Microbial Production of ethanol Bakers Yeast: Baker's Yeast Production - The production of baker's yeast is the largest domestic

use of a microorganism for food purposes. Baker's yeast is a strain of Saccharomyces cerevisiae . The strain of the yeast is carefully selected for its capacity to produce Abundant gas quickly, Its viability during ordinary storage, Its ability to produce desirable flavour . The organisms are mixed with bread dough to bring about vigorous sugar fermentation. The carbon dioxide produced during the fermentation is responsible for leavening or rising of the dough . During manufacturing, the strain is inoculated into a medium which frequently contains molasses and corn steep liquor as sources of carbon, nitrogen, and mineral salts. The reaction of the medium is adjusted to pH 4.4 to 4.6. Ethanol Production By Microbes The most efficient microbes are Zymomonas mobilis (fermenting carbohydrate and producing alcohol twice as rapidly as yeasts) and Thermoanaerobacter elhanolicus , a thermophilic bacterium. Corn sugar and plant starch are used as substrates. A two step fermentation is used for conversion of cellulose to ethanol, (i) conversion of cellulose to sugars, generally by Clostridium spp , followed by (ii) (ii) conversion of these sugars to ethanol by yeasts, Zymomonas or Thermo anaoerobacter spp Citric Acid Fermentations Citric acid, which is a key intermediate of the TCA cycle is produced by fungi, yeast and bacteria as an overflow product due to a faulty operation of the citric acid cycle. Among the organisms used for citric acid production, A. nigerhas been the mold of choice for several decades. A variety of carbohydrate sources such as beet molasses, cane molasses, sucrose, commercial glucose, starchhydrolysates etc., have been used for citric acid production. Among these, sucrose, cane and beet molasses have been found to be the best. For citric acid production the raw material is diluted to 20-25 per cent sugar concentration and mixed with a nitrogen source and other salts. The pH of the medium is maintained around five when molasses is used and at a lower level (pH 3.0) when sucrose is used The fermentations are carried out either under surface, submerged, or solid state conditions. In the surface culture method, shallow aluminium or stainless steel pans are filled with the growth medium, inoculated with the fungal spores and allowed to ferment. In the submerged culture method the mold is cultured in fermentors under vigorous stirring and mixing, while in solid state fermentation, the mold is grown over carrier material such as bagasseetc., which is impregnated with the fermentation medium The production of citric acid by A.niger is largely influenced by the concentration of trace metals such as iron, manganese, copper and zinc in the medium. An appropriate concentration of these elements is essential for good acid production. However, an excess is detrimental. To optimize the level of these trace metals, the raw materials are treated with either ferrocyanide , charcoal, chelating agents or catien exchange resins. Addition of methanol at 3-4 % concentration has been found to enhance the yield of citric acid. This fermentation is an aerobic fermentation and, therefore, adequate aeration is essential for successful citric, acid production Amino Acid Fermentations Microorganisms can synthesize amino acids from inorganic nitrogen compounds. The rate and the amount of synthesis of some amino acids may exceed the cells need for protein synthesis, where upon the amino acids are excreted into the medium.

