BACTERIOCIN

antibacterial substance,such as colicin, produced by a strain of certain bacteria and harmful to another strain within the family. Bacteriocins are proteinaceous toxins produced by bacteria to inhibit the growth of similar or closely related bacterial strains.  Bacteriocins were first discovered by A.Gratia in 1925.
An

Bacteriocins produced LAB is suitable for food preservation

 Are not active and nontoxic on eukaryotic cells.  Become inactivated by digestive proteases ,having little influences on the gut microbiota.  Are generally recognised as safe substances.  Are usually ph and heat tolerant.  They have a antibacterial spectrum,against many food borne pathogenic and spoilage bacteria.  They show a bactericidal mode of action,usually acting on the bacterial cytoplasmic membrane.  No cross resistance with antibiotics.  Their genetic determinants are usually plasmid encoded, facilitating genetic manipulation.

Factors influencing the efficacy of bacteriocins in food systems

 Food processing conditions.  Food storage temperature.  Food Ph ,and bacteriocin unstability to ph changes.  Inactivation of food enzymes.  Interaction with food additives and ingredients.  Bacteriocin adsorption to food components.  Low solubility and uneven distribution in the food

matrix.  Limited stability of bacteriocin during food shelf life.

Application of bacteriocin in food preservation

 An extended shelf life of foods.  Provide extra production during temperature abuse conditions.  Decrease the risk for transmission of foodborne pathogens through the foodchain.  Ameliorate the economic losses due to food spoilage.  reduce the application of chemical preservatives.  Permit the application of less severe heat treatments without compromising food safety.  Permit the marketing of novel foods.  They may serve to satisfy industrial and consumer demands.

CONT ..
 Microbial load.  Microbial diversitiy.  bacteriocin sensitivity (gram type,genus ,species

strains.  Physiological state.(growing resting starving or viable but non culturable cells, stressed or sub- lethally injured cells,endospores.  Protection by physico chemical barriers.  development of resistance /adaption

Effectiveness of bacteriocins in food systems

 Factors negatively affecting production.  It includes inadequate physical conditions and chemical composition         

of food. (ph, temperature, nutrients,etc. ) Spontaneous loss in production capacity. Inactivation by phage of the producing strain. Antogonism effect of other microorganisms in foods. The effectiveness of bacteriocin activity in food is negatively affected by resistance development of pathogens to the bacteriocin. Inadequate environmental conditions for the biological activity. Higher retention of the bacteriocin molecules by food system components. Inactivation by other additives. Slower diffusion and solubility and or irregular distribution of bacteriocin molecules in the meat matrix.

Requirements and regulatory status for bacteriocin

 The producing strain should preferably have GRAS

status.  Thermostability.  Beneficial effects and improved safety.  No adverse effect on quality and flavour.  The bacteriocin should have a broad spectrum of inhibition.

Application of bacteriocin producing LAB in food

 Inoculation of food with LAB where bacteriocins are

produced in insitu.  Use of food previously fermented with the bacteriocin producing strains as an ingredient in the food processing.  Addition of purified or semipurified bacteriocins.  The potential of bacteriocin producing LAB and their bacteriocins,especially lactococci, pediococci,lactobacilli, and enterococci

Bacteriocins and hurdle technology

 The concept of hurdle technology to apply in the food

survival of microorganisms greatly decreased when they were confronted with antimicrobial factors.  After exposure of a bacterial population to a single antimicrobial factor .  It is a heterogenous response,depending on the intensity of treatment.  A fraction of the population may receive a lethal dose of the antimicrobial factor ,leading to cell death.

Cont .
 The remaining fraction may survive due to several

reasons.  Receiving a sub lethal dose.  Showing an increased resistance because of its physiological state.  Cells naturally resistant to the antimicrobial agent.

Application of bacteriocins as part of hurdle technology

 Bacteriocins can be used purposefully in combination

with selected hurdles to increase microbial inactivation.  The combination of hurdles to be applied will depend greatly on the type of food and its microbial compositions.  Use of bacteriocins combined with other preservation methods to create a series of hurdles during the manufacturing process to reduce food spoilage by microorganisms.

CONTD .
 The application of chemical preservatives,physical

methods (pulsed electric field,HHP,vaccum or modified atmospheric packaging).  Which increase the permeability of cell membrane.  Positively affects the activity of many bacteriocin.

Combination of bacteriocins with chemical substances and natural antimicrobials

presence of Nacl enhanced the antimicrobial action of bacteriocins such as nisin,leucocin F10,enterocin.  Sodium chloride also decreased the antilisterial activity of acidocin,pediocin, curvocin.  The protective effect of Nacl may be due to interference with ionic concentrations between bacteriocin molecules and charged groupes involved in bacteriocin binding to target cells.  Sodium chloride may also induce conformational changes of bacteriocins or changes of the cell envelope of the target organisms.
 The

Bacteriocins and heat treatments

It can be used to reduce the intensity of hear without compromising microbial inactivation.  Nisin and heat act synergistically against L. plantarum and L. monocytogenes reducing the heat resistance of L. monocytogenes in milk and in cold-pack lobster.  Nisin-resistant L. monocytogenes cells grown in the presence of nisin were more sensitive heat at 55C than wild-type cells.  The efficacy of enterocin was higher on S. aureus cells sub-lethally injured by heat due to the lower concentration of remaining viable cells and to the cell damage induced by the heat treatment.

