Production of Fermented foods and Beverages

1. Production of Fermented foods and Beverages Biotechnology

2. Food Biotechnology • Fermented food products • Alcoholic beverages • Production of organic acids • Production of amino acids • Food flavors • Single cell protein

3. Lactic acid bacteria (LAB) • A clade of Gram-positive, acid-tolerant, generally non-sporulating rods or cocci • Usually found in decomposing plants and lactic products, produce lactic acid as the major metabolic end-product of carbohydrate fermentation. • This trait has, links LAB with food fermentations, as acidification inhibits the growth of spoilage agents. • Bacteriocins are produced by several LAB strains and provide an additional hurdle for spoilage and pathogenic microorganisms. • Generally recognized as safe (GRAS) status, due to their ubiquitous appearance in food and their contribution to the healthy microflora of human mucosal surfaces.

4. Lactic acid bacteria • Common genera include • Lactobacillus • Leuconostoc • Pediococcus • Lactococcus • Streptococcus • Aerococcus • Carnobacterium • Enterococcus • Oenococcus • Sporolactobacillus • Tetragenococcus • Vagococcus • Weisella

5. Fermented food products • Milk Products • Fermented milk products have longer shelf life as compared to raw milk • Curd, buttermilk, yogurt and cheese require lactic bacteria • Butter and buttermilk • Made from cream/curd • From cream: soured by using Streptococcus cremorisor S. lactisfor the production of lactic acid • Leuconostoccremorisimparts characteristic flavor. • By churning of cream/curd, butter is produced, leaving buttermilk (liquid portion).

6. Yogurt • Fermentation of milk by • Lactobacillus bulgaricusand Streptococcus thermophilusat 40-45 ◦C. • After 4 hrs, sufficient acidity is generated, good to enhance shelf life. • Flavour is imparted by accumulation of lactic acid and acetaldehyde produced by L. bulgaricus. • Commercially produced from pasteurized skimmed milk with added milk powder.

7. Cheese, the milk protein • Cheese is a generic term for a diverse group of milk-based food products. • Cheese is produced throughout the world in wide-ranging flavors, textures, and forms. • Obtained by milk fermentation • Coagulated mixture of proteins including casein • Enzymetic process (rennin) • Cheese • Whey • Lactic bacteria are used • Renin may be obtained • From the calf stomach or • Produced by microorganisms

8. Cheese Varieties • Ripening of the cheese by • Bacteria • Mold • Soft cheese water content 50-80 % • Ripened by surface growth • Semi-hard 40-45% • Briefly cooked to lower the curd content • Hard cheese <40% • Ripened with inoculation of spores of moldPenicilliumroguefortii • Salting cheese with sodium chloride

9. Fermented Meats • A delicacy in some middle-east countries. • Fermentation of meat is carried out during curing by lactic bacteria and Pedicoccuscerevisiae. • Several types of salamis and sausages are produced by fermentation • Gives flavour • Preserves food

10. Leavening bread • The dough is fermented in the presence of sugar and yeast. • Saccharomycescerevisiae, the Baker’s yeast is mostly used in leavening process. • Production of Baker’s yeast • Normally grown at 30◦C on molasses which is • Having low conc. of sugar (0.5-1.5%) • Rich in mineral salts • pH about 4.5 • During fermentation, sugar content of molasses is meticulously controlled as higher conc. is deterrent to respiratory enzymes. • Yeast is collected through centrifugation.

11. Alcoholic Beverages • Vinegar is an alcoholic liquid that has been allowed to sour. • Used to flavor and preserve foods. • Ingredient in salad dressings and marinades. • Used as a cleaning agent. • Two successive fermentations of grape juice, raisins or malt • S. cerevisiaeanerobically converts carbs to alcohol • Oxidative transformation of alcohol to acetic acid by Acetobacterand Gluconobacter. • Starting material includes • Citrus fruits • Apple • Pear • Vegetables like potatoes • Malted cereals • Sugary syrups such as molasses, honey etc.

12. Production of organic acids • Acetic acid • Citric acid • Gluconic acid • Lactic acid • Gibberallic acid

13. Citric acid production • Citric acid is a weak organic acid. • It is a natural preservative/conservative and is also used to add an acidic, or sour, taste to foods and soft drinks. • Aspergillusnigeris used for commercial production • Various strains of A. niger are used in the industrial preparation of citric acid and gluconic acid and have been assessed as acceptable for daily intake by the World Health Organisation.

