Application of Bacteriocins in
food products
1
Present by
Nirarat Thongchai 55010219071
What is Bacteriocin ?
 Bacteriocins are ribosomally -synthesized peptides or
proteins with antimicrobial activity.
 Produced by different groups of bacteria.
- Gram-positive bacteria.
- Gram-negative bacteria.
- Archaea.
 The main differences between bacteriocins and
antibiotics are summarized in Table
Cleveland et al.(2001)
2
Bacteriocins vs. antibiotics
Characteristic
Application
Bacteriocins
Food
Antibiotics
Clinical
Synthesis
Ribosomal
Secondary metabolite
Activity
Narrow spectrum
Varying spectrum
Host cell immunity
Yes
No
Mechanism of target cell
Usually adaptation affecting
cell
Usually a genetically
transferable
resistance or tolerance
membrane composition
determinant affecting different
sites
depending the mode of action
Interaction requirements
Sometimes docking molecules
Specific target
Mode of action
Mostly pore formation, but in a
few cases possibly cell wall
biosynthesis
Cell membrane or intracellular
targets
Toxicity/side effects
None known
Yes
Cleveland et al.(2001)
3
Classification of bacteriocin
Bacteriocins of Gram-positive bacteria
Class I
Lantibiotic
Class II
Unmodified peptide
Class III
Large Proteins
Type A
Linear
Type A
pediocin-like
Type A
Bacteriolytic
Type B
Globular
Type B
Miscellaneous
Type B
Non- lytic
Type C
Multicomponent
Type C
Multicomponent
Class IV
Cyclic peptides
Cotter et al. (2006),
Klaenhammer (1993).
4
Bacteriocins of Gram-negative bacteria
Type
Size
Examples
Pore Formers
Nucleases
(20-80 kDa)
Colicins
Colicin-like
(20-80 kDa)
S -pyocins
Klebicins
Phage-tail like
(>80 kDa)
R and F pyocins
Posttranslationally
modified
Unmodified
(<10 kDa)
Microcins
References
(Cascales et al. 2007)
(Michel-Briand and Baysse
2002)
(Gillor et al. 2004)
(Reeves 1965)
Bakkal et al, 2012
5
Bacteriocins of Archaea
Halobacterium
Sulfolobus
(extreme halophiles)
(exthreme thermophilies)
Halocins
• Microhalocins (< 10 kDa)
• Protein halocins (>10 kDa)
Sulfolobicin
Bakkal et al, 2012
6
7
Modes of action of Bacteriocins from
Gram-positive bacteria
Cotter et al. (2006)
8
Modes of action of Bacteriocins from
Gram-negative bacteria
Cotter et al. (2006)
9
10
Bacteriocins-producing Gram-positive bacteria
Microorganism
Bacteriocin
Inhibition/control
Reference
Bacillus cereus 8A
cerein 8A
Listeria monocytogenes
Bizani et al. (2008)
Enterococcus faecium L50
Enterocins L50A and
L50B
Listeria monocytogenes
Cintas et al. (1998)
Enterococcus faecalis S37
Enterocin S37
Listeria monocytogenes
Belguesmia et al. (2010)
Lactobacillus acidophilus
acidocin
Clostridium sporogenes
Leer et al. (1995)
Lactobacillus helveticus 481
Helveticin J
Lactobacillus bulgaricus 1489
Joerger and Klaenhummer (1986)
Lactobacillus plantarumC11
Plantaricins EF and JK
Staphylococcus spp.
Anderssen et al. (1998)
Lactobacillus plantarum ZJ008
Plantaricin ZJ008
Staphylococcus spp.
