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Fermentation: Fermentation may be defined from different perspective such asIn Biochemistry it may defined as a process that is important

in anaerobic conditions when there is no oxidative phosphorylation to maintain the production of ATP (Adenosine triphosphate) by glycolysis. During fermentation pyruvate is metabolised to various different compounds. Homolactic fermentation is the production of lactic acid from pyruvate; alcoholic fermentation is the conversion of pyruvate into ethanol and carbon dioxide; and heterolactic fermentation is the production of lactic acid as well as other acids and alcohols. In case of Food Fermentation typically refers to the conversion of sugar to alcohol using yeast under anaerobic conditions. A more general definition of fermentation is the chemical conversion of carbohydrates into alcohols or acids. In case of Biotechnology fermentation means any process by which microorganisms are grown in large quantities to produce any type of useful materials. Pharmaceuticals and the biotechnology industry: There are 5 major groups of commercially important fermentation: 1. Microbial cells or biomass as the product, e.g. bakers yeast, lactobacillus, etc. 1. Microbial enzymes: catalase, amylase, protease, pectinase, glucose isomerase, cellulase, hemicellulase, lipase, lactase, streptokinase, etc. 2. Microbial metabolites : 1. Primary metabolites ethanol, citric acid, glutamic acid, lysine, vitamins, polysaccharides etc. 2. Secondary metabolites: all antibiotic fermentation 3. Recombinant products: insulin, HBV, interferon, GCSF, streptokinase 4. Biotransformation: phenyl acetyl carbinol, steroid biotransformation, etc. Requirements for Fermentation: For carrying out the fermentation process various goods are required which include1. Fermenters: The heart of fermentation process is fermenter. A working definitions of a fermenter is a container in which, is maintained an environment favorable to the operation of a desired biological process. The common features of typical fermenters are as follows

They should be strong enough to withstand the pressure exerted by large volume of the medium. The material used for the construction of fermenter should not be corroted by the fermentation product and it should not yield toxic ions to the medium. The fermenter should have provision for the control or prevention of the growth of contaminating microorganisms. This is a must because the industrial fermentation requires pure cultures. If aerobic organisms are used in the process, there should be provision for rapid incorporation of sterile air into the medium so that the oxygen is immediately dissolved in the medium and available to the microorganisms. The carbon dioxide produced by the microorganisms should be removed from the medium and provision should be made for this. Certain kind of stirring is necessary to mix the organisms with medium and to make nutrients and oxygen available to individual microbe. A system should be available for detection of PH of the culture medium and also for its adjustment.

2. Microorganisms: Many species of microorganisms are used for carrying out the process of fermentation to produce useful products. They includeBacteria- Acetobacter lacti, Bacillus subtilis, Bacillus polymyxa, Acetobacter woodi etc. Algae- Spirulina maxima, Chlorella sorokiniana, Sorokiniana platensis etc. Fungi- Aspergillus oryzae, Aspergillus niger, Candida utilis etc. Actinomycetes- Nocardia mediterranei, Streptomyces griseus etc. 3. Fermentation Medium: Growth media are required for industrial fermentation, since any microbe requires water, oxygen, an energy source, a carbon source, a nitrogen source and micronutrients for growth. Carbon & energy source + nitrogen source + O2 + other requirements Biomass + Product + byproducts + CO2 + H2O + heat. Nutrient Raw Materials Carbon Source Glucose corn sugar, starch, cellulose Sucrose sugarcane, sugar beet molasses Lactose milk whey Fats vegetable oils Hydrocarbons petroleum fractions Nitrogen Source Protein soybean meal, corn steep liquor, distillers' soluble Ammonia pure ammonia or ammonium salts urea Nitrate nitrate salts Phosphorus source phosphate salts Trace elements: Fe, Zn, Cu, Mn, Mo, Co Antifoaming agents : Esters, fatty acids, silicones, sulphonates, polypropylene Buffers: Calcium carbonate, phosphates Growth factors:

