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1
Microbial Valorization
of Tomato Waste for
the Extraction of
Carotenoids: Food
Applications
Praveen Kumar Dikkala
School of Food Technology, Jawaharlal Nehru Technological
University Kakinada (JNTUK), Kakinada, Andhra Pradesh,
India
Suman Biyyani
Department of Microbiology, Forest College and Research
Institute (FCRI), Mulugu, Hyderabad, Telangana, India
Gopinath Mummaleti
Department of Food Biotechnology, Indian Institute of Food
Processing Technology, Thanjavur, Tamil Nadu, India
Aparna Kuna
MFPI - Quality Control Laboratory, Prof. Jayashankar
Telangana State Agricultural University, Rajendranagar,
Hyderabad, Telangana, India
Pradeepa Roberts
Millet Processing and Incubation Centre, Prof. Jayashankar
Telangana State Agricultural University, Rajendra Nagar,
Hyderabad, Telangana, India
Kairam Narsaiah
AS & EC Division, ICAR-Central Institute of Post Harvest
Engineering and Technology, Ludhiana, Punjab, India
Chayanika Sarma
Department of Food Biotechnology, Indian Institute of Food
Processing Technology, Thanjavur, Tamil Nadu, India
DOI: 10.1201/9781003341307-1
1
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2
Microbial Bioprocessing of Agri-Food Wastes
Monika Sharma
Department of Botany, Shri Awadh Raz Singh Smarak
Degree College, Faizabad University, UP, India
Gauri Dutt Sharma
University of Science and Technology, Meghalaya, India
Zeba Usmani and Minaxi Sharma
Department of Applied Biology, University of Science and
Technology, Meghalaya, India
CONTENTS
1.1
1.2
1.3
Introduction.......................................................................................................2
Global Production and Valorization of Tomato Waste .................................. 4
Biochemistry of Different Bio-Active Components
from Tomato Pomace.......................................................................................5
1.4 Sustainable Valorization for Bioactive Components Extraction ....................6
1.4.1 Physico-Chemical Valorization Techniques........................................7
1.4.1.1 Conventional Solvent Extraction ..........................................7
1.4.1.2 Super Critical Fluid Extraction .............................................8
1.4.1.3 Pulsed Electric Field Extraction............................................ 8
1.4.1.4 Ohmic Heating Extraction.....................................................8
1.4.1.5 Ultra Sound and Microwave-Assisted Extractions...............9
1.4.1.6 Accelerated Solvent Extraction.............................................9
1.4.1.7 Hydrothermal Liquification .................................................10
1.4.2 Biotechnological Valorization............................................................10
1.4.2.1 By Microbial Fermentation .................................................10
1.4.3 Enzyme-Assisted Extraction .............................................................. 12
1.5 Applications of Carotenoids in Food Industry..............................................13
1.6 Future Scope and Conclusions....................................................................... 14
References................................................................................................................ 15
1.1 INTRODUCTION
From initial processing to final consumption, there are many phases in the food
chain. In each and every phase, food wastage is the major concern that causes
adverse impacts on nutritional security, environment, natural resources (Sharma
et al., 2021; Xue et al., 2017). The agricultural and food industrial wastes (AFIW)
have been reutilized by many industries, which could reduce industrial costs,
including capital costs (Pellegrini et al., 2018). Effective utilization of AFIW, which
are rich sources of different natural compounds, can be efficiently used as secondary
sources for developing value-added products (Sharma and Bhat, 2021). The
incorporation of valuable components obtained from food waste in food formulations for the improvement of nutritional quality is an emerging area of research
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Microbial Valorization of Tomato Waste
3
(Herrera et al., 2010). One such AFW source is tomato (Solanum lycopersium)
waste that includes pomace, peel, and seeds, which is the second-most widely
produced vegetable crop globally and consumed either directly (fresh tomato) or
used for processing different tomato products such as puree, juice, concentrate,
sauce, soup, ketchup, whole dried tomatoes, and tomato powder. Tomatoes and
tomato-based products provide greater than 85% of the total lycopene intake to the
human body (Amiri-Rigi et al., 2016). The global production of tomatoes is about
180 million tonnes approximately of which about 39 million tonnes are going to
industry to process (https://www.tomatonews.com/en/background_47.html). From
tomatoes, the global revenue amount is about $190.4 billion in the year 2018, which
was raised by 6.5% in 2019 (Amiri-Rigi et al., 2016). These numbers reflect the
total revenues of both the exporters and importers (excluding the retail market costs
and logistics costs) (Global Tomato Industry Report, 2020).