Some microorganisms are capable of producing sufficient amounts of certain amino acids, to justify their commercial production. The amino acids can be obtained from hydrolysing protein or from chemical synthesis, but in several instances the microbial process is more economical. Secondly, the microbiological method yields the naturally occurring L-amino acids Microbial production of amino acids shows two outstanding features which are not usually encountered in the development of other microbiological processes. One is the importance of auxotrophic micro organisms . These microorganisms were known only as useful tools in the analysis of metabolic pathways and in genetics but, today; they are proving of great value. The second feature is that a knowledge of metabolic control mechanisms can now be used to good purpose in industrial microbiological processes. Penicillin Production The mould from which Fleming isolated penicillin, in was later identified as Penicillium notatum . A variety of moulds belonging to other species and genera were later found to yield greater amounts of the antibiotic and a series of closely related penicillins . The naturally occuring penicillins differ from each other in the side chain (R group). Penicillin was produced by a surface culture method early in World War II. Sub-merged culture methods were are exclusively employed. Penicillin production needs strict asceptic conditions. Contamination by other microorganisms reduces the yield of penicillin. This is caused by the widespread occurrence of penicillinase producing bacteria which inactivate the antibiotic. Secondly, penicillin product ion also needs tremendous amount of air. The medium consists of cornsteep liquor, lactose, glucose, nutrient , salts, phenyl acetic acid or a derivative and calcium carbonate as buffer. The medium is inoculated with a suspension of conidia of Penicillium chrysogenum . The medium is constantly aerated and agitated, and the mould grows throughout as pellets. After about seven days, growth is complete, the pH rises to 8.0 or above, and penicillin production ceases. When the fermentation is complete, the masses of mould growth are separated from the culture medium by centrifugation and filtration. The complex process of extracting the penicillin from the clear fluid then begins. The method involves various extractions with organic solvents and recrystallization . Penicillin is assayed to determine its potency before being bottled and sold. Exopolysaccharide Production ( Xanthan And Dextran ) Xanthan gum is a polysaccharide used as a food additive and rheology modifier . It is produced by fermentation of glucose or sucrose by the Xanthomonas campestris bacterium Type Of Fermentation Used Batch culture for 2-3 days pH Suitable For Fermentation: 7.0 Biosynthesis Synthesis originates from glucose as substrate for synthesis of the sugar nucleotides precursors UDPglucose, UDP-glucuronate , and GDP-mannose that are required for building the pentasaccharide repeat unit. This links the synthesis ofxanthan to the central carbohydrate metabolism. The repeat units are built up at undecaprenylphosphate lipid carriers that are anchored in the cytoplasmic membrane. Specific glycosyltransferases sequentially transfer the sugar moieties of the nucleotide sugar xanthan precursors to the lipid carriers. Acetyl and pyruvyl residues are added as non-carbohydrate decorations. Mature repeat units are polymerized and exported in a way resembling the Wzy -dependent polysaccharide synthesis mechanism of Enterobacteriaceae . Products of the gum gene cluster drive synthesis, polymerization, and e xport of the repeat unit. Applications Of Xanthan

In oil industries for oil recovery at spill site In preparation of toothpaste and water based paints etc Dextran Dextran is a complex, branched glucan (polysaccharide made of many glucose molecules) composed of chains of varying lengths (from 10 to 150 kilodaltons ). The straight chain consists of -1 ,6 glycosidic linkages between glucose molecules, while branches begin from -1,4 linkages (and in some cases, -1,2 and -1,3 linkages as well). (For information on the numbering of carbon atoms in glucose, see the glucose article.) Dextran is synthesized from sucrose by certain lactic-acid bacteria, the best-known being Leuconostoc mesenteroides and Streptococcus mutans . Dextran is also formed by the lactic acid bacterium Lactobacillus brevis Type Of Fermentation Used Batch fermentation Production Concept Produced extracellularly in the medium The enzyme dextransucrase acts on sucrose and brings about polymerization of glucose residues and simultaneously liberates free fructose into the medium Application Dextrans are used as blood plasma expanders, for prevention of thrombosis and in wound dressing Dextrans are useful in the laboratory analytical techniques for purification of biomolecules

Aerobic and anaerobic biological processes for stabilization of solid / liquid wastes; Bioremediatio n.
Microbial Biotechnology Degradation of organic wastes, detoxification of industrial wasters and toxic compounds, degradation of oil spills, etc. are some of the services rendered by microbes. Production of biocontrol agents, biofertilizers, etc, also come under industrial microbiology. Aerobic Digestion of Wastes by Microorganisms Microorganisms digest the waste materials aerobically just as they do it in nature. However
since the quantity of organic load is very high in waste waters, the microbial activities are to enhanced artificially. Aerobic digestion system contains a variety of microorganisms like bacteria, fungi, protozoa, viruses, cyanobacteria and algae. However, the last two are phototrophic and represent only on the surface of the treatment systems. For the same reason, their contribution to waste water treatment is rate limited.

Bacteria Bacteria responsible for the removal of about 85-90% of the BOD remaining in the waste water after primary treatment. They can act in the following ways. Zoogloea ramigera helps in surface attachment and flock formation by secreting a mucus-like polysaccharide. It causes the attachment of various bacterial species to the filter surface and forms a Biofilm. This biofilm contatins bacteria, fungi, algae, insect larve and nematodes.

