States
E. Özgül Evranuz
Turkey
Editors
Y. H. Hui received his PhD from the University of California at Berkeley. He is currently a senior scientist with the consulting firm of Science Technology System at West Sacramento, California. He has
authored, coauthored, edited, and coedited more than 30 books in food science, food technology, food
engineering, and food laws, including Handbook of Food Product Manufacturing (2 volumes, Wiley,
2007) and Handbook of Food Science, Technology, and Engineering (4 volumes, CRC Press, 2006). He
has guest-lectured in universities in the Americas (North, Central, and South), Europe, and Asia and is
currently a consultant for the food industries, with an emphasis on food sanitation and food safety.
E. Özgül Evranuz is a professor (1994–present) of food processing at the Food Engineering Department
of İstanbul Technical University (ITU), Turkey. After graduating from the Chemical Engineering
Department of Middle East Technical University (METU; 1971), Ankara, Turkey, she has conveyed her
academic career in the area of food technology. She holds MS degrees in Chemical Engineering (METU)
and Food Science and Management (University of London, Queen Elizabeth Collage, London) and a
PhD in Chemical Engineering (ITU). She is among the builders of the Food Engineering Department of
ITU. Her main interests are preservation, storage, and packaging of food plant materials. She has served
as an editor of a book and has several publications in the field.
Associate Editors
Francisco Noé Arroyo-López received a degree in biology from the University of Jaen, Spain (2001),
where he also received his PhD. His doctoral thesis at the University of Seville (Spain) was about
table olive fermentations (2007). Currently, he is a postdoctoral researcher at the Department of Food
Biotechnology from the Instituto de la Grasa (CSIC). His areas of research are mainly application of
mathematical models to predict microorganism growth/survival in table olive processing and the use of
molecular methods for identification of lactic acid bacteria, yeasts, and Enterobacteriaceae related to
this fermented vegetable. He has more than 40 international publications in peer-reviewed SCI journals
as well as 6 chapter books. He is a professor in diverse international table olive–related courses.
Lihua Fan is a research scientist working at the Atlantic Food and Horticulture Research Centre,
Agriculture and Agri-Food Canada, since 1998. She received her MS and PhD in food science from
China Agricultural University in 1989 and 1991, respectively. Her research interests include natural
antimicrobials on safety and quality of whole and fresh-cut fruit and vegetables, bioactive compounds
and biopreservatives, food fermentation, and disease control of postharvest horticultural produce. Fan is
a member of the International Association for Food Protection and a member and subpanel member of
the Food Microbiology Division of the Institute of Food Technologists. She is a committee member of
the Multi-State Research Project S-294 for the National Information Management Support System, USA,
and a Graduate Supervisory Committee member in Food Program of Dalhousie University, Canada. In
recognition of her supervising efforts, she recently received the Canadian Forces Liaison Council’s Nova
Scotia Provincial Award for Best Practices in Employer Support.
xi
xii
Editors
Åse Solvej Hansen is an associate professor of the Department of Food Science, University of
Copenhagen, Denmark. She received her MSc in food science and technology from the Royal Veterinary
and Agricultural University, Denmark, and has worked with quality control and product development for
several years in a Danish industrial bakery. With a background in practical problems from the baking
industry, she has worked with cereal research focusing on wheat and rye baking technology, including
the influence of sourdough fermentation on bread quality. Her bread quality research has also included
the formation of flavor compounds, phytate degradation, and microbial stability of sourdoughs.
M. Eugenia Jaramillo-Flores received her BS in biochemical engineering and MS and PhD in food
at the National School of Biological Sciences, National Polytechnic Institute, Mexico. She is currently
a professor of food biochemistry and the chair of the PhD program in biotechnology. She has directed
undergraduate and graduate theses on food science and food biotechnology. She belongs to different
international scientific associations of chemical, food, and pigments (Institute of Food Technologists,
American Chemical Society, and International Carotenoids Society). She has participated in countless
international conferences and has published articles in both national and international journals. She
works as a researcher in the area of nutraceuticals and functional foods, particularly in the evaluation of
the effect of the process.
Marica Rakin is an associate professor since December 2011 at the Department of Biochemical
Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade,
Belgrade, Serbia, where she received PhD. Her thesis was titled “The Obtaining of Bioactive Materials
on Basis Brewer’s Yeast and Healing Vegetables.” She has extensive experience in the application and
development of novel by-products of food in agroindustry, with an emphasis on the technological and
microbiological aspects of the production of biofermented vegetables and drinks and the bioethanol production on renewable biomass. She has special expertise in yeast (Saccharomyces cerevisiae) fermentation and lactic acid bacteria with an emphasis on their applications.
Rosane Freitas Schwan is an agronomist and associate professor in microbiology and has been working
for more than 15 years at the Federal University of Lavras, Minas Gerais, Brazil, where she is mostly
involved in research on microbiology of several fermented foods. She worked for 12 years at Cocoa
Research Centre in Itabuna, Bahia, Brazil, where she developed a microbial cocktail for cocoa fermentation. Her research interest includes biodiversity and phylogeny of bacteria, yeasts, and filamentous fungi
involved in cocoa, coffee, and other tropical fermentations and biodiversity of yeasts in agrofood ecosystems and development of culture starters to improve the quality of fermentative processes.
Weibiao Zhou is a full professor and the director of the Food Science and Technology Program at the
National University of Singapore. He is a board member of the Agri-food and Veterinary Authority of
Singapore, a fellow of the Australian Institute of Food Science and Technology, a council member of
the Singapore Institute of Food Science and Technology, a member of the Singapore Food Standards
Committee, and a member of the editorial boards of five international peer-refereed journals in the area
of food engineering and food processing. Educated in China and Australia, he has held various professional positions in a number of organizations in Australia, Canada, China, France, and the United States.
With more than 180 scientific publications, Zhou’s research interests and expertise are in food engineering and food processing, particularly baking processes, dairy processes, drying, functional foods, and
process modeling, optimization, and control.
Contributors
Hikmate Abriouel
Departamento de Ciencias de la Salud
Universidad de Jaén
Jaén, Spain
Arunee Apichartsrangkoon
Department of Food Science and Technology
Chiang Mai University
Chiang Mai, Thailand
Ghan Shyam Abrol
Department of Food Science and Technology
Dr. Y. S. Parmar University of Horticulture and
Forestry
Solan, India
Anton Apriyantono
Pagelaran Ciomas
Bogor, Indonesia
Violeta Matus Acuña
Departamento de Ingeniería Celular y Biocatálisis
Instituto de Biotechnología
Universidad Nacional Autónoma de México
Cuernavaca, Morelos, México
María Victoria Agüero
Departamento de Ingeniería Química y en
Alimentos
Universidad Nacional de Mar del Plata
Mar del Plata, Argentina
and
Consejo Nacional de Investigaciones Científicas
y Técnicas
Buenos Aires, Argentina
Lisa Allgeyer
Department of Food Science and Human Nutrition
University of Illinois
Urbana-Champaign, Illinois
José Guilherme Lembi Ferreira Alves
Food Science Department
Federal University of Lavras
Lavras, Minas Gerais, Brazil
Ana Lidia Apás
Facultad de Bioquímica, Química y Farmacia
Universidad Nacional de Tucumán
Tucumán, Argentina
and
Centro Científico y Tecnológico Tucumán
Tucumán, Argentina
Mario Eduardo Arena
Facultad de Bioquímica, Química y Farmacia
Universidad Nacional de Tucumán
Tucumán, Argentina
and
Centro Científico y Tecnológico Tucumán
Tucumán, Argentina
Francisco Noé Arroyo-López
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Azila Abdul Aziz
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
and
Faculty of Chemical Engineering
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
Ramlan Aziz
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
and
Faculty of Chemical Engineering
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
Fernanda Badotti
Departamento de Microbiologia
Universidade Federal de Minas Gerais
Belo Horizonte, Minas Gerais, Brazil
xiii
xiv
Jose María Baquero
Departamento de Biotecnologia de Alimentos
Instituto de la Grasa (CSIC)
Seville, Spain
Ágnes Baráth
Unit of Biology
Central Food Research Institute
Budapest, Hungary
Luis Roberto Batista
Departamento de Biologia
Universidade Federal de Lavras