Approximately 40 million tonnes of tomatoes are processed annually generating
tomato pomace (byproduct), which is a mix of vascular tissues, tomato peels, and
seeds and a small fraction of the pulp (Szabo et al., 2018). Around 5–30% of the
main product is produced as a byproduct. The amount of pomace produced varies
depending on the raw material and processing conditions, and it has been reported
that an amount of 600 thousand to 2 million tonnes of tomato pomace is produced
(Yasmin et al., 2020). Tomato pomace consists of nearly 33% of seed, 27% of skin,
40% of pulp, in the dried form 56% of skin and pulp and 44% of seed (Poojary and
Passamonti, 2015; Ruiz-Celma et al., 2012). Tomato byproducts have significantly
high amounts of dietary fiber and bioactive phytochemicals like anthocyanins,
sterols, terpenes, polyphenols, and carotenoids (Kalogeropoulos et al., 2012).
Chanforan et al. (2012) reported that the overall nutritional quality of tomato
products did not decrease, except vitamin C, with industrial processing and also
during storage. Tomato pomace particularly skin contains the highest amount of
lycopene (Papaioannou and Karabelas, 2012; Strati and Oreopoulou, 2014). Apart
from lycopenes and carotenoids, tomatoes are abundant sources of tocopherols,
terpenes, sterols, and polyphenols (Kalogeropoulos et al., 2012). The extracts
obtained from tomatoes (pomace) are the richest sources of phenolic antioxidants,
particularly heat- and oxidation-resistant compounds (Ćetkovicć et al., 2012). Major
studies on tomato pomace are on carotenoid characterization (lycopenes and
β-carotenes). In the extraction of carotenoids, seasonal production plays an important
role, with the highest proportions of lycopene and β-carotene during the summer
season (Riggi and Avola, 2008). Considering the high potential of tomato wastes,
recovery of nutrients and antioxidant bioactive compounds can also contribute to
improved nutritional security by reducing degenerative diseases caused by oxidative
damage and cancer. Moreover, in-vivo studies are being performed to analyze the bioavailability and real benefits of these tomato extracts (Stajcic et al., 2015). The wastes
from tomato industries are creating environmental issues although they are valuable
sources of different bioactive components (Szabo et al., 2018). Prior to consideration
as a nutraceutical source, it is very important to explore its environmental and economic sources (profit analysis). Different valorization techniques such as ultrasonic,
green-solvent, microwave, or supercritical fluid extraction can be adopted for the
extraction of different bioactive components from tomato wastes. The aim of this
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Microbial Bioprocessing of Agri-Food Wastes
FIGURE 1.1 Schematic representation of valorization of tomato waste and byproducts for
carotenoids and their potential applications.
chapter is to present an overview of the valorization techniques in detail. Figure 1.1 is
showing valorization technologies for tomato waste and their valuable applications.
1.2 GLOBAL PRODUCTION AND VALORIZATION OF TOMATO
WASTE
Tomatoes are one of the most significant vegetables used worldwide because of
their taste, nutritional value, different forms, and colors. Currently, the global
production of tomatoes were around 164 million tons approximately showed by
Food and Agriculture Organization Statistics (FAOSTAT) (Amiri-Rigi et al., 2016).
From tomatoes, the global revenue amount is about $190.4 billion in the year 2018,
which was raised by 6.5% in 2019. These numbers reflect the total revenues of both
the exporters and importers (excluding the retail market costs and logistics costs)
(Global Tomato Industry Report, 2020). Tomatoes are the most cultivated vegetable
with 17.9 million tons of production in the European Union in 2016. Tomatoes and
tomato-based products provide greater than 85% of lycopene intake in the human
body (Amiri-Rigi et al., 2016). EUROSTAT (The Statistical Office of the European
Union) stated that greater than 10 million tons of tomatoes were processed to produce
different varieties of products (ketchup, pastes, puree, sauces, canned tomatoes). After
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Microbial Valorization of Tomato Waste
5
industrial processing, large amounts of waste are generated includes, vascular tissues,
tomato peels, and seeds etc. (Szabo et al., 2018). Tomato pomace, consists of 33%
of seed, 27% of skin, 40% of pulp, in the dried form 56% of skin and pulp, and 44% of
seed (Poojary and Passamonti, 2015).