14. Citric acid Uses • Cleaning • Citric acid's ability to chelate metals makes it useful in soaps and laundry detergents. • By chelating the metals in hard water, it lets these cleaners produce foam and work better without need for water softening. • Citric acid is the active ingredient in some bathroom and kitchen cleaning solutions. • A solution with a 6% concentration of citric acid will remove hard water stains from glass without scrubbing. In industry, it is used to dissolve rust from steel. • Citric acid can be used in shampoo to wash out wax and coloring from the hair.

15. Cosmetics and pharmaceuticals • Citric acid is commonly used as a buffer to increase the solubility of brown heroin. • Single-use citric acid sachets have been used as an inducement to get heroin users to exchange their dirty needles for clean needles in an attempt to decrease the spread of AIDS and hepatitis. • Citric acid is used as one of the active ingredients in production of antivirals. Dyeing • Citric acid can be used in food coloring to balance the pH level of a normally basic dye. • Citric acid is used as an odorless alternative to white vinegar for home dyeing with acid dyes. Foods, beverages, and personal care • As a food additive, citric acid is used as a flavoring and preservative in food and beverages, especially soft drinks. • Citric acid can be added to e.g. ice cream as an emulsifying agent to keep fats from separating, to caramels to prevent sucrose crystallization, or to recipes in place of fresh lemon juice. • Citric acid is used with sodium bicarbonate in a wide range of effervescent formulae, both for ingestion (e.g., powders and tablets) and for personal care.

16. Lactic Acid • Used as a preservative and also in leather and textile industries • Microbes involved in lactic acid production are • Lactobacillus delbrueckii • Streptococcus and Leuconostoc species. • Typical fermentation medium contains • Gluscose 10-15% • Calciun carbonate 10% • Ammonium phosphate • Trace elemnets • Fermentation at 40-50 ◦C, pH 5.5-6.5 for 5-7 days

18. Production of Amino Acids • Amino acids are consumed in a variety of markets. • The largest by volume is the food flavoring industry. • Monosodium Glutamate, alanine, aspartate, arginine are all used to improve the flavor of food. • L-lysine is directly produced from carbs by using Corynebacteriumglutamcum, an auxotroph • MSG is produced by Arthrobacter, Corynebacteriumand Brevibacterium

19. Food Flavours • All kinds of preserved foods, such as soft drinks, milk products and beverages are flavoured suitably. • The flavouring compounds are mostly purine nucleotides, including inosinic acid (IMP), and GMP. • Yeast is commercial source of flavouring compounds • Bacillus subtilisalso enzymetically produces purine nucleotides through fermentation

20. Single-cell protein (SCP) • SCP refers to the dried microbial cells or total protein extracted from pure microbial cell culture (monoculture – Algae, bacteria, filamentous fungi, yeasts, etc…), which can be used as food supplement to humans (Food Grade) or animals (Feed grade). • SCP contains high protein content (60 – 80% of dry cell weight), fats, carbohydrates, nucleic acids, vitamins, and minerals. • It is also rich in essential amino acids such as Lys and Met.

21. SCP from Algal sources • Economical as algae utilize solar energy for growth. • Chlorella and Spirulina(blue-green algae). • Food from these sources contain 65% protein, 20% carbohydrates and remaining 15% include lipids, fibre, pigments etc. SCP from Bacteria • Cheap SCP is obtained from bacteria growing on cheap agricultural and mineral wastes • CellulomonasandAlcaligenesare mostly used Fungi as source of SCP • Saccharomyces, Candida utilis and Torulopsiscan be grown on wastes from paper industry and forestry • Rhodosporiumsphaeroidesproduce a cell mass that contains more than 50% lipids.

22. Advantages of using microorganisms for SCP production • Protein synthesis is much more rapid than higher living systems. • Microbes have short generation time. • Easily modifiable genetically for determining the amino acid composition. • Microbes have high protein content (7.12g protein Nitrogen/100g dry weight). • Microbes can be grown on media containing cheap sources of C and N. • Easy regulation of environmental factors for efficient yield.

23. Disadvantages • Possibility of contamination with pathological organisms in SCP. • Association of carcinogenic and other toxic substances with SCP is often observed. • Digestion of microbial cells is rather slow, and is frequently associated with indigestion and allergy reactions. • Food grade production of SCP is more expensive than othersources of proteins, as it depends on the raw materials. • SCP for human consumption is 10-12 times more expensive thanSCP for animal feed.