Lavras, Minas Gerais, Brazil
Joaquín Bautista-Gallego
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Nabil Benomar
Departamento de Ciencias de la Salud
Universidad de Jaén
Jaén, Spain
Olinda Pérez Borla
Departamento de Ingeniería Química y en
Alimentos
Universidad Nacional de Mar del Plata
Mar del Plata, Argentina
and
Consejo Nacional de Investigaciones Científicas
y Técnicas
Buenos Aires, Argentina
Paola Carnevali
Bioprocesses and External Research
Barilla G. and R. F.lli SpA
Parma, Italy
Sam K. C. Chang
Department of Cereal and Food Sciences
North Dakota State University
Fargo, North Dakota
Ekachai Chukeatirote
School of Science
Mae Fah Luang University
Chiang Rai, Thailand
Contributors
Maurizio Ciani
Dipartimento di Scienze della Vita e
dell’Ambiente (DISVA)
Università Politecnica delle Marche
Ancona, Italy
Francesca Comitini
Dipartimento di Scienze della Vita e
dell’Ambiente (DISVA)
Università Politecnica delle Marche
Ancona, Italy
Aldo Corsetti
Department of Food Science
University of Teramo
Teramo, Italy
Katekan Dajanta
Faculty of Food and Agricultural Technology
Pibulsongkram Rajabhat University
Phitsanulok, Thailand
Lidia Dorantes-Alvarez
Graduados e Investigación en Alimentos
Escuela Nacional de Ciencias Biológicas-IPN
México City, México
Huseyin Erten
Department of Food Engineering
Cukurova University
Adana, Turkey
Adelfo Escalante
Departamento de Ingeniería Celular y Biocatálisis
Instituto de Biotecnología
Universidad Nacional Autónoma de México
Cuernavaca, Morelos, México
Lihua Fan
Agriculture and Agri-Food Canada
Atlantic Food and Horticulture Research Center
Kentville, Nova Scotia, Canada
Marcelo Fernández-Lahore
Downstream Processing Laboratory
Jacobs University
Bremen, Germany
Guadalupe Esquivel Flores
ILSI de México
Francisco Petrarca
México City, México
xv
Contributors
Antonio Gálvez
Departamento de Ciencias de la Salud
Universidad de Jaén
Jaén, Spain
Maria Gullo
Department of Agricultural Science
University of Modena and Reggio Emilia
Reggio Emilia, Italy
Pedro García-García
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Anna Halász
Unit of Biology
Central Food Research Institute
Budapest, Hungary
Antonio Garrido-Fernández
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Åse Solvej Hansen
Department of Food Science
University of Copenhagen
Frederiksberg, Denmark
Marisa S. Garro
Laboratorio de Tecnología y Desarrollo
Centro de Referencia para Lactobacilos
(CERELA-CONICET)
Tucumán, Argentina
Lisbeth Truelstrup Hansen
Department of Process Engineering and Applied
Science
Dalhousie University
Halifax, Nova Scotia, Canada
Martha Giles-Gómez
Departamento de Biología, Facultad de Química
Universidad Nacional Autónoma de México
México City, México
Humberto Hernández-Sánchez
Graduados e Investigación en Alimentos
Escuela Nacional de Ciencias Biológicas-IPN
México City, México
Paolo Giudici
Department of Agricultural Science
University of Modena and Reggio Emilia
Reggio Emilia, Italy
Y. H. Hui
Science Technology System
West Sacramento, California
Fátima C. O. Gomes
Departamento de Química
Centro Federal de Educação Tecnológica de
Minas Gerais
Belo Horizonte, Minas Gerais, Brazil
Silvia Nelina González
Facultad de Bioquímica, Química y Farmacia
Universidad Nacional de Tucumán
Tucumán, Argentina
and
Centro Científico y Tecnológico Tucumán
Tucumán, Argentina
and
Centro de Referencia para Lactobacilos
Tucumán, Argentina
Karendash González-Quijano
Graduados e Investigación en Alimentos
Escuela Nacional de Ciencias Biológicas-IPN
México City, México
M. Eugenia Jaramillo-Flores
Graduados e Investigación en Alimentos
Escuela Nacional de Ciencias Biológicas-IPN
México City, México
Rufino Jiménez-Díaz
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Vinod Kumar Joshi
Department of Food Science and Technology
Dr. Y. S. Parmar University of Horticulture and
Forestry
Solan, India
Suwimon Keeratipibul
Department of Food Technology
Chulalongkorn University
Bangkok, Thailand
xvi
Contributors
Catalina Elena Kotlar
Departamento de Ingeniería Química y en
Alimentos
Universidad Nacional de Mar del Plata
Mar del Plata, Argentina
and
Consejo Nacional de Investigaciones Científicas
y Técnicas
Buenos Aires, Argentina
Zhisheng Liu
Department of Cereal and Food Sciences
North Dakota State University
Fargo, North Dakota
Joseph Kreger
Department of Food Science and Human
Nutrition
University of Illinois
Urbana-Champaign, Illinois
Agustín López-Munguía
Departamento de Ingeniería Celular y Biocatálisis
Instituto de Biotecnología
Universidad Nacional Autónoma de México
Cuernavaca, Morelos, México
Patricia Lappe-Oliveras
Departamento de Botánica
Instituto de Bioligía
Universidad Nacional Autónoma de México
México City, México
Naphatrapi Luangsakul
Faculty of Agro-Industry
King Mongkut’s Institute of Technology Ladkrabang
Bangkok, Thailand
Soo-Yeun Lee
Department of Food Science and Human
Nutrition
University of Illinois
Urbana-Champaign, Illinois
Xinhong Liang
Department of Food Science
Henan Institute of Science and Technology
Xinxiang, Henan, China
Juliano De Dea Lindner
Bioprocess Engineering and Biotechnology
Department
Federal University of Paraná
Curitiba, Paraná, Brazil
and
Food Engineering Department
Santa Catarina State University
Pinhalzinho, Santa Catarina, Brazil
Antonio López-López
Departamento de Biotecnología de Alimentos
Universidad Nacional Autónoma de México
Cuernavaca, Morelos, México
Magdalena Martínez Cañamero
Departamento de Ciencias de la Salud
Universidad de Jaén
Jaén, Spain
Francisco Maugeri Filho
Food Engineering Department
State University of Campinas
Campinas, São Paulo, Brazil
Fabio Minervini
Dipartimento di Biologia e Chimica
Agroforestale ed Ambientale
Università degli Studi di Bari Aldo Moro
Bari, Italy
Haizhen Mo
Department of Food Science
Henan Institute of Science and Technology
Xinxiang, Henan, China
Hanifah Nuryani Lioe
Department of Food Science and Technology
Bogor Agricultural University
Bogor, Indonesia
Ljiljana Mojović
Department of Biochemical Engineering and
Biotechnology
University of Belgrade
Belgrade, Serbia
Shao Quan Liu
Department of Chemistry
National University of Singapore
Singapore
Olga Lucía Mondragón-Bernal
Food Science Department
Federal University of Lavras
Lavras, Minas Gerais, Brazil
xvii
Contributors
M. Victoria Moreno-Arribas
Instituto de Investigación en Ciencias de la
Alimentación (CIAL)
CSIC-UAM
Madrid, Spain
Rubén Moreno-Terrazas
Departamento de Ingeniería y Ciencias Químicas
Universidad Iberoamericana
México City, México
Erika Neely
Department of Food Science and Human
Nutrition
University of Illinois
Urbana-Champaign, Illinois
Scot Nelson
Department of Plant and Environmental
Protection Sciences
University of Hawaii at Manoa
Honolulu, Hawaii
Alessandro Nogueira
Departamento de Engenharia de Alimentos
Universidade Estadual de Ponta Grossa
Ponta Grossa, Paraná, Brazil
Norhayati Mohamed Noor
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
Norfarahiyah Aman Nor
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
María Llanos Palop
Department of Analytical Chemistry and Food
Technology
University of Castilla–La Mancha
Toledo, Spain
Lidia Parada-Dorantes
Universidad del Caribe
Cancún, Quintana Roo, México
Ana Lúcia Fernandes Pereira
Departamento de Tecnologia de Alimentos
Universidade Federal do Ceará
Fortaleza, Ceará, Brazil
Alejandra Graciela Ponce
Departamento de Ingeniería Química y en
Alimentos
Universidad Nacional de Mar del Plata
Mar del Plata, Argentina
and
Consejo Nacional de Investigaciones Científicas
y Técnicas
Buenos Aires, Argentina
M. Ángeles Pozo-Bayón
Instituto de Investigación en Ciencias de la
Alimentación (CIAL)
CSIC-UAM
Madrid, Spain
Rubén Pérez Pulido
Departamento de Ciencias de la Salud
Universidad de Jaén
Jaén, Spain
Marica Rakin
Department of Biochemical Engineering and
Biotechnology
University of Belgrade
Belgrade, Serbia
Lebaka Veeranjenaya Reddy
Department of Microbiology
Yogi Vemana University
Kadapa, India
Obulam Vijaya Sarathi Reddy
Department of Microbiology
Sri Venkateswara University
Tirupati, India
Maria Isabel Rodrigues
Food Engineering Department
State University of Campinas
Campinas, São Paulo, Brazil
Sueli Rodrigues
Departamento de Tecnologia de Alimentos
Universidade Federal do Ceará
Fortaleza, Ceará, Brazil
Juan José Rodríguez-Bencomo
Instituto de Investigación en Ciencias de la
Alimentación (CIAL)
CSIC-UAM
Madrid, Spain
xviii
Francisco Rodríguez-Gómez
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Verónica Romero-Gil
Departamento de Biotecnología de Alimentos
Instituto de la Grasa, CSIC
Seville, Spain
Carlos A. Rosa
Departamento de Microbiologia
Universidade Federal de Minas Gerais
Belo Horizonte, Minas Gerais, Brazil
Sara I. Roura
Departamento de Ingeniería Química y en
Alimentos
Universidad Nacional de Mar del Plata
Mar del Plata, Argentina
and
Consejo Nacional de Investigaciones Científicas
y Técnicas
Buenos Aires, Argentina
Muhammad Roji Sarmidi
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
and
Faculty of Chemical Engineering
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
Graciela Savoy de Giori
Laboratorio de Tecnología y Desarrollo
Centro de Referencia para Lactobacilos
(CERELA-CONICET)
Tucumán, Argentina
and
Facultad de Bioquímica, Química y Farmacia
Universidad Nacional de Tucumán
Tucumán, Argentina
Rosane Freitas Schwan
Departamento de Biologia
Universidade Federal de Lavras
Lavras, Minas Gerais, Brazil
Contributors
Maja Vukašinović Sekulić
Department of Biochemical Engineering and