This animal community degrades carbohydrates, proteins, lipids, etc., into CO2, NO3-, SO42-, and PO43-. Beggiatoa, Thiothrix, etc., proliferate and lead to poor settling characteristics of sewage sludge. Heterotrophic bacteria like Sarcina, Pseudomonas, Escherichia, Stapylococcus, Streptococcus, Salmonella, Shigella, Aerobacter, etc., oxidize the organic molecule. Ammonium released from protein and: to NO3- by nitrifying bacteria Nitrosomonas. and Nitrobacter. The presence ofNO3- even though less toxic than NH4+ is still toxic leading to 'blue babies. Nitrifying bacteria multiply very slowly and so a high population of bacteria is to be maintained. Denitrifying bacteria e.g. Alcaligenes, Achromobacter, Micrococcus, PSuedomonas, etc., ultimately remove NO3- from water by converting it into N2. Denitrification produces various oxides of nitrogen also. The denitrification process is anaerobic and so aerobic digestion should follow anaerobic digestion or it should alternate. Fungi Fungi often in association with algae form a biofilm on the surface of floes. They remove nitrogen, phosphorus and other nutrients. Protozoa Protozoans feed on nutrients and bacteria. Thus smaller molecules are converted to larger
protozoans and so their removal becomes easier.

Anaerobic Digestion of Wastes by Microorganisms Bacteria are the predominant organisms responsible for anaerobic digestion and digest lipids, carbohydrates, proteins, etc. into CH4 and CO2, If the waste water contains sulphate it will be reduced to S2-by Desulfovibrio during oxidation of organic compounds. The sulphate here acts as an electron acceptor. Similarly denitrifying bacteria oxidize organic compounds using nitrate as an electron acceptor converting it into N2. Methanogenic bacteria are special in that they contain several co factors not seen in other bacteria. CO2 in the form of HCO3- is reduced to CH4 in a stepwise manner using three of these cofactors.

The cofactors catalysing the reaction are methanoprotein (MP), methanofuran (MF) and coenzyme M(CoM). The last reaction is catalyzed by factor 430 (F430), the prosthetic group of CoM. The reactions are as follows Treatment of toxic wastes before they reach environment

Many of the above technologies used for decontaminating the contaminated sites can also be used for the treatment of toxic wastes, before they reach the environment. This is easier, since with the knowledge about the toxic wastes, it is easier to control the conditions of treatment. This will also prove to be more cost-effective for the industry. In this method, biodegradable waste from an industrial facility can be treated in line using a bioreactor, which is inoculated with contaminant-degrading bacteria. The treated effluent may then be discharged to an appropriate outlet (e.g. sewer). Another similar example is the use of grease eating bacteria in grease traps at restaurants, rather than using caustic chemical or mechanical force to break up grease clogs Treatment of Solid Wastes - Solid wastes can be sorted out into biodegradable and nonbiodegradable components; the latter may be recycled, while the former may be either (i) incinerated or (ii) used for anaerobic digestion. However, the cost of sorting the waste makes it uneconomical (iii) the entire solid waste is usually dumped into pits the process is called landfill. Oil spills cause severe damage to the ecosystems and pose threats of (1) fire, (2) ground water pollution due to percolation, and (3) air pollution due to evaporation. In addition, oil refineries generate huge quantities of oily sludge, a hydrocarbon waste. The US Environmental Protection Agency and the Exxon Company used microorganism to clean up Alaskan beaches contaminated by the Valdez oil spill. In India, a consortium of bacterial species has been developed to combat oil spills and oily sludge; the inoculant is aptly called 'Oilzapper'. Treatment of Liquid Wastes - Liquid wastes are treated by either aerobic or anaerobic digestion in large centralized water reclamation or sewage treatment units. Generally, particulate matter, e.g., grit and fibers, is present in liquid water, which is first removed by sieving or settling down; this solid component is called sludge.

