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BIOPRESERVATION
K R MICRO NOTES 1
INTRODUCTION
 Contamination and spoilage by pathogens is a great
concern.
 Many chemical preservatives and physical treatments are
being used but they have many drawbacks.
 Therefore, biopreservation is a desirable alternative.
K R MICRO NOTES 2
BIOPRESERVATION
 Extension of shelf life and food safety by the use of
natural or controlled micro biota and/or their antimicrobial
compounds.
 Beneficial bacteria or the fermentation products used in
biopreservation.
 Various modes of action are: organic acid production, a
wide variety of small inhibitory molecules including
hydrogen peroxide, etc.
K R MICRO NOTES 3
BIOPRESERVATIVE AGENTS
Lactic acid
bacteria:
• Have
antagonistic
properties.
• Compete for
nutrients, their
metabolites,
hydrogen
peroxide, and
peptide
bacteriocins.
Yeast:
• competition for
nutrients,
production and
tolerance of
high
concentrations
of ethanol.
• synthesis of a
large class of
antimicrobial
compounds.
Bacteriophages
• viruses which
infect bacteria.
• Controls certain
bacterial
pathogens.
K R MICRO NOTES 4
LACTIC ACID BACTERIA
 Include the genera Lactococcus, Streptococcus,
Lactobacillus, Pediococcus, Leuconostoc, Enterococcus,
etc.
 Preservation of the nutritional qualities of the raw material
through extended shelf life and the inhibition of spoilage
and pathogenic bacteria.
K R MICRO NOTES 5
LAB BACTERIOCINS
Class I:
• Comprises the lantibiotics.
• Small peptides differentiated from others by their
content in dehydro amino acids and thioether
amino acids. E.g nisin
Class II:
• Comprises thermostable non-lantibiotic linear
peptides.
• Divided into three subclasses : the pediocin-like
bacteriocins (class II.1), the lack of leader peptide
(class II.2), or neither of the above traits (class
II.3).
K R MICRO NOTES 6
Class III:
• Large heat-labile bacteriocins that
encompass many bacteriolytic extracellular
enzymes that may mimic the physiological
activities of bacteriocins.
• Examples :helveticin J of L. helveticus.
Class IV:
• New class created to include circular
antibacterial peptide.
• Distinguishing characteristic: existence of
head-to-tail peptide chain ligation.
K R MICRO NOTES 7
REQUIREMENTS AND REGULATORY STATUS FOR
BACTERIOCINS
 GRAS status.
 Should have a broad spectrum.
 Thermostability.
 Beneficial effects and improved safety.
 No adverse effect on quality and flavour.
K R MICRO NOTES 8
APPLICATIONS OF BACTERIOCIN-PRODUCING
LAB IN FOOD
 Inoculation of food with LAB where bacteriocins are
produced in situ.
 Use of food previously fermented with the
bacteriocin-producing strains as an ingredient in the
food processing (NisaplinTM, MicrogardTM, AltaTM
2341).
 Addition of purified or semipurified bacteriocins.
K R MICRO NOTES 9
BACILLUS AS BIOPRESERVATIVE
Subtilin from B.
subtilis
Megacin from B.
megaterium
Thermacin from B.
stearothermophilus.
K R MICRO NOTES 10
 Antimicrobial action against Gram positive and Gram
negative bacteria, as well as fungi.
 Safe for humans and would be no more of a risk than
lactic acid bacteria.
 The physiology/genetics of Bacillus are well understood.
 Inhibitory activity at alkaline, acidic condition or after
thermal processing and would be suitable for food
processing.
K R MICRO NOTES 11
MEAT BIOPRESERVATION
 Extensively studied in fermented meat products and ready
to eat meat products.
 Competitive enhancement strategies include:
 A natural 'green' method
competitive
exclusion
prebiotic
probiotic
K R MICRO NOTES 12
SEAFOOD BIOPRESERVATION
 Adding antimicrobials or by increasing the acidity of the
fish muscle.
 Less documented studies.
 Selection of potential protective bacteria in seafood
products is challenging.
 LAB remains the category that offers the highest
potential.
K R MICRO NOTES 13
APPLICATIONS IN VEGETABLE PRODUCTS
 Nisin in tinned vegetables and fruit juices.
 Pediocin in salad and fruit juice.
 Enterocin AS-48 against B. cereus in rice and vegetables.
 In fruit juices against other pathogens such as E. coli
O157:H7, S. aureus, and the spoilage bacterium
Alicyclobacillus acidoterrestris
K R MICRO NOTES 14
APPLICATION OF BACTERIOCINS IN DAIRY PRODUCTS
Nisin and/or nisin-producing
strains against Clostridium
butulinum in cheese and
against L. monocytogenes in
cheeses such as Camembert
, Ricotta , and Manchego .
Pediocin AcH in milk and
Cheddar and Munster
cheeses against L.
monocytogenes, S. aureus,
and E. coli O157:H7.
Lacticin 3147 against
undesirable LAB, L.
monocytogenes and B.
cereus in Cheddar, Cottage
cheese and yogurt .
Enterocin AS-48 against B.
cereus, S. aureus and L.
monocytogenes in milk.
K R MICRO NOTES 15
BACTERIOPHAGES
 This specificity has allowed for directed targeting
and killing of pathogenic bacteria using phages in
what is referred to as phage therapy.
K R MICRO NOTES 16
MODE OF ACTION
K R MICRO NOTES 17
K R MICRO NOTES 18
SAFETY
 LAB and their bacteriocins have been consumed
unintentionally for ages, laying down a long history of safe
use.
 Factors: antimicrobial spectrum of inhibition, bactericidal
mode of action, relative tolerance to processing conditions
and the lack of toxicity.