Biotechnology
University of Belgrade
Belgrade, Serbia
Susana Seseña
Department of Analytical Chemistry and Food
Technology
University of Castilla–La Mancha
Toledo, Spain
Nur Arbainah Shamsul Annuar
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
Rakesh Sharma
Department of Food Science and Technology
Dr. Y. S. Parmar University of Horticulture and
Forestry
Solan, India
Cristina Ferreira Silva
Departamento de Biologia
Universidade Federal de Lavras
Lavras, Minas Gerais, Brazil
Carlos Ricardo Soccol
Bioprocess Engineering and Biotechnology
Department
Federal University of Paraná
Curitiba, Paraná, Brazil
Vanete Thomaz Soccol
Bioprocess Engineering and Biotechnology
Department
Federal University of Paraná
Curitiba, Paraná, Brazil
Lisa Solieri
Department of Agricultural Science
University of Modena and Reggio Emilia
Reggio Emilia, Italy
Michele Rigon Spier
Bioprocess Engineering and Biotechnology
Department
Federal University of Paraná
Curitiba, Paraná, Brazil
xix
Contributors
Marzia Stringini
Dipartimento di Scienze della Vita e
dell’Ambiente (DISVA)
Università Politecnica delle Marche
Ancona, Italy
Caroline Tiemi Yamaguishi
Bioprocess Engineering and Biotechnology
Department
Federal University of Paraná
Curitiba, Paraná, Brazil
Chua Lee Suan
Institute of Bioproduct Development
Universiti Teknologi Malaysia
Skudai, Johor, Malaysia
Yong Yang
Institute of Food Safety
China Jiliang University
Hangzhou, China
Jyoti Prakash Tamang
Department of Microbiology
Sikkim University
Sikkim, India
Masaaki Yasuda
Department of Bioscience and Biotechnology
University of the Ryukyus
Okinawa, Japan
Hasan Tanguler
Department of Food Engineering
Cumhuriyet University
Kampus/Sivas, Turkey
Sirma Yegin
Department of Food Engineering
Ege University
Izmir, Turkey
and
Downstream Processing Laboratory
Jacobs University
Bremen, Germany
Nantawan Therdthai
Faculty of Agro-Industry
Kasetsart University
Bangkok, Thailand
Luciana Porto de Souza Vandenberghe
Bioprocess Engineering and Biotechnology
Department
Federal University of Paraná
Curitiba, Paraná, Brazil
Gilvan Wosiacki
Departamento de Engenharia de Alimentos
Universidade Estadual de Ponta Grossa
Ponta Grossa, Paraná, Brazil
Zsolt Zalán
Unit of Biology
Central Food Research Institute
Budapest, Hungary
Weibiao Zhou
Department of Chemistry
National University of Singapore
Singapore
43
Pulque Fermentation
Adelfo Escalante, Martha Giles-Gómez, Guadalupe Esquivel Flores, Violeta Matus Acuña,
Rubén Moreno-Terrazas, Agustín López-Munguía, and Patricia Lappe-Oliveras
CONTENTS
43.1 Introduction................................................................................................................................... 691
43.2 History of Pulque.......................................................................................................................... 692
43.2.1 Origins and Pre-Hispanic Production.............................................................................. 692
43.2.2 Pulque Production during the Spaniard Colonial Period................................................. 692
43.2.3 Production during the 19th and 20th Centuries............................................................... 693
43.2.4 Situation in the Early 21st Century.................................................................................. 694
43.3 Agave Species Used for Aguamiel Extraction.............................................................................. 694
43.3.1 Aguamiel Composition and Properties............................................................................ 695
43.3.1.1 Physicochemical Characteristics of Aguamiel................................................. 695
43.4 Pulque Production......................................................................................................................... 695
43.4.1 Traditional Pulque Production......................................................................................... 695
43.5 Microbiology and Biochemistry of the Fermentation................................................................... 697
43.5.1 Microbial Diversity in Pulque: Defining an Essential Microbiota?................................. 697
43.5.2 Microbial Dynamics of the Fermentation........................................................................ 698
43.5.3 Physicochemical Changes during Pulque Fermentation.................................................. 699
43.6 Pulque: Nutritional and Traditional Medicine Aspects................................................................ 699
43.6.1 Pulque as Probiotic and Prebiotic Agent.......................................................................... 700
43.7 Pulque Industrialization and Major Technological Challenges.....................................................701
43.7.1 Pulque Fermentation Medium Composition.....................................................................701
43.7.2 Fermentation..................................................................................................................... 702
43.7.3 Pulque Flavor.................................................................................................................... 702
43.7.4 Pulque Conservation......................................................................................................... 703
References............................................................................................................................................... 704
43.1 Introduction
Pulque is probably the oldest and most traditional Mexican alcoholic beverage, prepared, and consumed
since pre-Hispanic times. Due to its great historical, religious, social, medical, and economical importance, it is the most widely studied beverage from the anthropological and scientific points of view
(Loyola-Montemayor 1956; Godoy et al. 2003).
It is a milky white, viscous, slightly acidic alcoholic beverage traditionally produced by spontaneous
fermentation of aguamiel, the sugary sap extracted from several Agave species that grow in the central Mexican plateau states (Loyola Montemayor 1956; Steinkraus 1996; García-Mendoza 1995, 1998;
Blomberg 2000; Godoy et al. 2003). In pre-Hispanic times, pulque played an important role in the religion, rituals, divination, and curing of the indigenous communities (Bruman 2000). Currently, pulque
persists as a typically popular beverage, and it is sold in pulquerías as well as in some restaurants. It is
also produced at a small industrial scale using a mixed starter culture and controlling the quality and
safety of the product, which is canned and exported (Ramírez et al. 2004).
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Handbook of Plant-Based Fermented Food and Beverage Technology
In this chapter, a broad view of pulque is covered: a historical review is presented, from its discovery and relevance during the pre-Hispanic period, its role during colonial times, its consolidation, and production from the 15th to the 20th centuries, until the recent scientific and technological
efforts to reincorporate pulque to the Mexican traditional cuisine. The different Agave species used
in the production of pulque are mentioned as well as their actual distribution and density in the agave
plantations. The traditional and industrial pulque production processes are described, including the
microbiota involved as well as the biochemical changes that take place both in the spontaneous and the
controlled fermentation. To understand the relevance and potential benefits of pulque among consumers, its main chemical and nutritional properties, including its health benefits, are discussed. Finally,
the chapter concludes with a description of the actual status of pulque in Mexican society as well as
the social, scientific, and technological challenges that have to be faced in order to achieve the rescue
of this ancestral beverage.
43.2 History of Pulque
43.2.1 Origins and Pre-Hispanic Production
Pulque represents a Mexican icon and is probably the oldest and most traditional Mexican alcoholic beverage, as attested by archaeological vestiges. The ancient Aztecs called it metoctli or agave wine (from
the nahuatl language metl = agave or maguey + octli = wine) as well as iztacoctli or white wine (from
izac = white + octli = wine): they referred to the spoiled beverage with unpleasant odor and flavor as
poliuhquioctli (from poliuhqui = spoiled or rotted + octli = wine), so it is probably from this last term that
the word pulque was adopted by the Spaniards to designate the freshly produced beverage (Robelo 1948).
It is highly probable that ancient Otomíes were the first to prepare pulque toward the year 2000 BC,
inheriting the elaboration process to almost all cultures of the central Mexican plateau (Martín del
Campo 1938; Blomberg 2000). However, different legends attribute its origin to the Tolteca civilization
(1021–990 BC) in the city of Tula. One of these legends considers that maguey and pulque are both a
gift of the goddess Mayahuel (Gonçalves de Lima 1978; Ramírez Castañeda 1994). There is consensus
among historians based on archeological evidences (200 BC–300 AD) and information from pre- and
post-Hispanic codices from several important Mesoamerican cultures that pulque had a dominating
presence in their daily life and a primordial role in religious and war rituals (Diguet 1928; Martin del
Campo 1938; Sánchez-Marroquín 1949; Gonçalves de Lima 1978; Vallejo 1992).