1.3 BIOCHEMISTRY OF DIFFERENT BIO-ACTIVE COMPONENTS
FROM TOMATO POMACE
Comparative studies were done on processed and raw tomatoes for different phytochemical substances (Kalogeropoulos et al., 2012). Different phytochemicals
(tocopherols, terpenes, polyphenols) and bioactive components (carotenoids) were
present in huge amounts, even after processing those which are able to withstand
processing conditions industrially. More than 600 carotenoids are naturally present
in fruits, vegetables, fungi, bacteria, and algae and are divided into lycopene,
xanthophylls, and carotenes based on the functional group. Carotenoids are also
hydrophobic with polyene chain that changes the polarity of carotenoids, influences
biological membranes and molecules. The carotenoids in plant tissues dissolve in
oily solvents and are esterified with sugars, proteins and fatty acids. Carotenoids are
divided into hydrocarbon carotenoids and xanthophylls based on the presence of
oxygen in their structure (Story et al. 2010). The non-oxygenated carotenoids are
carotenes including α-, β-, & γ-carotenes, lycopene, phytofluene, and phytoene. The
oxygenated carotenoids are xanthophylls that contain oxygen in hydroxyl, keto,
carboxy, methoxy, and epoxy groups (Oliver and Palou 2000). The carotenoids are
yellow to red in color that belongs to the tetraterpenes group. The most important
bioactive components present in industrial tomato byproducts are carotenoids,
lycopene along with some amount of zeaxanthin, phytofluenes, neurosporenes,
luteins, α-carotenes, β-carotenes, gamma-carotenes, and ξ-carotenes (Szabo et al.,
2018). Carotenes are composed of eight isoprenoid units in a carbon chain backbone
having alternative double bonds with cyclic/acyclic functional groups (GómezGarcía and Ochoa-Alejo, 2013). The conjugated double bonds of the compounds
are attributed to their antioxidant activity by which these compounds can scavenge
the free radicals to make them stabilize wherever required.
The conjugated double bonds of the carotenoids are responsible for the characteristic color of materials and regulate various biological functions such as photosynthesis, energy transfer, protection from light, etc. As the carotenoids are precursors
of vitamin A, α-carotene and γ-carotene are having the ability to synthesize one
molecule of vitamin A, whereas one β-carotene molecule can produce two molecules
of vitamin A (Sajilata et al., 2008). Lycopene is an aliphatic carotenoid found in
tomatoes, grapes, and watermelons (Engelmann et al., 2011). Lycopene is generally
found in trans form chemically, which is a quite stable form, so it is important to avoid
cis-trans isomerization reactions while incorporation of lycopene in food formulations. The peel, pomace, and seeds generated from the processing industry can be a
feasible wellspring of lycopene as the skin only contains five times more lycopene per
unit of mass as compared to its pulp. Lutein carotenoids contain an alcohol group
in their structure with hydroaromatic α structure (Mikami and Hosokawa 2013). It is
a dihydroxy carotene with ionone rings carrying hydroxyl groups. Astaxanthin is a
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Microbial Bioprocessing of Agri-Food Wastes
FIGURE 1.2 Biochemical structures of various carotenoids.
metabolite of zeaxanthin a keto carotenoid with hydroxyl and ketone groups. The
lycopene and β-carotene contents in the dried tomato wastes were about 510.6 mg/kg
and 95.6 mg/kg, respectively. The total phenolic content and the flavonoid content
were recorded to be about 1229.5 mg GAE/kg and 415.3 mg QE/kg, respectively
(Nour et al., 2018). The structures of various carotenoids were shown in Figure 1.2.