Some microorganisms cannot synthesize the required cell components themselves and need to be supplemented, e.g. with thiamine, biotin, calcium pentothenate Precursors: Directly incorporated into the desired product: Phenyl ethylamine into Benzyl penicillin, Phenyl acetic acid into Penicillin G Inhibitors: To get the specific products: e.g. sodium barbital for rifamycin Inducers: The majority of the enzymes used in industrial fermentation are inducible and are synthesized in response of inducers: e.g. starch for amylases, maltose for pollulanase, pectin for pectinase,olive oil and tween are also used at times. Chelators: Chelators are the chemicals used to avoid the precipitation of metal ions. Chelators like EDTA, citric acid, polyphosphates are used in low concentrations. Process of Fermentation: Process of fermentation varies from product to product. Some of these processes for particular material are given belowIn case of Streptomycin production fermentation process is carried by the following phasePhase-1: In the first 24 hours, growth of the organism starts in the form of mycelium. Streptomyces grisus releases NH3 from soyabean. Glucose is utilized slowly in this period. Production of mycelium is very slight in this phase. The PH is around 6.7 or 6.8-7.5. Phase-ii: Streptomycin production is rapid after 24 hours and continues up to 6 or 7 days. Fairly constant mycelium growth occurs during this period. Glucose is used completely from the medium. The PH of the medium is about 7.6 to 8. Phase-iii: After complete utilization of glucose, the growth ceases. Autolysis takes place (the cells are lysed) and ammonia is released. Hence the PH rises. The mycelium produced is taken for the recovery of antibiotic. In case of Tetracycline production fermentation process is carried by the followingThe commercial method of production of Tetracycline is to subject chlortetracycline to simultaneous dechlorination and hydrozination. Chlortetracycline is dissolved in a solvent like methyl cellulose containing palladium, charcoal and tri ethyl amine as catalyst. Hydrogen is passed at room temperature. Hydrogenation is completed in about 20 minutes. Tri ethyl amine is neutralized with HCl and it is removed by pouring the reaction mixture into water where tetracycline base is crystallized. Tetracycline can also be produced by fermentation process using selected strains of Streptomyces aureofaciens. In this process, inhibitors of chloride utilization such as bromide and organic thiol compound are added to the chlortetracycline producing culture. In another method, chloride free medium is used. The fermentation conditions are similar to chlortetracycline. In case of L-lysine production the process is The medium consist of glycerol, corn steep liquor and [NH4]2HPO4. E.coli is grown on the medium under controlled condition PH, temperature and aeration to produce optimum quantity of DAP. The

incubation is done for about three days. After three days DAP decarboxylase is added to convert DAP to L-lysine which is later extracted. In case of cyanocobalamine productionThe final medium is inoculated with 5 % of inoculums and incubated for stipulated time period to produce the mycelium. Animal or plant extract containing traces of cyanocobalamine is added to culture for best production. Sporulation occurs in 4 to 6 days at 280C. These spores can be stored by freeze drying in sealed vial. In case of Lactic acid production by fermentationLactic acid fermentation is carried out in tanks made up of wood or lined with stainless steel to withstand the corrosive nature of lactic acid. The fermentation is carried out at a PH of 5.5 to 6.5 and temperature of 45-500C for 6 days. The medium is agitated continuously to keep CaCO 3 in suspension to neutralize the lactic acid produced. After 6 days, the broth contains 80-90 percent of lactic acid. The temperature is high to avoid the contamination of butyric acid producing bacteria. Biologicals obtained from fermentation: From fermentation technology we get various types of biologicals or chemical goods. Some of these are shown below in a tabular form-

Products Microorganisms Industrial Chemicals Ethanol ( from glucose) Saccharomyces cerevisiae Ethanol ( from lactose) Kluyveromyces fragilis Citric Acid Aspergillus niger Gluconic Acid Aspergillus niger Acetic Acid Acetobacter spp. Lactic Acid Lactobacillus delbrueckii Amino Acids L-lysine Corynebacterium glutamicium MSG Corynebacterium glutamicium Glutamic Acid Corynebacterium glutamicium Vitamins Riboflavin Ashbya gossypi Vitamin B12 Pseudomonas denitrificans

Ascorbic Acid(L-sorbose) Gluconobecter oxidans Enzymes Amylase Aspergillus oryzae Cellulase Trichoderma reesii Invertase Saccharomyces cerevisiae Lipase Saccharomyces lipolytica Protease Bacillus Polysaccharides Dextran Leuconostoc mesenteroids Xanthan Gum Xanthomones compestris Pharmaceuticals Penicillin Penicillin chrysogenum Cephalosporin Cephalosporium acemonium Amphotericin B Streptomyces nodosus Kanamycin Streptomyces kanamyceticus Neomycin Streptomyces fradiae Streptomycin Streptomyces graseus Gramicidin S Bacillus brevis Polymyxin Bacillus polymxa Chloramphenicol Streptomyces venezuelae Erythromycin Streptomyces erythreus Steriodal Transformations Rizopus nigricans Steriodal Transformations Arthrobacter simplex Viva A (adenine arabinoside) Streptomyces antibiotioticus

Comments: Fermentation; which is an ancient process has been using different fields over the time. Using of fermentation in Biotechnology enlarge the scope of it. Various types of materials are synthesized in biotechnology by using fermentation which is used in different useful tasks. In drug development, the application of fermentation is also undeniable. By using fermentation various life savings drugs are synthesized (i.e. antibiotic, enzymes, amino acids etc.). So, from the above discussion we can easily say that fermentation technology is very much important in drug development.