Aztec civilization developed a full dominion of aguamiel and pulque production techniques incorporating to their diet its nutritional benefits and healing properties. This civilization considered pulque
a divine food, a medicine, and a ritual element that allowed a closer relation with their cosmology. As
the dominant culture in the Mexican plateau, the Aztecs established the moral guidelines for pulque
consumption and religious use: only the children and youths, the elder, pregnant or nursing women to
increase milk production, priests and warriors, or anyone before sacrifice were allowed to consume it,
but excessive consumption was harshly punished, in some cases including the death penalty (LoyolaMontemayor 1956; Corcuera de Mancera 1991; Godoy et al. 2003). Aztecs’ conquered cultures were
forced to deliver aguamiel and pulque as tribute (Gonçalves de Lima 1978, 1990; Soberón-Mora 1998).
43.2.2 Pulque Production during the Spaniard Colonial Period
When the Aztec Empire fell, pulque lost its religious importance prevailing as a traditional popular
beverage until today. At the beginning of the colonial period, the Spaniards promoted its consumption among the indigenous groups and recent immigrant slaves as a mean to control them (Corcuera
de Mancera 1991; Ramirez et al. 2004). The fast growth of the productive and commercial activities
introduced by the Spaniards during the 17th century gradually incorporated the production and commercialization of pulque. These activities flourished at the end of the 18th century with the development
of the haciendas pulqueras (large farms producing pulque) devoted to the cultivation of Agave plants,
extraction of aguamiel, and production of pulque, which was transported mainly to the cities of Mexico
Pulque Fermentation
693
and Puebla. These production organizations prospered in the actual central Mexican plateau states of
Hidalgo (mainly in the Apan prairies), Tlaxcala, Puebla, Querétaro, Morelos, and Michoacán.
Since 1779, pulque production and commerce was strongly regulated, and special privileges were
granted to Spaniards who controlled these agroindustry (Loyola-Montemayor 1956), becoming one of
the most important economic activities in the New Spain. Taxes imposed to pulque became the fifth biggest income of the Real Hacienda and the Spanish Crown received from the pulque industry 4.3 million
pesos in 1762 and 32 million from 1763 to 1809 (Segura 1901).
43.2.3 Production during the 19th and 20th Centuries
During the independence movement (1810–1821), pulque production suffered a drastic reduction. Once
México became independent, government regulations regarding pulque production, quality, commerce,
and taxes consolidated again the haciendas pulqueras, allowing them to cope with the increasing demand
of the beverage by the rapidly growing population (Loyola Montemayor 1956). An important factor
in this process was the introduction in 1867 of the railway system, which in particular connected the
Apan prairies with the main consumption centers. In this context, the pulque agroindustry flourished and
reached its best times at the beginning of the 20th century with an annual production of 500 million L
obtained from around 140 million agave plants. In 1905, 350,000 L of pulque were consumed only in
Mexico City. This agroindustry controlled the agave plantations and the production and retail sale of
pulque, activities that originated great fortunes and the emergence of a new social economic class called
the “pulque aristocracy” (Ramírez-Rancaño 2000; CECULTAH 2010).
In 1909, the owners of more than 50 haciendas created the Compañía Expendedora de Pulques, with
the purpose of controlling production, distribution, and sale of pulque in Mexico City and to expand
the hacienda activities to the elaboration, distribution and commerce of other agave products, including
aguamiel syrups, glues, pulque distillates, and medical products, increasing the benefits of the agave
industry (Gobierno del Estado de Hidalgo: Museo Nacional de Culturas Populares 1988).
By 1910, the pulque industry was affected again by the revolutionary movement (Soberón-Mora 1998).
The social instability together with the introduction of the brewing companies and a strong campaign
against pulque had a negative impact in this agroindustry. By 1914, haciendas pulqueras were seized and
land reform started fractionating the agricultural fields. The elimination of haciendas pulqueras became
a fact with the promulgation of a new constitution in 1917, which established in the 27th article the agricultural reform, which promoted the return and redistribution of the land to the peasants; this was the
formal end of the three centuries of splendor of most of the haciendas pulqueras (Loyola Montemayor
1956; Ramírez-Rancaño 2000). Despite the land redistribution, the abundance of agave cultivars allowed
the development of a new pulque industry that was able to cope with the decreased beverage demand.
Therefore and until the 1920s, fresh pulque produced in several regions of the country was delivered
daily by train to Mexico City. During 1934–1940, antialcoholic campaigns were established, particularly
against pulque, as it was considered dirty and unhealthy, due to the lack of control in the production
process and adulteration during its distribution after certification through the former pulque customs of
Pantaco, Cuautepec, and Ticoman, where rigorous norms were applied until the seventies to authorize
the entrance of pulque barrels to Mexico City (Gobierno del Estado de Hidalgo: Museo Nacional de
Culturas Populares 1988). However, during the following decades the already mentioned problems, as
well as the excessive and irrational exploitation of the agave plantations and the introduction of beer
as an alternative beverage, considerably deteriorated the pulque industry (Loyola-Montemayor 1956;
Ramírez-Rancaño 2000).
It was not until 1960 that considerable efforts were made in order to recover this important national
heritage with the creation of the Patronato del Maguey, with the aim to improve the culture and exploitation of Agave through modifications in both the production and the distribution process. Canning was
introduced (350 mL cans and commercialized under the registered trade name Magueyin) as an alternative way of distribution, efforts were made to improve the hygienic conditions of the beverage, and new
uses of the Agave plants were developed to increase the income of the peasants (Gobierno del Estado
de Hidalgo: Museo Nacional de Culturas Populares 1988; Steinkraus 1996). Unfortunately, canned pulque was not accepted by the traditional consumer as the product lacked the distinguishing sensorial
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Handbook of Plant-Based Fermented Food and Beverage Technology
properties of fresh pulque and had a higher price. As a consequence, this industrialization attempt failed
as well as subsequent efforts to commercialize other former patented pulque brands.
In the 20th century, according to the agronomical census, from 1998 to 2002, the surface devoted to
Agave pulquero was reduced with the consequent decrease in pulque production. Nowadays, the most
important agave pulquero production areas are located in the central states of Hidalgo, Tlaxcala, and
Estado de México, and in some regions of Querétaro, Puebla, Morelos, and Michoacán. By the end of the
last century, the depressed pulque industry situation was the consequence of the loss of cultivars, the lack
of an efficient technology, and the low demand by consumers (Loyola-Montemayor 1956; Gobierno del
Estado de Hidalgo: Museo Nacional de Culturas Populares 1988; Ramírez-Rancaño 2000).
43.2.4 Situation in the Early 21st Century
Currently, pulque is considered a traditional popular beverage that practically has disappeared from the
urban market, only found in pulquerías located in popular districts of Mexico City. In the rural areas,
near the production regions, pulque is still the preferred stimulating beverage of the low income class.
It is consumed as part of their daily diet and is part of the tradition in all sorts of festivities, from births
and baptisms to weddings and funerals. It is also common to find production for self-consumption among
agave owners. Important efforts are being made to fight against its disappearance: it is being promoted in
Mexican food restaurants and gastronomic festivals, while traditional pulquerías are recovered. In these
traditional places pulque is now exclusively prepared with high quality raw materials and under hygienic
conditions (Poblet 1995).
Nevertheless, pulque production remains as a predominantly traditional and small-scale industry that
has not been able to massively reach local (other than Mexico City) or international markets (Ramírez
et al. 2004). However, Nectar del Razo, in Tlaxcala, and Desarrollo Agropecuario del Altiplano, in
Puebla, export their novel canned pulque product to different countries all around the world. In both
companies, the traditional elaboration process has been modified to obtain a stable product with a long,
shelf-life, which is sold white, that is to say, plain or added with fruits.
43.3 Agave Species Used for Aguamiel Extraction
Pulque is produced from aguamiel extracted from native Agave species, mainly A. salmiana var. salmiana (green or meek agave), A. atrovirens (white agave), and A. mapisaga (Mexican or long hand agave),
and in lower proportion from A. lehmannii and A. altisima (Loyola-Montemayor 1956; García-Mendoza
1995, 1998), mostly distributed in the states of the central Mexican plateau where pulque has been mainly
produced and consumed. These species grow in a vast region of arid, semiarid, and temperate zones,
in sandy, poor, well-drained soils, with scare, and irregular precipitations. Preventing soil erosion, they
can be found as single crop or protecting or limiting homes or farm lands (Loyola-Montemayor 1956;
Steinkraus 1996; Ramírez et al. 2004; Ortiz-Basurto et al. 2008; Lappe-Oliveras et al. 2008).
Agave is a plant relatively easy to cultivate. Propagation may be carried out by transplanting young vigorous offsprings from adult plants, called matecuates. An adult plant can produce above 50 matecuates
that grow around the mature plant. Young offsprings are harvested in the rainy season and transplanted
into a new magueyera or agave plantation for aguamiel production where the young plants are arranged
on parallel long rows called melgas or metepnatle (maguey wall), separated by 2–5 m. Alternatively,
agave seeds can be grown in seedbeds under controlled conditions (Monterrubio 2007).