1.4 SUSTAINABLE VALORIZATION FOR BIOACTIVE
COMPONENTS EXTRACTION
Tomato peel, pomace, and seeds as tomato waste can be valorized in different ways for
the recovery of different value-added ingredients due to differences in chemical
composition. The functional ingredients that are developed from tomato wastes are
mainly used for the development of functional and nutraceuticals foods. The valorization of the waste is a holistic approach that envelope several sequential technologies
such as pretreatment, extraction, purification and isolation, encapsulation, and
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Microbial Valorization of Tomato Waste
7
TABLE 1.1
The Comprehensive Utilization of Tomato Pomace for the Production of
Lycopene
Residue of
tomato
Extraction Process
Yield
Reference
Pomace
Solvent extraction
119.8 (wet)
Al-Wandawi et al. (1985)
Pomace
Solvent extraction
820 (dry)
Machmudah et al. (2012)
Pomace
Pomace
Supercritical fluid extraction
Supercritical fluid extraction
459 (dry)
314 (dry)
Machmudah et al. (2012)
Vagi et al. (2007)
Pomace
Solvent extraction
734 (dry)
Knoblich et al. (2005)
Pomace
Pomace
Solvent extraction
Supercritical fluid extraction
24.5 (dry)
14.86 (dry)
Rozzi et al. (2002)
Rozzi et al. (2002)
Pomace
Supercritical fluid extraction
31.25 (wet)
Yi et al. (2009)
Pomace
Pomace
Solvent extraction
Solvent extraction
19.8 (dry)
739 (wet)
Kaur et al. (2008)
Lavelli and Torresani, (2011)
Pomace
Solvent extraction
6.07 mg/100 g
Perretti et al. (2013)
Pomace
Pomace
Sunflower oil as green solvent
Solvent extraction
2.59 mg/100 g
5.22
Perretti et al. (2013)
Yilmaz et al. (2016)
Pomace
Ultrasound-assisted extraction
7.01
Yilmaz et al. (2016)
Pomace
Pomace
Supercritical fluid extraction
Supercritical fluid extraction
72.90
45.92
Kehili et al. (2017)
Machmudah et al. (2012)
Pomace
Supercritical fluid extraction
31.72
Vagi et al. (2007)
Pomace
Pomace
Supercritical fluid extraction
Enzyme-assisted extraction
28.26
2.30
Huang et al. (2008)
Azabou et al. (2016)
Pomace
Ethyl lactate-enzyme-assisted extraction
8.94
Strati and Oreopoulou (2014)
Pomace
Ethyl lactate- green solvent extraction &
high hydrostatic pressure extraction
8.36
Strati and Oreopoulou (2014)
incorporation in functional food formulations. The extraction potential of different
organic solvents was examined to optimize the extraction parameters for maximum
yield (solvent form, extraction time, temperature, and extraction steps) (Strati and
Oreopoulou, 2011). Lycopene content of peel is about five times more than as compared with its pulp per unit of mass. Tomato pomace and tomato skin are the richest
sources of lycopene and other carotenoids. Different types of valorization techniques
like physico-chemical, enzymatic, and biotechnological techniques can be adopted to
process the tomato pomace, as shown in Table 1.1 (Baiano and Del Nobile, 2016).
1.4.1 PHYSICO-CHEMICAL VALORIZATION TECHNIQUES
1.4.1.1 Conventional Solvent Extraction
Different chemical waste valorization techniques were used to extract several different bioactive ingredients from tomato pomace. Amongst them, solvent extraction
is one of the most common and traditional one. Various types of organic solvents
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Microbial Bioprocessing of Agri-Food Wastes
such as hexane, petroleum ether, isopropyl alcohol, methanol, cyclohexane, di-ethyl
ether, etc. are used for the extraction of carotenoids as they are non-polar compounds. Solvent type is the key parameter in the process of solvent extraction. With
the optimization of different extraction parameters such as solvent type, time and
temperature of extraction, extraction steps, extraction time, size of the particle,
moisture content, etc. the extraction potential of different organic solvents was
enhanced (Strati and Oreopoulou, 2011). Different researchers gave the conclusion
that ethyl lactate gives the highest carotenoid content (Ishida and Chapman, 2009;
Kaur et al., 2008). The process of solvent extraction is the commonest method in
carotenoid recovery because of its high hydrophobicity and less water solubility. For
the extraction of carotenoids and other bioactive ingredients, Soxhlet extraction and
agitation are the major techniques. The solvents that are used for extraction should be
non-toxic for human health. But, the organic solvents used in the solvent extraction
process have adverse effects on human health that cannot be removed entirely from
the extracts. Therefore, research for alternative solvents with very less negative
impacts should be conducted (Ho et al., 2015). This problem was solved by the use of
edible oils as solvents and deep eutectic solvents, generally called green solvents, and
also by utilizing innovative extraction technologies.
1.4.1.2 Super Critical Fluid Extraction
It usually depends upon the properties of the fluids. With the increase in pressure and
temperature greater than the critical point, the solvating power of the gas is increased
(Cadoni et al., 1999). The most commonly used compound in super-critical fluid
extraction is carbon dioxide because of its low critical temperature and pressure.
Carbon dioxide is the most favored alternative to organic solvent due to its properties
(inexpensive, non-toxic, non-explosive). The liphophilic substances can be easily
solubilized with the use of carbon dioxide. Several studies showed that super- critical
fluid extraction is best for the recovery of carotenoids from the tomato pomace and
wastes (Baysal et al., 2000). With the increase in pressure and temperature of carbon
dioxide, the extracted amount of lycopene is increased. With the increase in density of
supercritical carbon dioxide, the number of carotenoid solubilization is increased.
