Lectures 15 Notes
Lectures 15 Notes
WHAT IS IMPORTANT?
▪ Why fermentation?
▪ Malolactic fermentation.
▪ Know different fermented food products, how they are produced (processes and
starter cultures used), what or which organisms cause their spoilage.
INTRODUCTION
• Discovery: before mankind knew of microbes, but witnessed effects of their activity.
• Foods are fermented for improvement of shelf life and safety of product.
• Produced by actions of LAB and fungi, mainly yeasts and to a lesser extent, moulds.
• LAB and facultative yeasts grow under anaerobic conditions and therefore normally occur together in
fermented foods.
• Either act together to produce a product, or one group plays a role as spoilage organism.
YEASTS
• Single celled, true fungi, reproducing either by budding or fission. A few can develop
the filamentous structure of a typical mould.
• Natural habitat: nutrient rich environments e.g. nectarines of plants, plant exudates,
decaying fruits and the body fluids of animals.
• Only a small number involved in food fermentation though large diversity of yeasts
exists.
• Zygosaccharomyces rouxii: Associated with the fermentation of plant products in which the
addition of salt is an integral part of the process.
• Hansenula anomala and Debaryomyces hansenii can grow in concentrated salt solutions. D.
hansenii: Isolated from brined meat products and fermented sausages.
YEASTS
• Pichia guilliermondii and Saccharomycopsis fibuligera: Isolated from tropical
fermented products.
• Causes spoilage of products stored in plastic packs and glass bottles causing
explosion of the container.
• Facultative heterofermenters (Lb. plantarum, Lb. casei and Lb. sake) ferment hexoses
via the EMP pathway to lactate but have an inducible phosphoketolase which allows them
to ferment pentoses to lactate and acetate.
• Obligate heterofermenters (Lb. brevis, Lb. fermentum, Lb. kefir) use the
phosphoketolase pathway for hexose fermentation.
• Leuconostoc : Produce only D-lactate and are unable to produce ammonia from arginine.
• Pediococcus include species such as P. pentosaceus, and P. halophilus reclassified in the genus of
its own, Tetragenococcus halophilus.
• Bifidobacterium are included by some authors among LAB though different both
phylogenetically and biochemically.
LAB ACTIVITIES IN FOOD
• LAB inhibits other microbes thereby improving keeping quality and safety of food products.
Bacteriocins
• Bacterial proteins active against species of closely related to producing strain.
• Natural food preservative produced by food grade microbes.
• Nisin: Produced by certain strains of Lactococcus lactis. Have broad spectrum against G+. G- whose membranes have
been damaged by thermal shock are also sensitive. Bacterial spores are sensitive and nisin is used to inhibit their
outgrowth in heat processed products.
Hydrogen peroxide
• Produced by LAB in the presence of O2. It potentiates the lactoperoxidase antimicrobial system which catalyse
oxidation of thiocyanate by H2O2 to produce hypothiocyanate which kills G- and inhibits G+.
Ethanol
• Produced by heterofermentative LAB and contributes to the inhibition of competitors.
• Lactose intolerance
• Individuals lacking the enzyme β-galactosidase which hydrolyses lactose into glucose and galactose.
• The lactose passes to small intestine undigested and is then attacked by resident population of lactose
fermenting microbes producing abdominal discomfort, flatulence and diarrhoea.
• Adverse effects less severe with consumption of viable starter cultures in yoghurt.
• The starter cultures use their β-galactosidase activity to assist in hydrolysis of lactose.
• Inhibition of pathogens
• LAB colonising the gut mask the potential attachment sites in the gut for pathogenic microorganisms.
• They also have antitumor activity where the activity of enzymes such as β-glucuronidase, azoreductase and
nitroreductase (convert procarcinogens to carcinogens)in faeces are reduced.
MALOLACTIC FERMENTATION
• LAB decarboxylate a strong acid, L-malic acid, to produce a weak acid, L-lactate.
Encouraged in wines from cool regions with high acidity.
• Provides bacteriological stability to the bottled product and, modify and improve
the body and flavour of wine.
• Encouraged naturally by leaving the new wine on yeast sediment for longer than
usual.
Cool : Ensures that starter cultures are not killed by high temperature
Incubate for 4h : Starter bacteria ferment lactose to lactic acid reducing the pH from 6.3-6.6 to 3.8-4.2
Cool(15-20oC)
• However, yoghurts containing fruits may have up to 100 yeasts and still remain of
satisfactory quality
• Milk homogenized and cooled to 37-40oC before addition of 2-5% starter (Lb. acidophilus).
• Drawbacks:
• Unlike yoghurt, it has a sour acidic taste.
• Shelf life of a week at 5oC due to poor survival of Lb. acidophilus in acidic conditions.
• Non fermented sweet acidophilus milk is produced by addition of starter culture to pasteurized milk
without incubation.
Bioyoghurts
• Produced in an attempt to combine qualities of acidophilus milk and yoghurt.
• Contain organisms able to colonize gut such as Lb. acidophilus and Bifidobacterium spp., and S. thermophilus
to provide characteristic yoghurt flavor.
Kefir and koumiss
• Both produced by mixed LAB and alcoholic fermentation.
• Kefir
• Microflora added as kefir grains with smooth outer surface containing LAB and inside
rough surface containing a mixture of yeast and LAB.
• Flavour compounds are ethanol, lactic acid, carbon dioxide, acetaldehyde and diacetyl.
• Koumiss
• Fizzy, greyish white drink produced traditionally from mare’s milk (modified cow’s milk).
• Produced by activities of Lb. delbrueckii subsp. bulgaricus and a number of lactose
fermenting yeasts.
KEFIR PRODUCTION
Homogenize milk
Cool to 22oC
• Practised on relatively small scale accounting for rich diversity (78 types of blue and 36
Camamberts). Table 9.7 A & M
• Most popular is cheddar cheese with a smooth texture and good keeping qualities.
• Keeping qualities vary with type but are always better than those of milk.
Cool milk to fermentation temperature: 29-31oC for cheddar, Stilton, Leister, Wensleydale
Addition of starter culture as concentrated cultures (WHY??) at 106-107 cfu/ml
Convert lactose to lactic acid{shelf life, safety,flavour}; Weakens casein and release
Mesophillic: Lactococcus lactis
calcium from it : aids chymosin action; Moisture expulsion and curd shrinkage
Thermophillic: Lb. helveticus, Lb. casei, Lb. lactis, Lb. bulgaricus, S. thermophilus
Incubate for 45 min: Starter multiply to 108-109 cfu/g and produce 0.6-0.7% acidity
Curdling: Add rennet & incubate for 30-45 min Coagulates milk: chymosin/rennin cleaves k-casein forming
cross links between micelles to form network
• Problems of bacteriophages
• Slows or stops acidification: Financial losses to producer; Increased risk of pathogens
• Propagation of starters in phage inhibitory media containing phosphate salts to chelate Mg2+
and Ca2+ ions
• Intrinsic methods of LAB resistance to phage infection
• Restriction/modification of non-host DNA