Aguamiel is extracted from 7- to 10-year-old mature agaves that are about to produce their inflorescence (quiote). Mature plants are castrated or broken by eliminating the floral bud, leaving a cavity (cajete) in the center of the agave stem. This operation is usually performed in early spring or late
autumn. If the plant inflorescence grows, maguey will never produce aguamiel, and if castration is
performed before agave maturation, the yield in aguamiel production will be scare. The resulting cavity
is covered with agave leaves or with a sized stone to avoid the exposition of the cavity to the environment
(Loyola-Montemayor 1956; Gobierno del Estado de Hidalgo: Museo Nacional de Culturas Populares
1988). An aging period follows castration to allow the maturation of the central leaves and the increase
Pulque Fermentation
695
of sap sugar content: 7%–14% w/v, depending of the agave species (Lappe-Oliveras et al. 2008). The
healed cavity is scraped to open the vessels, promoting the sap flow and its accumulation in the cavity.
The sap is collected twice a day (at daybreak and dusk) by oral suction through a dried gourd (Lagenaria
siceraria) called acocote. Production of aguamiel lasts from 3 to 6 months until the plant dies, with an
approximate production of 1000 L/plant (Loyola-Montemayor 1956; Gobierno del Estado de Hidalgo:
Museo Nacional de Culturas Populares 1988; Steinkraus 1996). The volume of aguamiel produced varies
throughout the harvesting period; at the beginning the plant produces around 0.4 L/plant/day, an amount
that increases to 4 or 6 L/day during the next 3–6 months and then decreases to 0.4 L before the plant dies
(Ortiz-Basurto et al. 2008). Once the plant dies, it is removed and substituted by an offspring, as already
described (Monterrubio 2007; Lappe-Oliveras et al. 2008).
43.3.1 Aguamiel Composition and Properties
43.3.1.1 Physicochemical Characteristics of Aguamiel
Aguamiel is a milky white, lightly cloudy, thick, sweet, fresh-flavored, and lightly acid agave sap. It
contains water, sugars, proteins, gums, and mineral salts as the most important components (Ramírez et
al. 2004; Ortiz-Basurto et al. 2008; Escalante et al. 2008). The Mexican regulation NMX-V-022 (Banco
de Normas Mexicanas 2010a) defines two types of aguamiel for pulque production. Type I refers to a
best quality aguamiel (cleanest and with a highest sugar content, pH < 6.6–7.5 and < 0.9–1.03 mg lactic
acid/100 mL) while type II includes slight acid aguamiel (pH < 4.5 and < 4 mg lactic acid/100 mL).
According to several sources, the main compounds present in the sap are 0.3 to 0.4 g/100 mL of all
essential amino acids with exception of methionine, vitamins (mg/100 mL, 0.02 riboflavin, 0.06 thiamine, 0.45 niacin, 19.60 biotin, 21.64 p-aminobenzoic acid, 65.17 panthothenic acid, 22.98 pyridoxine,
and 9.0 ascorbic acid), sugars (glucose, sucrose, fructose, fructo-oligosaccharides, and inulin), minerals
(mg/100 mL, 25 sodium, 9 magnesium, 1.1 iron, 0.8 zinc, 0.3 copper, and 0.3 manganese). Aguamiel
constitutes a water substitute or an alternative food in places where water is scare or of bad quality or
where protein is not available (Sánchez-Marroquín and Hope 1953; Ortiz-Basurto et al. 2008; GarcíaGaribay and López-Munguía 1993; Steinkraus 1996; Lappe-Oliveras et al. 2008).
The amount and composition of aguamiel during the production period varies depending on the Agave
species, the cultivation conditions of the plant, the season of the year, the air relative humidity, and the
soil properties (Sánchez-Marroquín 1970). However, in the physicochemical analysis of aguamiel from
A. mapisaga harvested during 5 months, Ortiz-Basurto et al. (2008) found no significant changes in
titrable acidity, pH (above 4.5), and dry matter content, but a slight decrement in fructose content was
observed in samples collected after 3.5 months, concluding that the quality of aguamiel remains constant
during all the harvesting period.
43.4 Pulque Production
The Mexican regulation NMX-V-037 (Banco de Normas Mexicanas 2010b) recognizes two types of
pulque: type I includes pulque de semilla or pulque pie de cuba (seed or starter) and type II refers to commercial pulque. Pulque starter is prepared to increase the natural microbiota that determines the correct
fermentation in the elaboration of the beverage in a tank exclusively devoted for this purpose. Type I sap
is inoculated with pulque starter, and is used to establish the optimum biochemical equilibrium between
the fermentable substrates and the microorganisms involved in fermentation; it is the base of commercial
production (Loyola-Montemayor 1956; Ramírez et al. 2004; Lappe-Oliveras et al. 2008).
43.4.1 Traditional Pulque Production
The traditional process of pulque production has remained relatively unchanged. The process starts
with the daily collection of aguamiel and its immediately transference to a container, traditionally a
pork or goat skin bag called cuero (Figure 43.1 shows several stages and utensils used during aguamiel
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(a)
(d)
(b)
(e)
(c)
(f )
FIGURE 43.1 Aguamiel extraction, transport, and pulque fermentation during the early 20th century (a–c) and at present (d–e, from the aguamiel and pulque production location Las Mesas, Tlaxcala state). (a) Aguamiel extraction by the
tlachiquero with an acocote, cajete scrapper tool (right hand), and skin bag to transport aguamiel to fermentation vats;
(b) aguamiel and pulque transportation in skins bags and barrels; (c) former vats made with leather in a large tinacal;
(d) modern tlachiquero with an acocote and scrapper tool; (e) aguamiel and pulque transport to small rural pulquerías;
(f) large plastic barrels (200 L) in a small tinacal. (a–c, Reproduced with permission of Sistema Nacional de Fototecas,
México. a, ©609476, Conaculta, INAH, Sinafo, FN, México; b, ©611009, Conaculta, INAH, Sinafo, FN, México; c, ©6535,
Conaculta, INAH, Sinafo, FN, México.)
extraction and pulque production) or a wood container known as castaña; recently, plastic containers
have been introduced. After sap collection, the cajete walls are scraped again to maintain the vessels
open and a constant aguamiel flow.
Fermentation is carried out for several hours, depending on aguamiel quality, pulque starter maturity,
the sap, and seed microbiota, and the season (environmental temperature changes). The degree of fermentation is adequate when a specific alcoholic degree and viscosity are reached (Loyola-Montemayor
1956; Steinkraus 1996).
Pulque fermentation is performed in containers made from different materials (leather, wood, plastic,
glass fiber) placed in a closed room (tinacal, with small windows to control the temperature), as in the
Pulque Fermentation
697
haciendas pulqueras of the Spaniard colonial period. In pre-Hispanic times pulque fermentation was
carried out in clay pots; during the colonial period these kind of containers were replaced by leather reservoirs that were used until 1900, when wood vats with higher capacities (800–1000 L) were introduced.
In most tinacales, the fermentation tanks are generally arranged in parallel, while in some others they
are placed around a circle (Bellingeri Martini 1980).
To produce the starter, a small amount of aguamiel (10–15 L) preferably with an adequate density
and taste is poured into a devoted vat (located in a small room next to the tinacal) and spontaneously
fermented until alcoholic and acetic tastes are detected or until a white layer called zurron is formed on
the surface. This process is performed at room temperature and lasts several weeks depending on the
season: 1 week in summer, while 3 or 4 weeks in winter (Loyola-Montemayor 1959; Bellingeri Martini
1980; Gobierno del Estado de Hidalgo: Museo Nacional de Culturas Populares 1988; Steinkraus 1996;
Lappe-Oliveras et al. 2008).
Small amounts of fresh aguamiel are constantly poured until the vat is filled; this amount of seed
should be enough to sustain commercial pulque production. Afterward, the starter is distributed in several vats, a process known as tirar puntas, and the fermentation is started with the addition of fresh aguamiel to the vats. An empirical rule applied to perform a good fermentation is to mix more starter than
aguamiel during the cold season and less starter than aguamiel during the hot season. The fermentation
finishes after 6 or 14 hours depending on seed and aguamiel quality, microbial content, and environmental conditions. As all these parameters affect the fermentation process it is not strange to find pulque
with a wide variety of properties and compositions (Loyola-Montemayor 1956; Raminez et al. 2004).
According to its quality, two types of pulque are recognized: fine or best-quality pulque obtained by
fermentation of high-quality aguamiel from A. salmiana var. salmiana, A. mapisaga, and A. atrovirens,
grown in cold weather and dried land, and poor quality pulque or tlachique produced by fermentation
of poor quality aguamiel (low sugar content) from agaves grown in humid lands and temperate weather
(Loyola-Montemayor 1956).