The main objectives of liquid waste treatment are (i) to reduce the amount of organic material remaining in solution phase and (ii) to inactivate pathogenic organisms that may be present in the waste. But nonbiodegradable compounds would pass through the process without being modified. The solids/sludge recovered from liquid waste may be disposed off as follows: (i) used as fertilizer in agricultural fields (about 30% of the around 10 million tons of sludge produced from water treatment in Western Europe is used in this manner), (ii) disposed off into sea (the preferred mode of disposal in U.K.), (iii) used in landfills along with other solid wastes, (iv) incinerated, and (v) disposed on land following sanitation by pasteurisation, thermophilic digestion or irradiation Agricultural use of sludge is limited by the presence of heavy metals, exotic xenobiotics, and pathogenic micro organisms. Similarly, the environmental soundness of sludge disposal in sea has been questioned by man Bioremediation of Contaminated Soil Soil contamination can be bioremedied by any one or more of the following methods: (i) Conventional land-farming techniques used by many oil companies involve placement of 6 to 12 inches of contaminated soil on top of sand/gravel bd in a treatment cell. Nutrients, water, microbes and other amendments are sprayed over the soil, which is periodically tilled, for the microbes to get oxygen. Leachate collected is sprayed back on the contaminated soil. In situ bioremediation of both soil and ground-water contamination In-situ bioremediation of contaminated soil and ground-water can be undertaken in one of the following three ways using naturally occurring microorganisms: (i) Inoculation with nonindigenous bacteria. In this method, the subsurface environment may be inoculated with bacteria, which are naturally occurring but non-indigenous and are capable of degrading the contaminant. This needs to be accompanied with the supply of balanced nutrient formulation for bacterial growth. (ii) Introduction of H2O2/nutrient formulation for indigenous microbes. The sub-surface environment is sometimes not inoculated with bacteria, since indigenous microbes occurring in sub-surface environment, are often themselves suitable for bioremediation; but the growth of these bacteria is limited due to lack of supply of nutrients and oxygen. Therefore, periodically we may introduce a hydrogen peroxide/nutrient formulation in the sub-surface environment, to help the bacteria grow and metabolize the contaminants to their end products

(iii) Treatment of contaminants in bioreactors. This method involves flushing of contaminants from the soil, so that the same can be treated in bioreactors, where a continuous supply of nutrients, oxygen and microorganisms is maintained. The above three methods can be used individually or in combination, thus providing a range of in sity bioremediation methods Bioremediation of contaminated surface water (pits, ponds and lagoons) only Contaminated surface water can be treated by any one of the following two methods: (i) In the first method, large aerators are placed over the lagoons throughout. It provides sufficient oxygen on the surface to meet the biochemical oxygen demand (BOD) of the organic contaminants for five days. Nutrients and bacterial amendments may be added to these surface waters, whenever necessary (ii) In the second water. The effluent from the bioreactor is either discharged away from the treated water or recycled again (ii) Soil-slurry biotreatment is the second method of choice and involves the use of a vessel containing soil-water slurry (contaminated soil making 1-30% of slurry by volume). Nutrients are regularly supplied and soil-wter slurry is vigorously agitated to allow aeration. (iii) Bioreactor technology, as described in the previous section above, can also be used for bioremediation of contaminated soils Microbial Bioremediation - Bioremediation of organic contaminants is primarily based on either microorganisms naturally present at the sites, or on microbial inoculants developed in the laboratory and introduced at the site. Certain bacterial, fungal and algal species are capable of accumulating some toxic inorganic contaminants as well. However, there is no cost-effective method of removing these microorganisms from the soil after they have sequestered the inorganic ions. Therefore, bioremediation of inorganic contaminants is primarily based on suitable plant species. Biodegradation of Xenobiotics and Toxic Wastes - Bacteria can be modified genetically to develop catabolic pathways for degradation of xenobiotics (wastes from nonbiological systems) and other waste material. Bacterial genes for this purpose arc isolated from bacterial strains (or their plasmids) found at waste sites. Most of these genes are available front gram negative bacteria (e.g. Pseudomonas spp.), which are themselves not very efficient degraders, since multiple genes may sometimes be needed for efficient biodegradation. Therefore, for efficient biodegradation, efficient degraders have to be prepared through genetic engineering and they need to be established in the environment.