 The evaluation done in meat by USDA.
K R MICRO NOTES 19

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Biopreservation .pptx

  • 2. INTRODUCTION  Contamination and spoilage by pathogens is a great concern.  Many chemical preservatives and physical treatments are being used but they have many drawbacks.  Therefore, biopreservation is a desirable alternative. K R MICRO NOTES 2
  • 3. BIOPRESERVATION  Extension of shelf life and food safety by the use of natural or controlled micro biota and/or their antimicrobial compounds.  Beneficial bacteria or the fermentation products used in biopreservation.  Various modes of action are: organic acid production, a wide variety of small inhibitory molecules including hydrogen peroxide, etc. K R MICRO NOTES 3
  • 4. BIOPRESERVATIVE AGENTS Lactic acid bacteria: • Have antagonistic properties. • Compete for nutrients, their metabolites, hydrogen peroxide, and peptide bacteriocins. Yeast: • competition for nutrients, production and tolerance of high concentrations of ethanol. • synthesis of a large class of antimicrobial compounds. Bacteriophages • viruses which infect bacteria. • Controls certain bacterial pathogens. K R MICRO NOTES 4
  • 5. LACTIC ACID BACTERIA  Include the genera Lactococcus, Streptococcus, Lactobacillus, Pediococcus, Leuconostoc, Enterococcus, etc.  Preservation of the nutritional qualities of the raw material through extended shelf life and the inhibition of spoilage and pathogenic bacteria. K R MICRO NOTES 5
  • 6. LAB BACTERIOCINS Class I: • Comprises the lantibiotics. • Small peptides differentiated from others by their content in dehydro amino acids and thioether amino acids. E.g nisin Class II: • Comprises thermostable non-lantibiotic linear peptides. • Divided into three subclasses : the pediocin-like bacteriocins (class II.1), the lack of leader peptide (class II.2), or neither of the above traits (class II.3). K R MICRO NOTES 6
  • 7. Class III: • Large heat-labile bacteriocins that encompass many bacteriolytic extracellular enzymes that may mimic the physiological activities of bacteriocins. • Examples :helveticin J of L. helveticus. Class IV: • New class created to include circular antibacterial peptide. • Distinguishing characteristic: existence of head-to-tail peptide chain ligation. K R MICRO NOTES 7
  • 8. REQUIREMENTS AND REGULATORY STATUS FOR BACTERIOCINS  GRAS status.  Should have a broad spectrum.  Thermostability.  Beneficial effects and improved safety.  No adverse effect on quality and flavour. K R MICRO NOTES 8
  • 9. APPLICATIONS OF BACTERIOCIN-PRODUCING LAB IN FOOD  Inoculation of food with LAB where bacteriocins are produced in situ.  Use of food previously fermented with the bacteriocin-producing strains as an ingredient in the food processing (NisaplinTM, MicrogardTM, AltaTM 2341).  Addition of purified or semipurified bacteriocins. K R MICRO NOTES 9
  • 10. BACILLUS AS BIOPRESERVATIVE Subtilin from B. subtilis Megacin from B. megaterium Thermacin from B. stearothermophilus. K R MICRO NOTES 10
  • 11.  Antimicrobial action against Gram positive and Gram negative bacteria, as well as fungi.  Safe for humans and would be no more of a risk than lactic acid bacteria.  The physiology/genetics of Bacillus are well understood.  Inhibitory activity at alkaline, acidic condition or after thermal processing and would be suitable for food processing. K R MICRO NOTES 11
  • 12. MEAT BIOPRESERVATION  Extensively studied in fermented meat products and ready to eat meat products.  Competitive enhancement strategies include:  A natural 'green' method competitive exclusion prebiotic probiotic K R MICRO NOTES 12
  • 13. SEAFOOD BIOPRESERVATION  Adding antimicrobials or by increasing the acidity of the fish muscle.  Less documented studies.  Selection of potential protective bacteria in seafood products is challenging.  LAB remains the category that offers the highest potential. K R MICRO NOTES 13
  • 14. APPLICATIONS IN VEGETABLE PRODUCTS  Nisin in tinned vegetables and fruit juices.  Pediocin in salad and fruit juice.  Enterocin AS-48 against B. cereus in rice and vegetables.  In fruit juices against other pathogens such as E. coli O157:H7, S. aureus, and the spoilage bacterium Alicyclobacillus acidoterrestris K R MICRO NOTES 14
  • 15. APPLICATION OF BACTERIOCINS IN DAIRY PRODUCTS Nisin and/or nisin-producing strains against Clostridium butulinum in cheese and against L. monocytogenes in cheeses such as Camembert , Ricotta , and Manchego . Pediocin AcH in milk and Cheddar and Munster cheeses against L. monocytogenes, S. aureus, and E. coli O157:H7. Lacticin 3147 against undesirable LAB, L. monocytogenes and B. cereus in Cheddar, Cottage cheese and yogurt . Enterocin AS-48 against B. cereus, S. aureus and L. monocytogenes in milk. K R MICRO NOTES 15
  • 16. BACTERIOPHAGES  This specificity has allowed for directed targeting and killing of pathogenic bacteria using phages in what is referred to as phage therapy. K R MICRO NOTES 16
  • 17. MODE OF ACTION K R MICRO NOTES 17
  • 18. K R MICRO NOTES 18
  • 19. SAFETY  LAB and their bacteriocins have been consumed unintentionally for ages, laying down a long history of safe use.  Factors: antimicrobial spectrum of inhibition, bactericidal mode of action, relative tolerance to processing conditions and the lack of toxicity.  The evaluation done in meat by USDA. K R MICRO NOTES 19