1.4.1.3 Pulsed Electric Field Extraction
A combination of solvents (hexane: acetone: ethanol) and pulsed electric field (PEF)
technology, the extraction of carotenoids increase, along with decreased usage of green
solvents. The permeabilization of tomato pomace at different electrical field strengths
was about 90 µs, according to the cell disintegration index. PEF permeabilization did
not increase the output of carotenoids from tomato pulp, whereas it increased by 39% in
the peel when compared with the control at PFE treatment (5 kV/cm) Addition of
acetone mixture with solvent did not affect the carotenoids extraction positively after
the treatment with PEF, but this process reduced the hexane utilization from 45% to 30%
without any negative impact on the carotenoid’s extraction (Luengo et al., 2014).
1.4.1.4 Ohmic Heating Extraction
In the food industry, solvent extraction with different organic solvents and polar or
non-polar combinations have been assessed for carotenoid extraction (Strati and
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Microbial Valorization of Tomato Waste
9
Oreopoulou, 2011). It is very difficult to apply ohmic technology to non-conductive
and non-homogenous food systems. In addition, many systems/foods are rich in
proteins that can come up with the formation of deposits on the surface of OH
electrodes, which can result in an electrical arc if not properly cleaned (Kumar,
2018). The above-mentioned drawbacks can be easily managed by using other
greener solvents for the extraction of carotenoids from tomato processing waste. It
seemed promising to extract lycopene from tomatoes by means of vegetable oils,
environmentally friendly solvents, rather than more harmful organic solvents
(hexane, chloroform, petroleum ether) because of its fat-soluble and environmental
concern (Lenucci et al., 2015).
1.4.1.5 Ultra Sound and Microwave-Assisted Extractions
These are promising greener strategies for improved extraction of lycopene when
compared with conventional methods for valorization of tomato pomace. The
ultrasound waves and microwaves modify the structure of the cell wall due to
electromagnetic waves and are adapted for the extraction of lycopene from tomato
waste (Kusuma and Mahfud, 2016). The combination of technologies are better
than conventional techniques, in terms of less environmental pollution, reduction in
solvent usage, and more extraction in a short time. However, certain disadvantages
like the usage of additional filtration for the removal of solid residues are also
reported. The microwave might be influenced by the volatile compounds (Ho et al.,
2015; Baiano et al., 2014). Ultrasound-assisted extraction of lycopene from tomato
wastes is eco-friendly using solvents like ethyl lactate, ethyl acetate, with enhanced
extraction. Silva et al. (2019) reported improved carotenoids yield up to 125.3 µg/g
using ultrasound-assisted extraction. The ultrasound treatment increased the yield of
extractable lycopene and it is clearly showed as an alternative source for extractable
lycopene.
1.4.1.6 Accelerated Solvent Extraction
The extraction technology used for bioactive components with the application of
temperature and pressure ranging from 50°C to 200°C and 9–15 Mpa, respectively,
is called as accelerated solvent extraction or pressurized liquid extraction. In this
extraction technique, the extractant solvent is in liquid state synergizes with the
high temperature which influences the enhanced extraction of bioactive components
like lycopene. The solubility of the bioactive components was enhanced due to the
solvents that are forced into the matrix. With the process of accelerated solvent
extraction, Naviglio et al. (2008), extracted the lycopene at 0.7–0.9 MPa pressure
using tap water as an extracting medium from tomato pomace obtained from a
processing industry. The molecular aggregates of lycopene and water were extracted using pressure and depressure cycles. Another technique i.e., high hydrostatic pressure extraction at 100–800 MPa pressure with low temperature (room
temperature) is also gaining huge attention in extraction studies of carotenoids from
food wastes, especially from tomato processing wastes. Some studies showed that
higher hydrostatic pressure extraction yields more amount of bioactive components
when compared to conventional extraction technologies (Strati et al., 2014). High
hydrostatic pressure extraction is more advantageous over conventional extraction
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Microbial Bioprocessing of Agri-Food Wastes
methods as it consumes less time, uses less solvent, gives more yield, and has more
convenient technology that can be operated at room temperature (Zhang, 2015).
1.4.1.7 Hydrothermal Liquification
Hydrothermal liquification plays an important role in biomass conversion technologies. This technique is most commonly used to renovate the inedible food waste
biomass into oily compounds that have further applications to produce valuable
chemicals such as fatty acids, phenols, and polyols, which are valorized into different
biofuels like biodiesel (Zhang, 2015).