Due to the number and distribution of places where traditional pulque is produced, it is difficult to
have a strict regulation and to avoid adulteration of the product, which is sometimes diluted with water
to increase the volume of product, with gums to mask a deficient fermentation or with sugars to avoid
offending flavors (Monterrubio 2007). Pulque is consumed alone or mixed with single or combined
macerated fruits (strawberry, guava, maracuya, figs), vegetables (alfalfa, celery, tomatoes), nuts (pecans,
almonds, peanuts, pine nuts, coconut), or condiments (cinnamon, black, or red pepper, coffee), beverages
named curados.
43.5 Microbiology and Biochemistry of the Fermentation
43.5.1 Microbial Diversity in Pulque: Defining an Essential Microbiota?
Aguamiel constitutes a favorable medium for the proliferation of numerous microorganisms associated
to the environment: from the cajete walls, incorporated with the scraping or extraction tools, from dust,
or from insects such as Drosophila spp. This microbiota complements that of the starter present in the
fermentation container to perform a series of distinctive fermentations processes that characterize pulque: acid (lactic and acetic), alcoholic (production of ethanol), and viscous (production of extracellular
polysaccharides, EPS) (Sánchez-Marroquín and Hope 1953; Loyola-Montemayor 1956; Gonçalves de
Lima 1990; García-Garibay and López Munguía 1993).
Early microbiological studies performed by Sánchez-Marroquín during 1946–1957, resulted in a
description of the pulque microbiota and the proposal of Lactobacillus spp., Leuconostoc mesentero­
ides, Zymomonas mobilis, and the yeast Saccharomyces cerevisiae as the essential microorganisms in
pulque fermentation. Using purified strains of these organisms, a mixed inoculum was developed and
applied in controlled fermentation processes using sterilized aguamiel, demonstrating that the final fermented product was comparable in alcohol content, viscosity, and acidity to the traditional high-quality
pulque produced commercially (Sánchez-Marroquín and Hope 1953; Sánchez-Marroquín et al. 1957).
Although these results suggest the essential role of the selected microorganisms, recent studies on the
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microbiology of aguamiel and pulque demonstrate the presence of a complex microbial diversity of bacteria and yeast (Escalante et al. 2004, 2008; Lappe-Oliveras et al. 2008).
Analysis of the bacterial diversity in pulque samples collected from Estado de México, Morelos, and
Hidalgo demonstrate the presence of common species in all analyzed samples, but also unique species
in pulque from each of the three regions. Lactic acid bacteria (LAB) related to L. acidophilus (homofermentative) was the most abundant group present in the samples, while the previously reported L. mesenteroides and Z. mobilis (considered as essential microorganisms) were detected in a minor proportion
only in two of the studied samples (Escalante et al. 2004).
43.5.2 Microbial Dynamics of the Fermentation
Escalante et al. (2008) found in aguamiel an initial total count of 1.3 × 107 CFU/mL mesophilic bacteria, 3.2 × 108 CFU/mL LAB and 3.1 × 104 CFU/mL yeasts; in a sample taken after addition of the fresh
aguamiel to the seed (corresponding to the initial fermentation time: T0) the total yeast count increased
to 8.8 × 106 CFU/mL, while aerobic mesophilic bacteria and LAB counts were not modified: 1.2 × 107
and 1.5 × 108 CFU/mL, respectively. After 3 hours of fermentation, yeast increased to 1.4 × 107 CFU/mL,
maintaining this level until the end of fermentation (6 hours); total bacteria counts remained constant
(Escalante et al. 2008).
Bacterial diversity detected in aguamiel was composed mainly by L. mesenteroides, L. citreum, L.
kimchi, and the Proteobacteria Acinetobacter radioresistens, Erwinia rhapontici, and Enterobacter spp.
At T0, they were detected mainly some species observed in aguamiel and others present in the seed:
L. mesenteroides, L. citreum, Lactobacillus spp. (related to L. acidophilus), and the Proteobacteria E.
agglomerans (both in high proportion), Kluyvera ascorbata, Serratia grimensis, and the acetic acid bacteria (AAB) Acetobacter malorum. As fermentation proceed the population of L. kimchi, one of the most
abundant LAB detected in aguamiel, decreased; those of L. mesenteroides and L. citreum remained relatively constant, while populations of Lactococcus lactis and Z. mobilis were detected in low proportion.
After 3 hours of fermentation, important changes in bacterial diversity were observed: Lactobacillus
spp., L. mesenteroides, E. agglomerans, and A. malorum became the dominant species; L. acidophilus
and L. citreum were detected in low proportion; L. lactis, Z. mobilis, and A. radioresistens remained
relatively constant compared to T0, whereas S. grimensis and K. ascorbata were no longer found. At
the end of the fermentation (6 hours), the bacteria diversity was mainly composed by the homofermentative L. acidophilus and Lactobacillus spp., the heterofermentative L. mesenteroides, Lactococcus lactis
subsp. lactis, and A. malorum.
These results demonstrate that although no changes occurred in total bacteria counts during a 6-hour
fermentation, important changes take place in species distribution.
Lactobacillus species have been previously detected in aguamiel and pulque fermentation, but no
detailed information concerning the identity and role of this LAB group is available. Sánchez-Marroquín
and Hope (1953) reported that lactobacilli are responsible for the assimilation of 53% of the sugars present in aguamiel. L. acidophilus, L. hilgardii, and the heterofermentative L. sanfranciscensis are capable
to produce dl-lactic acid and to ferment glucose and fructose present in aguamiel, but only the former is
able to ferment sucrose (Carr et al. 2002).
Lactobacilli are naturally associated to plants, fermented vegetables, fermented doughs, and alcoholic
distilled beverages such as malt whiskey (van Beek and Priest 2000). The ability of LAB to grow in acid
environments and tolerate high ethanol concentrations could explain their high abundance during pulque
fermentation (final pH = 4.5 and 4%–6% ethanol, Escalante et al. 2008). The presence of the AAB A.
pomorum, A. malorum, and Gluconobacter oxydans in different pulque samples and the high production
of acetic acid in comparison to lactic acid after 6 hours of fermentation (Escalante et al. 2004, 2008),
suggests an important role of these bacteria in pulque fermentation.
Yeast identified along pulque fermentation belong to Saccharomyces (S. cerevisiae, S. bayanus, S.
paradoxus) and non-Saccharomyces (Candida spp., C. parapsilosis, C. lusitaniae, Kluyveromyces
marxianus, K. lactis, Hanseniaspora uvarum, Pichia spp., P. guilliermondii, Torulaspora delbrueckii)
are capable to produce ethanol from glucose, fructose, and sucrose and to synthesize nutritive amino
acids and vitamins as well as flavor-volatile compounds that influence the quality and the aromatic
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699
profile of the beverage. At early fermentation stages, the non-Saccharomyces yeasts develops, but as
ethanol concentration increases, they are replaced by primary ethanol-producing yeasts such as S. cerevisiae and several K. marxianus ethanol-tolerant strains. Both yeasts species predominate throughout the
fermentation until 6% of ethanol is reached (Lappe-Oliveras et al. 2008).
43.5.3 Physicochemical Changes during Pulque Fermentation
As already described, aguamiel is a sugar-rich sap containing sucrose, fructose, glucose, and fructo-​
oligosaccharides with a relatively acid pH (7.5–4.5), whereas pulque is an acidic (pH 4.3–3.5, total
acidity of 0.75–0.4 g lactic acid/100 mL of pulque), alcoholic (4%–6%), and viscous beverage (Banco
de Normas Mexicanas 2010b). Fresh aguamiel used to start fermentation with an initial pH of 6.0,
decreases to 4.5 after mixing with the seed, and to 4.1 at the end of the process after 6 hours of
fermentation. These environmental conditions favor the appearance of an acid- and ethanol-tolerant
microbiota toward the end of the fermentation and probably affect the survival of non–acidic-resistant
bacteria and yeast species that apparently disappear during the process as did the possible antimicrobial
activity of LAB (production of bacteriocins and H2O2) and Z. mobilis subsp. mobilis (production of
acetylmetylcarbinol) (Escalante et al. 2008; Lappe-Oliveras et al. 2008).
Fresh aguamiel has a mild temperature (17°C–20°C), which increases several degrees after inoculation
with the seed as a consequence of the microbial activity. In a 6-hour fermentation carried out to assess
the overall metabolic activity of the microbial community in aguamiel and pulque samples from the state
of Morelos, the concentrations of sucrose, glucose, fructose, and fermentation products were determined
during the process. Total sugars consumed after 6 hours of fermentation were 228.4 mM hexose equivalents (equivalent to 1370.5 mM of carbon, which was found as CO2). It was assumed, in molar terms, that
the amount of CO2 produced is equal to the amount of acetic acid and ethanol. If 176.5 mM of carbon
corresponding to 5.3 g/L of carbohydrates in EPS are also included, then 66.2% of total C was recovered
in products. Ethanol reached a final value of 142.17 mM carbon equivalents, while 35.73 and 6.32 mM,
respectively, of acetic and lactic acid concentrations were reached (Escalante et al. 2008).