Zhu et al. (2014)
Lactobacillus plantarum ZJ5
Plantaricin ZJ5
Some Gram-positive bacteria
Song et al. (2014)
Lactobacillus sakei
Sakacin Q
Sakacin P
Lactobacillus sakei NCDO
2714 Lactobacillus
coryneformis
Diep et al. (2000)
Lactococcus lactis QU 4
Lactococcin Q
Lactococcus lactis
Zendo et al. (2006)
Lactococcus lactis
Nisin
Listeria monocytogenes
Beasley and aris (2004)
Leuconostoc mesenteroides
Mesentericin Y105
Listeria monocytogenes
Castano et al. (2005)
Pediococcus acidilactici
Pediocin AcH
Listeria monocytogenes
Biawes et al. (1991)
Pediococcus pentosaceous NCDC 273
pediocin PA-1
Listeria monocytogenes
Simha et al. (2012)
11
Bacteriocins-producing Gram-negative bacteria
Microorganism
Bacteriocin
Inhibition
Reference
Escherichia coli
Microcin J25
Salmonella newport
Vincent et al. (2004)
Escherichia coli
Microcin J25
Salmonella enterica
Rintoul et al. (2001)
Escherichia coli
Colicins
other strains of E. coli
Morales et al. (2015)
Escherichia fergusonii
Colicins
Escherichia coli
Símajs et al. (2002)
Klebsiella pneumoniae
microcin E492
Escherichia coli
Biologia et al. (1996)
Bacteriocins-producing Archaea
Microorganism
Bacteriocin
Inhibition
Reference
Halobacterium strain
GRB
Halocin G1
Sulfolobus spp.
Connor and Shand (2002)
12
Application of bacteriocin in
food product
13
Factors influencing the efficacy of bacteriocins
in food systems
Food-related factors
The food microbiota
The target bacteria
• –Food processing
conditions
• –Food storage temperature
• –Food pH, and bacteriocin
unstability to pH changes
• –Inactivation by food
enzymes
• –Interaction with food
additives/ingredients
• –Bacteriocin adsorption to
food components
• –Low solubility and
uneven distribution in the
food matrix
• –Limited stability of
bacteriocin during food
shelf life
•
•
•
•
• –Microbial load
• –Bacteriocin sensitivity
(Gram-type, genus,
species, strains)
• –Physiological stage
(growing, resting, starving
or viable but non culturable cells, stressed or
sub-lethally injured cells,
endospores ...)
• –Protection by physicochemical barriers
(microcolonies, biofilms,
slime)
• –Development of
resistance/adaptation
–Microbial load
–Microbial diversity
–Bacteriocin sensitivity
–Microbial interactions in
the food system
14
Galvez et al. (2007)
Cerein 8A product by B. cereus 8A to control
Listeria monocytogenes
.
day 0 (white bars)
day 3 (dashed bars)
day 5 (black bars).
Effect of cerein 8A to control Listeria monocytogenes in
pasteurized milk at 4 °C. Heat inactivated (Control) or
160 AU ml−1 cerein 8A (Cerein) were added to milk
samples before inoculation with L. monocytogenes.
15
Bizani et al, (2008)
Cerein
8A product
by B. cereus
control Listeria
Cerein
8A product
by8AB.to cereus
8A tomonocytogenes
control
Listeria monocytogenes
Control
cerein 8A 400 AU ml−1
Effect of cerein 8A to control Listeria monocytogenes
on surface of Minas-type cheese at 4 °C.
(Bizani et al, 2008)
16
Bacteriocin-producing Lactobacillus sakei C2 as
starter culture in fermented sausages
Trends of total enterobacteria
populations in the fermented sausages
Trends of L. monocytogenes
populations in the fermented sausages
Control
inoculated with5 log CFU/g of L. sakei C2.
inoculated with7 log CFU/g of L. sakei C2.
17
Gao et al. (2009)
Sensory evaluation of fermented sausages after the
fermentation of 38 days.
Sample
Inoculated with 7
Log CFU/g of L. sakei C2
Flavor
Texture
9.28 ± 0.62a 9.03 ± 0.23a
Overall acceptability
9.46 ± 0.59a
8.63 ± 0.52b 8.72 ± 0.41a
Inoculated with 5
Log CFU/g of L. sakei C2
8.87 ± 0.27b
8.15 ± 0.43c 8.97 ± 0.35a
8.47 ± 0.51b
The control
a, b, c
Means within the same column followed by different superscript letters
Differ significantly (p < 0.05).
Gao et al. (2009)
18
Pediocin 05-10 product by Pediococcus pentosaceus
Biocontrol of L. monocytogenes in pork ham by Pediocin 05-10 at 4 C.