1.4.2 BIOTECHNOLOGICAL VALORIZATION
Biotechnological processes play an important role in tomato waste valorization. Due
to the huge moisture content present in the tomato pomace, it cannot be stored for
more than 6 to 7 days due to its putrescible nature. With different results from
the fermentation tests and adapted parameters, it was clearly concluded that tomato
byproducts are considered a low-cost potential alternative source for bioethanol
production with optimization process. Usually, residues from tomato processing
industries increase the cost of disposal (Bacenetti et al., 2015). Not only this,
uncontrolled anaerobic fermentation releases more methane that impacts the tropospheric zone in the atmosphere (Bacenetti et al., 2015). Hence, valorization of tomato
pomace is a safe alternative to reduce their negative impacts on the environment.
Cascade fractionation (innovative extraction process) plays an important role in the
extraction of different valuable industrial byproducts from tomatoes. Various extraction products such as carotenoids, oleoresins, tomato seed oils, proteins, serve as a
good source of lignocellulosic material for bioethanol production (Kehili et al., 2017).
Different lactic acid bacteria such as Lactobacillus species, yeast strains such as
Saccharomyces boulardii, S. cerevisiae, and S. boulardii improved the nutritional
value of the different tomato byproducts and described in Table 1.2. Fermentation of
tomato pomace with lactic acid bacteria and yeasts improves the nutritional value in
terms of minerals such as calcium, magnesium, potassium, iron, enhanced energy,
protein content, fat, and titrable acidity. However, fermentation has a negative impact
on pH and TSS (total soluble solid). Tomato pomace fermentation with lactic acid
improves nutritional quality with a reduction in fiber content, which is a good source
of animal feed supplement (Roja et al., 2017). With proteolytic bacteria (Bacillus
subtilis), the antioxidant and antimicrobial hydrolysates were produced with the
fermentation of protein fractions of tomato seeds (Moayedi et al., 2016).
1.4.2.1 By Microbial Fermentation
The fermentation processes, not only improved the extraction yield of the bioactive
components but also enhance the stability and bioavailability of carotenoids (betacarotene, lycopene, and astaxanthin) and improve the cleavage of their derivatives
(Mapelli-Brahm et al., 2020). The tomato wastes were fermented at different
temperatures from 15 to 20°C with Saccharomyces bayanus, BV 818 at a concentration of 0.03 to 100g (Owusu et al., 2014). They reported that the pH and
temperature during fermentation had a great influence on the extraction efficiency
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Microbial strains
Fusarium solani pisi
Pediococcus
acidilactici
Lactobacillus sakei
Fusarium solani pisi
Rhodotorula glutinis
Rhodotorula glutinis
Pediococcus
pentosaceus
Saccharomyces
bayanus
Tomato waste
Tomato peels
Tomato pulp
Tomato pulp
Tomato seeds
Tomato waste
Tomato extract
Tomato pulp
Tomato pomace
Temperature: 15, 20 °C; pH-4.11
Temperature: 28°C; 150 rpm
Temperature: 30–35°C
Temperature: 30°C; 120 rpm; 72 h
Temperature: 30–35°C
Temperature: 37, 45 and 50°C;
pH-4.5, 5, 8, and 9
Temperature: 30–35°C
Temperature: 37, 45 and 50°C;
pH-4.5, 5, 8 and 9
Processing conditions
Hexane:Acetone:Ethanol in
10:5:5 ratios
Acetone
Hexane:Acetone in 1:1 ratio
Petroleum ether
Hexane:Acetone in 1:1 ratio
Ethanol
Hexane:Acetone in 1:1 ratio
Ethanol
Solvent extraction
3.06
13.43
5.68
3.64
5.68
2.7
5.68
2.7
Yield
(mg/100 g)
Owusu et al. (2015)
Wang et al. (2007)
Bartkiene et al. (2013)
Chandi et al. (2010)
Bartkiene et al. (2013)
Azabou et al. (2016)
Bartkiene et al. (2013)
Azabou et al. (2016)
Reference
TABLE 1.2
The Different Extraction Technologies using Microbial Fermentation of Tomato Waste for the Production of Carotenoids
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Microbial Bioprocessing of Agri-Food Wastes
and yield of lycopene and beta-carotenes. The total antioxidant levels and reducing
powers were highest at 3.2 pH and 20°C temperature (Owusu et al., 2015). A
research study by Jamal et al. (2017) showed that lycopene extraction was improved
using solid-state fermentation with Aspergillus niger, whereby the cellulases produced after fermentation degraded the cell-wall constituents that facilitated the
release of intracellular contents. There was some influence of independent factors
such as moisture content (60–80%), inoculum size (5–15%), and incubation time
(2, 3, and 4 days) on the recovery of the carotenoids. The optimized conditions for
maximum extraction of lycopene were moisture content of about 80%, inoculum
size – 15% after 4 days of incubation.