Viscosity is one of the distinctive properties of pulque. This characteristic has been associated with
EPS (dextran) production by L. mesenteroides strains (Sánchez-Marroquín and Hope 1953; SánchezMarroquín et al. 1957; Chellapandian et al. 1998; García-Garibay and López-Munguía 1993; Ramírez et
al. 2004). However, after 6 hours of fermentation, Escalante et al. (2008) detected also a fructan EPS in
pulque. Screening for EPS producing Leuconostc species from aguamiel and fermented pulque, allowed
the isolation and identification of L. citreum as the unique EPS-producing bacteria (Conca 2008), questioning the essential role of L. mesenteroides during pulque fermentation.
43.6 Pulque: Nutritional and Traditional Medicine Aspects
Traditionally, pulque has been considered as a healthy beverage due to its nutrient content (described
above). Based on a daily consumption, pulque can be considered as an important source of energy, vitamins, and essential amino acids, outstanding content of the essential amino acids lysine and tryptophan,
which are deficient in the Mexican diet, which is based on maize. In some rural population in central
Mexico where potable water supply is absent, pulque is consumed as a water substitute due to its low alcohol content and high availability (Vargas 1999). The modern indigenous Otomíes living in the Mezquital
Valley include pulque as part of their basic diet; it represents the second source of energy (after tortillas)
and the third source of protein. Moreover, pulque is their main source of vitamin C and second of thiamine, riboflavin, calcium, and bioavailable iron (Vargas et al. 1998). Backstrand et al. (2002) in a study of
pulque consumption by several communities of the Estado de México highlands, established that a daily
intake of 500 mL supplied 215 kcal, 45% of vitamin C, 10% of niacin, 7% of thiamine, 6% riboflavin,
15% of iron, and microbial protein (with an adequate proportion of lysine and tryptophan).
Considering that pulque is also an alcoholic beverage, it has the same problems associated to excessive
consumption of nondistilled alcoholic beverages, but up to now, no toxicological problems other than
those related to excessive consumption of any alcoholic beverage has been reported for pulque. Since
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TABLE 43.1
Relevant Traditional Medical Uses and Beneficial Effects Associated with the Consumption of Pulque
Proposed Uses
Clinical Trial Evidence/Comments
Mixing with ground chili (Capsicum annuum
L.) and pumpkin seeds (Cucurbita spp.) as a
tonic
Boiled with chichicpactli (bark of Garrya
laurifolia Hartw. and G. ovata Benth.).
Beneficial for chest, stomach, and back pains
Control of: dyspepsia, stomach pain, diarrhea,
anemia, anorexia, asthenia, vertigo, severe
headache, and some neuralgias, urinal
infections, typhus, pityriasis, and syphilis.
Diuretic. It was consumed hot to favor
diaphoresis, relieve cough, and facilitate
expectoration. Sediments of pulque were
used in plasters to heal wounds
Increment of milk production during
pregnancy and lactation
None
Traditional pharmacopoeia
None
Traditional pharmacopoeia
None
Effects probably associated to
content of agave saponins (plants
sterols with hypocholesterolemic,
anti-inflammatory, and antibiotic
activity). Specific effect of pulque
remains unexplored
De la Cruz and Badiano 1964;
Peana et al. 1997; RamírezRancaño 2000
Possible galactogogue effect. Few
clinical trials related to the content
of ethanol in breast milk and effect
on the weight and size of infant at
moderate consumption
None
Argote-Espinoza et al. 1992;
Flores-Huerta et al. 1992;
Ortiz de Montellano 1993;
Backstrand et al. 2001, 2004
Avoid ferritin and hemoglobin
deficiencies due to a better
absorption and iron bioavailability
in the presence of ethanol and
vitamin C. In particular, the
consumption of pulque supplied the
Otomí women population with 7%
of their daily iron intake
None
Backstrand et al. 2001
Agaván (aguamiel syrup) against acute and
chronic failure of kidney, bladder, and
urethra; also for relief of cough and cold and
control of anemia
Control of anemia
Facilitate cellular respiration and absorption
of carbohydrates and proteins
References
Cook et al. 1995; RamírezRancaño 2000
Vargas et al. 1998
the middle of the 20th century, several authors have tried to correlate the high incidence of cirrhosis and
mortality in rural and indigenous communities in several central states of Mexico with pulque consumption. Nevertheless, it seems that there are other factors associated with the consumption of the beverage,
as the presence of endotoxins produced by enterobacteria, the deficient nutrition of the consumers, which
increase the rate of the disease or bad sanitary practices during its elaboration (Narro-Robles et al. 1992;
Vargas et al. 1998; Kershenovich 1999).
Since the pre-Hispanic period, pulque has been used as traditional medicine to control several diseases
(Table 43.1 summarizes principal traditional medical uses and beneficial effects associated to pulque
consumption). During the colonial period several medical applications of maguey and its by-products
were developed. Nevertheless, until very recently, the medical applications of pulque have been limited
to traditional pharmacopoeia. Actually, there is an extensive application of pulque in the treatment of
gastrointestinal disorders and intestinal infections that may be explained by pulque’s prebiotic and probiotic activities.
43.6.1 Pulque as Probiotic and Prebiotic Agent
Aguamiel and pulque can be considered as probiotic products due to the presence of LAB, as L. acidophilus and L. mesenteroides (Escalante et al. 2004, 2008; Campos 2010), Z. mobilis, which has an
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701
antagonist activity particularly against pathogenic bacteria and fungi species (Wuanick 1970; Gonçalves
de Lima 1978) and yeast (Steinkraus 1996) and as prebiotics because of its fructan and oligofructan
content (Ortiz-Basurto et al. 2008; Ramírez-Higuera 2009).
Several archeological evidences suggest that pulque has been used since pre-Hispanic times as an
enema. It has recently been proven that this practice may have had beneficial effects on humans’ health
due to the enrichment of the digestive tract with potential probiotic bacteria (Lemus 2006). In support
of the traditional applications of pulque in the treatment of several diseases, Campos (2010) reported the
isolation of several Leuconostoc species from pulque, with potential probiotic activities such as resistance to acid and bile salts and antimicrobial activity in vitro and in vivo against pathogenic bacteria.
Moreover, in recent experimental reports, Mancilla-Margalli and López (2006) and Ramírez-Higuera
(2009) demonstrated the prebiotic activity of aguamiel inulin since it increases the growth of LAB.
43.7 Pulque Industrialization and Major Technological Challenges
43.7.1 Pulque Fermentation Medium Composition
The carbohydrate structure of agave plants and aguamiel is rather complex, combining the fructose
polymer structure found in inulins and levans as described a few years ago by Mancilla-Margalli and
Lopez (2006). Actually, the term agavins has been proposed by these authors due to the clear structural
differences with inulin found in agave fructans. Most distilled agave fermented beverages rely on the
total hydrolysis of sugars, while pulque fermentation properties are based in the amount of sucrose and
monosaccharides found in aguamiel.
A modern production process requires the optimization and reduction in the agave production time.
Each agave pulquero is capable to generate up to 1000 L of aguamiel in a daily extraction basis during 90
to 150 days, with a sugar content of 7% to 14%, depending on the agave species. For instance, in the case
of Agave tequilana plants, the composition of water-soluble carbohydrates from plants of different ages
revealed that 2-year plants exhibited the highest level of free monosaccharide and low-molecular-weight
fructans (DP3–DP6) with potential application as prebiotics, while a maximum of fructan polymerization degree was achieved at 4 years with mean DP from 3 to 30 (Arrizon et al. 2009). According to these
authors, as the type and amount of sugars accumulated in A. tequilana plants varies with age, harvesting time should be optimized in terms of productivity and product quality, a conclusion that could certainly be extrapolated to other agave species. This is in agreement with statements made previously by
Sánchez-Marroquín (1970), who found the changes in aguamiel composition with collection time, agave
region, and agave varieties an additional inconvenient for the process industrialization.
Surprisingly, Ortiz-Basurto et al. (2008) detected only minor differences in aguamiel composition
among samples collected at different time intervals during the 3–6 months aguamiel harvest period of
an A. mapisaga plant. Actually, the analyzed aguamiel contained 11.5 wt% of dry matter, which was
composed mainly of sugars (75 wt%). Among these, 10 wt% were fructo-oligosaccharides. The variation of carbohydrate structure among the agave pulqueros, and its effect in the process and final product,
is a research topic that has not yet been addressed, including the acceptability of industrialized pulque
versions. On the other hand, industrialization has to be based in the planned and intensive production
(propagation) of selected agave species from the actual wide variety of genetic diversity cultivated in
México (García-Martín et al. 2007).
The introduction of hygienic aguamiel collecting and cajete-scraping techniques are also conditio sine
quibus non for modernization. Aseptic mechanical extraction systems as well as the proper collecting
and easy to handle reservoirs should be introduced. To avoid a preliminary fermentation, the agave juice
must be pasteurized by microfiltration or cooled, as fermentation will occur even in the agave basin if
not collected daily.