Determination of the L. monocytogenes CFU was carried out every day
Batch I was pretreated with the bacteriocin and then incubated with L. monocytogenes
Batch II was pretreated with the inactive bacteriocin and then incubated with L. monocytogenes
19
Huang et al. (200
Effect of enterocin AS-48 on vegetative cells of B. coagulans
on tomato paste
37 oC
37 oC
B. coagulans CECT 12
(A–C)
B. coagulans CECT 561
(D–F)
22 oC
22 oC
4 oC
4 oC
 Enterocin AS-48 0µg/ml
 Entericin AS-48 3µg/ml
 Enterocin AS-48 6µg/ml
Effect of enterocin AS-48 on vegetative cells of B. coagulans CECT 12 (A–C)
and B. coagulans CECT 561 (D–F) inoculated on a commercial tomato paste
20
Lucas et al .(2006)
The potential for incorporating nisin into plastic films on
Cold-smoked salmon
4 oC
10 oC
5 × 102 CFU/cm2
4 oC
10 oC
5 × 105 CFU/cm2
21
Netoo et al .(2008)
Application of bacteriocins as part of
hurdle technology
22
Bacteriocins and high hydrostatic pressure (HHP)
Listeria monocytogenes
Room temperature , 22oC
7oC
Control of Listeria monocytogenes by enterocin
AS-48 (148 AU g1) alone or in combination with
HHP treatment in fuets during ripening and
storage at room temperature (A) or 7 C (B).
23
Non HHP, Non enterocin AS-48
Non HHP , enterocin AS-48
HHP , Non enterocin AS-48
HHP , enterocin AS-48
Ananou et al.(2010)
Salmonella enterica
Room temperature , 22oC
Control of S. enterica by enterocin AS-48 (148 AU g1)
alone or in combination with HHP treatment in fuet
sausages during ripening and storage at room
temperature (A) or 7C (B).
24
7oC
Non HHP, Non enterocin AS-48
Non HHP , enterocin AS-48
HHP , Non enterocin AS-48
HHP , enterocin AS-48
Ananou et al.(2010)
Conclusions
Additive
preservative
Bacteriocinproducing
starter
Hurdle
technology
25
Acknowledgement
Dr. Nitcha Chamroensaksri
26
27
References
Ananou,S, Garriga,M., Jofre,M., Aymerich,T., Galvez,A., Maqueda,M., Martinez-Bueno,M., Valdivia,E.(2010)
Combined effect of enterocin AS-48 and high hydrostatic pressure to control food-borne
pathogens inoculated in low acid fermented sausages. Meat Science ,84,594–600.
Bakkal, S., Robinson, S.M. and Riley, M.A. (2012) Bacteriocins of Aquatic Microorganisms and Their Potential
Applications in the Seafood Industry. In book: Health Environ Aquaculture, 303-328.
Bizani,D, Morrissy, A.C.J ,Dominguez, P.M.A , Brandelli,A. (2008) Inhibition of Listeria monocytogenes in dairy
products using the bacteriocin-like peptide cerein 8A . International Journal of Food Microbiology, 121,
229–233.
Gao,Y., Li,D., Liu ,X. (2014) Bacteriocin-producing Lactobacillus sakei C2 as starter culture in fermented
sausages .Food Control, 35 ,1- 6.
Huang,Y., Luo,Y., Zhai,Z., Zhang,H., Yang,Y., Tian,H.(2009) Characterization and application of an antiListeria bacteriocin produced by Pediococcus pentosaceus 05-10 isolated from Sichuan Pickle, a
traditionally fermented vegetable product from China. Food Control ,20,1030–1035.
28
Lucas,R., Grande,M.J., Abriouel,H., Maqueda,M., Omar,N.B., Valdivia,E., amero,M.M., Galvez,A. (2006)
Application of the broad-spectrum bacteriocin enterocin AS-48 to inhibit Bacillus coagulans in
canned fruit and vegetable foods. Food and Chemical Toxicology ,44,1774–1781
Neetoo,H., Ye.M., Chen,H., Joerger,R.D., Hicks,D.T. , Hoover,D.G. (2008) Use of nisin-coated plastic films to
control Listeria monocytogenes on vacuum-packaged cold-smoked salmon. International
Journal of Food Microbiology,122,8–15.
29