Another study by Kim et al. (2010) reported improved lycopene recovery from
tomato wines (4.2 mg/100 g), due to the effect of alcoholic fermentation using different yeast strains (Saccharomyces cerevisiae and S. bayanus lalvin EC-1118 sp.)
and sugars. Lycopene concentration was reported at about 45 mg/100 g of tomato
wines with optimal fermentation conditions and bio-accessibility of lycopene was
improved (Lu et al., 2020). In tomato byproducts, seeds are the richest sources of
proteins. Fermentation of tomato seeds with Lactobacillus plantarum degraded the
tomato seeds into bioactive peptides having the capability to improve the antioxidant
activities of protein isolates (Shehzadi et al., 2018).
1.4.3 ENZYME-ASSISTED EXTRACTION
The low-cost commercial food-grade enzymes can be used in the extraction of
carotenoids with the possibility to improve the efficiency and yield of the targeted
compounds from the lab scale to the industrial level. Optimization of different
pretreatments (heat-treated dilute acid and ultrasound-assisted dilute acid) and enzymatic methods could be adapted to extract the highest amount of fermentable
sugars with tomato pomace. Enzymatic pretreatments for extraction of bioactive
components from tomato pomace contain many advantages; it reduces the time and
solvent consumption and also enhances the quality and quantity. However, there are
some limitations to this technique, such as the cost of enzymes, enzyme preparations for complete hydrolysis, and industrial feasibility for different conditions (Puri
et al., 2012; Zuorro et al., 2011). From these studies, it was observed that lycopene
recovery was enhanced greatly by the use of mixed enzymatic preparations with
different low-cost commercial food-grade enzymatic preparations with optimum
cellulolytic and pectinolytic activities and the possibility of implementation at the
industrial level.
The cellulase enzymes optimization showed an improvement in the fermentable
sugars in tomato pomace. Conditions optimized for the extraction of sugars were good
with 1.5% cellulase enzyme used at 6 h of incubation. Along with this, the application
of pretreatments (heat-treated dilute acid) and enzymatic hydrolysis process on
tomato pomace improved the ester and alcoholic compounds, which have applications
in different food, cosmetic, chemical, and pharmaceutical companies. Another study
by Zuorro et al. (2011), reported the improvement in lycopene recovery using a
mixture of two enzymatic preparations, used for the extraction from tomato
peel waste. The lycopene content was improved from 8- to 18-fold with 50:50% of
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pectinolytic and cellulolytic enzymes mixture. In this process, the extraction optimized conditions were temperature at 30°C, extraction time 3.18 h and enzyme
mixture load was about 0.16 kg/kg of peel waste. The study showed that the use of cell
wall degrading enzymes was a promising approach to improve the recovery of
lycopene from tomato peel waste.
1.5 APPLICATIONS OF CAROTENOIDS IN FOOD INDUSTRY
Visual aesthetics and appearance are the most important factors for the successful
acceptability and marketability of the product, as they give the first impression to
consumers about the products. The use of food additives particularly colorants have
strict regulations in many countries. Most of the consumer complaints regarding the
food industry are mainly about the use of synthetic food colors that affect the health
of consumers. The demand for natural food additives such as colorants is rising due
to an increase in awareness of consumers towards natural products rather than
chemical additives (Santos et al., 2011). The biosynthesis of carotenoids from
microorganisms was gaining importance and competing with the chemical synthesis
procedures. The global market for the pigments produced from microorganisms was
increasing when compared to the chemically synthesized pigments (Strati and
Oreopoulou, 2014). In the food processing industry, the carotenoids are extracted to
use them as colorants in juices, beverages, confectioneries, margarine, sausages,
and cheeses. The foods fortified with carotenoids are mainly due to their coloring
property and health benefits such as antioxidant activity. The red color of tomatoes
is mainly due to the presence of lycopene, a commercially important potential
natural dark red color pigment. The carotenoids are present mostly in darkpigmented fruits and vegetables. The β-carotene is the best-known food carotenoids
and found along with α-carotene in some foods. Lutein and zeaxanthin are isomers,
that help in the prevention of macular degeneration. Unique carotenoids like bixin
are found in annatto and crocin is found in saffron (Rodriguez, 1999). The lycopene
of tomatoes is stable to extreme temperatures, pH and these are effective at low
concentrations. Oleoresin and tomato pulp powder were used for color stabilization
and for preventing oxidative degradation in beef patties by Sánchez-Escalante et al.
(2003). They reported that the oleoresin was found to be highly effective against
lipid oxidation. The tomato powder was incorporated in pork patties by Kim et al.