Up to now, no alternative to obtain aguamiel has been reported; however, it is always possible to obtain
agave stem extracts by pressing or by extraction in diffusors of the agave stems. This alternative was
reviewed previously (Steinkraus 1996), but no mention is made about the yields and sugar composition of
the resulting extracts. This possibility may require the hydrolysis of fructans present in the agave stem as in
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other agave fermentation procedures such as tequila, where the agave stems are either cooked (heat treated)
or ground and subjected to water extraction to obtain fermentable sugars, mainly fructose from agavins. The
4–6 months required for aguamiel extraction could then be reduced to a few hours, but the resulting media
although feasible for fermentation will no doubt differ from aguamiel and could not be referred as such.
43.7.2 Fermentation
The already described pioneer works of Alfredo Sánchez-Marroquín from 1953 to 1970 (Steinkraus
1996) are important not only in terms of the microbiological characterization of pulque, but also in
terms of efforts in the modernization of the production process. Sánchez-Marroquín was the first to
introduce a simplification of the enormous diversity of microorganisms isolated from the beverage to a
minimum starter formula composed of S. cerevisiae, Z. mobilis, Lactobacillus spp., and L. mesenteroides (Sánchez-Marroquín et al. 1957; Sánchez-Marroquín 1970). Although the three types of fermentations (alcoholic, lactic, and viscous) have been generally recognized as an essential element of pulque
production, the sensorial properties associated to the products of the three processes (alcohol, lactic acid,
and viscosity) can be obtained by alternative microorganisms. For instance, EPS associated to pulque
viscosity have always been recognized as dextrans (Sánchez-Marroquín and Hope 1953; Chellapandian
et al. 1998) but may also include levans and/or inulins (Morales Arrieta et al. 2006; Olvera et al. 2007;
Escalante et al. 2008). Isolated from pulque in 1924, Z. mobilis, recognized worldwide for its rapid alcohol fermentation, was included once among the essential pulque microbiota, but it is now considered as
a “casual microorganism for pulque fermentation” (Escalante et al. 2004).
Besides its ethanol production properties, Z. mobilis is also known to produce levansucrase, an enzyme
responsible for levan synthesis. Levan is a high-molecular-weight fructose polymer with β2–6 linkages
among the fructose molecules, which also induces high viscosity. It is interesting to point out that no
author has measured with precision the type and the amount of glycans (glucans and fructans) in pulque. However, and as already mentioned, it has been demonstrated that Leuconostoc spp. enzymatically
produce dextrans, levans, and/or inulins from the equivalent extracellular or cell-associated enzymes
(Chellapandian et al. 1998).
As early as 1967, Sánchez-Marroquín et al. reported that product quality as well as industrial operation
would improve if the viscous fermentation is avoided. Actually, to avoid the synthesis of EPS, it is possible
to invert all sucrose in aguamiel. Alternatively, glucose or fructose may also be added to support lactic
and alcoholic fermentations. Finally, the presence of strains such as L. mesenteroides may be avoided.
Nevertheless, efforts to preserve the prebiotic properties of fructo-oligosaccharides should be encouraged.
Although Sánchez-Marroquín et al. (1957) did not find differences in product quality following different inoculation strategies, parameters such as inoculation time and density of each of the selected strains
remains to be defined in terms of growth rate (LAB grow much faster than yeast), final lactic acid and
ethanol content, and eventually, EPS content. Kaffir (also known as Bantu), a traditional African beer, is
also a complex traditional fermentation beverage that has been successfully produced on a large scale.
In this case, the lactic acid fermentation of the saccharified sorghum starch takes place during one day,
before alcoholic fermentation, which requires an additional 2 to 5 days.
Modern pulque versions will first have to solve the quest that represents the microbial definition of pulque, or alternatively, promote the product resulting from a selected microbial inoculum. Lappe-Oliveras
et al. (2008) review the numerous reports dealing with pulque microbial studies in search for the optimal or essential microbial community; however, Escalante et al. (2008) demonstrated, using a polyphasic approach, to study bacterial diversity in aguamiel and pulque (including non–culture-dependent
approaches and PCR-based molecular techniques) that the microbial definition of pulque remains as a
difficult quest to solve.
43.7.3 Pulque Flavor
As far as flavor is concerned, Sánchez-Marroquín (1970) reports the measurement of an ester profile, but
no reference is given regarding the composition and profile of the pulque bouquet. The total amount of
esters (as ethylacetate) is reported as 20–30 mg/100 mL, aldehydes (as acetaldehyde) 2.5 mg/100 mL,
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703
and higher alcohols (fusel oils) 80–100 mg/L. It has been proposed that the common tradition of combining pulque with fruits or vegetables (curados) derives from the intention to correct off flavors or fermentation defects. Offensive flavors derived from bacterial contaminations, frequent in traditional pulque,
must absolutely be eliminated in a modern fermentation facility even if some traditional parts of the
process are conserved. Some undesirable flavor compounds often result from a combination of metabolic
pathways, particularly if a complex microbial diversity is involved. In a recent study, De León-Rodríguez
et al. (2008) reported the aroma compounds of agave alcoholic beverages as determined by gas chromatography and headspace solid-phase microextraction–gas chromatography–mass spectrometry as well
as a cluster analysis of the major components. There were 11 “major compounds” and around 17 minor
compounds that could be used as authenticity markers since they were beverage-specific and provide an
interesting fingerprint. It was observed that 3-methyl-thio-1-propanol and nonanoic acids are specific for
pulque and may be used as authenticity markers. In this study, pulque is grouped separately from others
agave beverages due to its low concentration of ethanol.
43.7.4 Pulque Conservation
The Mexican market in the United States and the ethnic food demands in the globalization era are the
major driving forces to develop a stable pulque product produced at a larger scale. One of the main challenges in this direction lies in the difficulties associated to conservation. As pulque is usually consumed
fresh after fermentation, mainly in rural areas, little efforts have been devoted to introduce stabilization
procedures. Therefore, there are many decisions regarding pulque processing after fermentation that
have to be taken into consideration, particularly if pulque production and commercialization continues
to rise. These decisions involve the introduction of pasteurization and/or filtration steps as well as the
addition of preservatives. If pasteurization had been introduced to pulque production at the same time
as in beer we would probably be reading a different story about the evolution of both beverages in the
20th century.
It is almost impossible to bottle fermented pulque without pasteurization, as sugars are usually not
exhausted; actually, a residual amount of agavin and fructo-oligosaccharides is desirable in order to
increase the prebiotic attributes of the product. On the other hand, filtration or sterilization would
increase pulque stability and shelf life but reduce its probiotic content, also recognized as one of pulque
nutritional attributes; efforts to preserve or supplement its vitamin (ascorbic acid) and nonheme iron
content should also have to be considered. Moreover, additives such as preservation agents, antioxidants, colorants, or texture agents will affect the image of pulque as a natural and, eventually, organic
product. In the particular case of the pasteurized bottled brand “Pulque, Cool Passion” (Poliqui SA de
CV), the product contains sodium benzoate as preservative, as well as color and sugars. The selection of
an adequate packaging requires consideration as biological and photochemical oxidation may also be a
problem in storage stability. The paradigm here is that to promote the industrialization of pulque, radical
changes in the public perception of pulque have to be made, particularly if new markets are searched.
An advantage is that the new pulque producers are probably offering a product that the large majority
of the public (particularly outside the country) never tasted before. Actually, it is highly possible that
new production processes will require modifications in the Mexican regulation system, as aguamiel as
well as pulque are defined specifically in terms of composition and production norms (NMX-V-022 and
NMX-V-037) approved in 1972.
According to the Del Razo group, which started canned pulque production in 1980 in the state of
Tlaxcala to fulfill the pulque demand in the United States, the market had already grown to more than
100,000 L/month by the start of the 21st century. The Del Razo pulque canning process includes aseptic
canning and a controlled process to preserve pulque’s sensorial and physicochemical properties. After
several unsuccessful efforts, the Del Razo group finally developed an effective process and now exports
to the United States the brand “Pulque Hacienda 1881” in various flavor presentations as well as natural
or plain pulque. According to Del Razo, the modification of their fermentation process has reduced the
traditional off-flavor associated with pulque and also its viscosity, which means that neither Z. mobilis
nor L. mesenteroides (nor at least the wild strains) are included in the process. Other trademarks include
Jicara, Malinche, Pulquemex, and Pulque Azteca, also competing for the North American market.
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Handbook of Plant-Based Fermented Food and Beverage Technology
After the paradigmatic work carried out by Sánchez-Marroquín, we are now witnessing a renewed
interest in this ethnic beverage from both basic and applied research. Pulque is now fully recognized as
rich in tradition, microbial diversity, and nutritional value. The recovery of “traditional pulque,” as well
as the birth of new alternatives of aguamiel processing are promoted by the demand of Mexicans living
abroad as well as the curiosity of consumers looking for new gastronomic and nutritional experiences.
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