Con
I n so prov d n dd on pro on dur ng ood s or g g ns pro r on o ndospor s surv v ng h r m ns  In ns y o h r m n s g ns b r ndospor s n b ow r d n omb n on w h n s n s w s w h n ro n  ub- h h o s ns z Gr m-n g v b r o s v r b r o ns su h s n s n or p d o n, n ro n  Ex nd ng h r sp rum o on  H gh s s ns z on w s omb n d r m n s o b r o ns, h nd h ng g n s

B t

ns nd m d f d tm sph

p k g ng

 Modified atmos ere packaging is sed in the food industr to prolong the shelf life of perishable food products  MAP is defined as the enclosure of food products in gasbarrier materials, in which the gaseous environment has been changed  It is based on retardation of intrinsic food changes and inhibition of spoilage microbiota.  In a modified atmosphere, the dissolved O2, will determine growth inhibition of microorganisms.  Gram-negative bacteria are generally more sensitive to O2, while lactic acid bacteria are much more resistant.

Cont..
 Sin e Gram-negative acteria are usually not sensitive to bacterio ins, MAP and bacterio ins are therefore two omplementary hurdles of a vantage to food spoilage. 

Ba t

sa

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l t

f l s

 It is a non-thermal ro ess microbial inactivation is achieve y a lication of high-voltage ulses between a set of electrodes.  The effects of PEF resemble acterial electroporation, but the higher intensity of this treatment causes were amage to the acterial cell membrane.  This technology an only be a lie to food produ ts.  Bacterio ins act on the acterial cytoplasmic membrane, the combine a lication of acterio ins.  PEF is expecte to in rease acterici al effects.  PEF treatments, su h as sub-lethally in ure cells or bacterial endospores.

Cont..
 PEF oul also be a lie to extend the antimicrobial spectrum of acterio ins, sin e PEF disrupts the acterial outer membrane allowing bacterio in molecules to reach the acterial cytoplasmic membrane target.  Factors relate to the PEF treatment the food microbial loa , omposition and physiological stage, the a e acterio in, and other environmental factors.

Ba t

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ssu

 High hydrostatic ressure is an innovative food pro essing and preservation method that causes in ury and killing of microbial cells.  During pressurization, the isruption of H-bonds, ionic onds and hydrophobic interactions of the macromolecules a versely affects their stru tures and fun tions.  The sub-lethal amage is initiate y membrane hase transitions affecting mainly ATP-generating and transport roteins.  Cell eath ause y HHP in reases with pressure and so does the synergism with bacterio ins.  Most acterio ins act on the acterial cytoplasmic membrane it can be hypothesize that the observe synergy between bacterio ins and HHP results from umulative amage to this stru ture.

Cont
 HHP treatments indicating that cell eath o urs as a consequen e of multi le events or umulative cell amage.  Sub-lethally in ure vegetative, ells surviving HHP treatment may develop pressure resistan e.  The in rease cell amage cause y ombine treatments of HHP and bacterio ins oul revent the tailing effect, provi ing an a itional hurdle against selection of ressure-resistant vegetative cells.

Effect of pulsed electric fields in combination with bacteriocins on microbial populations

Application of bacteriocins as part of hurdle technology

Cont
 The bactericidal effect of HHP increases along with the temperature,which determines the different modalities of treatment.e.g cold HHP pasteurisation.  The food ph is also an influencing factor,and bacteria are usually more resistant to HHP in low acid foods.  Bacteriocins could improve the efficacy of heat treatments in foods,compensating for the required increase in pressure or temperature.  Bacteriocins are generally inactive on Gram negative bacteria  Gram negative bacteria through outer membrane damage ,increasing the possibilities for application of bacteriocins in food preservation.

Cont .
 HHP also induces a more persistent sensitisation of

Gram negativebacteria to small diffusible antimicrobial molecules bacterial endospores which may act synergestically with other hurdles.  Bacterial endospores are resistant to HHP treatments applied to foods,HHP treatments can induce endospore germination.  Addition of bacteriocins against surviving endospores could improve the safety and shelf life of HHP processed foods.

Cont
 Reduction of nitrite content by addition of

bacteriocins may be benefical in the food industry.  The combinations of nisin and nitrite delayed botulinal toxin formation in meat systems.  It showed increased activity on clostridial endospores and outgrowth L.mesenteroides and L.monocytogenes.  Addition of nitrite also inreased the anti-listeria activity.

Cont
 Organic acid and their salts can potentiate the activity

of bacteriocins greatly,while acidifiction enhances the antibacterial activity of both organic acids and bacteriocins.  The increase in net charge of bacteriocins at low ph might facilitate translocation of bacteriocin molecules through the cell wall.  The solubility of bacteriocins may also increase at lower ph ,facilitating diffusion of bacteriocin molecules.

Cont .
 Chelating agents permeate the outer membrane of

gram negative bacteria by extracting ca and mg cations that stabilize lipopolysacharide of this structure,allowing bacteriocins to reach the cytoplasmic membrane.  The enhanced effect of chelators such as EDTA,disodiumphosphate,trisodium phosphate,hexametaphosphate or citrate and bacteriocins against gram negative bacteria.  Chelating agents can also enhance the activity of bacteriocins on gram positive bacteria.

Conclusion
 The use of bacteriocins and bacteriocin producing

strains of LAB are generally recognized as safe organisms and their antimicrobial products as biopreservatives.  Understanding of the influences that environmental factors have on the implantation and survival of bacteriocinogenic strains and the activity of their bacteriocins.  Quantitatively estimate their efficiency for future applications in food model systems and application of these biopreservatives.