(2013) and in beef burgers by Luisa García et al. (2009), and the results revealed
lower thiobarbituric acid values than other control samples. The patties incorporated
with tomato powder showed high redness and low discoloration rate when compared to control. The sensory scores were also higher for patties incorporated with
tomato powder than other samples. The incorporation of carotenoids in food
products such as macaroni not only improves the nutritional and organoleptic
acceptability but also reduces undesirable reactions during processing and consumer
intake (Ajila et al., 2010). The firmness of the product will be maintained even after
processing and cooking losses.
Astaxanthin extracted from muscle proteins of shrimp and lycopene from tomato
were extracted and incorporated in edible films to develop antioxidant edible films.
The films on storing for 1 month recorded 32% of lycopene and 17% of astaxanthin
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Microbial Bioprocessing of Agri-Food Wastes
degradation, but the antioxidant activity was found to be stable (Mohd Hatta and
Othman, 2020). The effect of carotenoids extracted from industrial tomato wastes on
the enrichment of edible oils was studied and reported that refined oil had better
thermal stability followed by extra olive oil and sunflower oil (Benakmoum et al.,
2008). Encapsulation of carotenoids is another important technique that helps to
improve the usage of carotenoids. Encapsulation of astaxanthin with cashew gum for
forming coacervate with gelatin and stability was studied by Martins and Ferreira
(2017). The results showed that the coloring capacity and stability increased with
encapsulated carotenoid than with the non-encapsulated carotenoid. Carotenoids have
several biological functions and their nutritional importance and health benefits were
well studied. The carotenoids protect the cells and tissues from oxidative damage,
enhance the immune system, protect from sunburn, and also protect from some
cancers. Carotenoids also prevent oxidation of low-density lipoprotein thereby
reducing coronary heart diseases and atherosclerosis (Alem Zeb et al., 2004).
1.6 FUTURE SCOPE AND CONCLUSIONS
Carotenoids are highly valuable components that are wasted in the form of AFW if
left unextracted. Several studies on the beneficial effects of carotenoids created
interest in researchers to explore carotenoids. On the other hand, interest in natural
food additives particularly colorants is increasing every day due to the health
benefits of natural colorants in comparison with synthetic additives. The market for
carotenoids is increasing globally for their application in food, pharmaceutical,
cosmetic, beverage, nutraceutical and animal feed industry.
The tomato processing industry produces huge amounts of byproducts with
potential bioactive components, which are healthy, and therefore are in great demand
by food industry. Lycopene is the major carotenoid present in tomato which has a
huge demand in food processing as a colorant and antioxidant. Extraction of carotenoids is usually done using organic solvents due to the lipophilic character of the
carotenoids, but these organic solvents have a high impact on the environment, apart
from their residual presence in the final end product, which is toxic for human consumption. This problem was solved by the use of edible oils as solvents and supercritical fluid extraction techniques. However, these techniques were found to be costly
and extraction parameters should be selected carefully to achieve higher yields of
carotenoids with high antioxidant activity. To improve the extraction rate, yield,
reduce the time, reduce the cost, and protect the thermolabile components, few
supporting methods such as high hydrostatic pressures, microwave, ultrasound, and
radiofrequency techniques are gaining momentum alone or in combinations. These
green extraction techniques need to be further developed to extract carotenoids of
high quality with good yield, stability, and activity along with low cost and low hazard
to the environment. Limited studies reported the incorporation of carotenoids to
develop active packaging materials and edible films to extend the shelf life of
products, which can be further explored. Rapid increase in the field of genetic engineering and biotechnology helps in producing the carotenoids in higher amounts
naturally by using microorganisms. Genetic engineering techniques can be employed
and explored for the green synthesis of carotenoids in higher amounts using microbes.
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The cultivation of microorganisms is easier and higher yields can be achieved in less
time. The extraction of carotenoids from natural algae and fungi needs to be explored
as a potential source. The fermentation process can make many complex modifications in the substrates and also produces several bioactive compounds in the process
of fermentation. The fermentative production of carotenoids using agro and food
industry waste as low-cost substrates need to be exploited to enhance the production
of carotenoids without affecting the environment.
Encapsulation of carotenoids, using several synthetic gums is also gaining
importance due to the improved stability and activity of the encapsulated material.
However, the use of natural compounds such as protein and carbohydrate formulations as wall materials for encapsulation of carotenoids, along with absorption,
bioavailability, and bioaccessibility of encapsulated carotenoids needs further exploration. The incorporation of carotenoids in food systems and the stability of
carotenoids during the processing and storage of food products need to be studied.
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