Principles of Biotechnology INDUSTRIAL BIOTECHNOLOGY WEEKS 8+9

Transcription

1 Principles of Biotechnology INDUSTRIAL BIOTECHNOLOGY WEEKS 8+9

2 Industrial Microbiology Industrial Microorganisms and Product formation involved: 1- Use microorganisms to produce valuable commercial product 2- Enhancement of metabolic reactions 3- Microbial biotechnology : use gene manipulation to yield new microbial product.

3 Biocatalysis Biocatalysis is the word used to describe the actual reactions carried out by microorganisms in industrial microbiology. Industrial Microbiology originated with alcoholic fermentation processes, such as those of making beers and wine. Microbial processes were developed for the production of Pharmaceuticals (such as antibiotics), food additives (amino acids), enzymes and chemicals such as butanol and citric acid.

4 Industrial Microorganisms and Their Products The major organisms used in industrial microbiology are Fungi (yeast and mold) and certain prokaryotes, in particular genus of Streptomyces. Industrial Microorganisms are metabolic specialists, capable of synthesizing one or more product in high yield. Nature is the ultimate source of all strains of microorganisms used in biocatalytic processes.

5 Properties of Useful Industrial Microorganism 1- The organism must be capable of growth and product formation in large scale culture. 2- It should be easily inoculated into the large fermentors. 3- It must grow rabidly and produce the desired product in a short period of time.

6 4- Must grow in relatively inexpensive liquid culture medium obtainable in bulk quantities. - Many industrial microbiological processes use waste carbon from other industries as major or supplemental ingredients for large-scale culture media. A- Corn steep liquor (a product of corn wet milling industry that is rich in nitrogen and growth factors). B- Whey (a waste liquid of the dairy industry containing lactose and minerals).

7 5- Industrial microorganisms should not be pathogenic, especially to human. 6- Should be amenable to genetic manipulation.

8 Examples of Industrial Products Microbial products of industrial interest include : A- the microbial cells themselves- for example, yeast cultivated for food, baking. B- substances produced by cells, this includes : 1- Enzymes such as Glucose Isomerase, important in the production of high fructose syrup. 2- Pharmaceutically active agents such as antibiotics, steroids and alkaloids. 3- Specialty chemicals and food additives such as aspartame ( a food and drink sweetener), inexpensive chemicals produced in bulk- such as ethanol, citric acid and many others.

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10 Primary and Secondary Metabolites A Primary metabolites is one that is formed during the exponential growth phase of the microorganism. Example Ethanol, is a product of anaerobic metabolism of yeast and certain bacteria. Secondary metabolites is one that is formed near the end of the growth phase, frequently at, near, or in the stationary phase of growth.

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12 Characteristics of Secondary Metabolites 1- Not essential for growth and reproduction. 2- The formation of secondary metabolites is extremely dependant on growth condition, especially on the composition of the media. 3- They produced as group of closely related compounds. For example, a single strain of a species of streptomyces has been found to produce over 30 related but different anthracycline antibiotic.

13 Primary and Secondary Metabolism Pathways Most secondary metabolites are complex organic molecules that require a large number of specific enzymatic reaction for synthesis. For example, it is known that at least 72 separate enzymatic steps are involved in synthesis of the antibiotic tetracycline and 25 steps in the synthesis of erythromycine. Non of these reactions occur in primary metabolism. The metabolic pathways of these secondary metabolites arise out of primary metabolism because their starting materials originate from major biosynthetic pathways.

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15 Major Industrial Products for the Health Industry. Antibiotics: Isolation and Characterization, Antibiotics are chemicals produced by microorganisms to kill or inhibit the growth of other microorganisms. They are typical secondary metabolites. Commercially useful antibiotics are produced primarily by filamentous fungi and by bacteria of the Actinomycete Group table 30.2

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18 Industrial Production of Pencillins and Tetracyclines β-lactum Antibiotics: Penicillin and its Relatives

19 Production of β-lactam Antibiotics Penicillium chrysogenum produces Pinicillin G. Other β-lactum Antibiotics: Cephalosporins, ( broad-spectrum antibiotics) product of the fungus Cephalosporium acremonium.

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22 Vitamines and And Amino acids Vitamins are used as supplement for human food and animal feeds, and production of vitamins is second only to antibiotics in term of total sales of pharmaceutical. Most vitamins are produced commercially by chemical synthesis. Few are too complicated to be synthesized inexpensively but can be made by biocatalysis. Vitamin B12 and Riboflavin are the most important of this class of Vitamins to be produced by Bicatalysis.

23 Vitamin B12 Synthesized in nature by microorganisms. Major deficiency of vitamin B12 leads to a sever condition called pernicious anemia, (in human ) characterized by low production of red blood cells and nervous system disorders.

24 Vitamin B12 benefits

25 Vitamin B12 Source

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27 For industrial production members of the bacterial genera Propionpbacterium and Pseudomonas are the main commercial producers. Cobalt is a metal found in vitamin B12 and yields of the vitamin are greatly increased by addition of small amount of cobalt to the culture media.

28 Riboflavin Riboflavin plays important role in enzymes involved in oxidation-reduction reactions in virtually all organisms. Riboflavin is synthesized by many microorganisms, including bacteria, yeast, fungi. The fungus Ashbya gossypii naturally produces huge amounts of this vitamin (up to 7gm/L).

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30 Amino Acids Amino acids have extensive uses in the food industry, as feed additives, in medicine, and starting materials in the chemical industry. Table 30.3 Glutamic acid is the most important commercial amino acids, used as flavor enhancer. Aspartic acid and phynelalanine, are the ingredient of artificial sweetener aspartame, a non-nutritive sweetener of diet soft drinks and other foods sold as low-calories or sugar-free products.

31 Amino Acid Analysis

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33 Lysine, an essential amino acids for humans and certain farm animals, is commercially produced by the bacterium Brevibacterium flavum for use as a food additives.

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35 Steroids and Biotransformation Process Steroids are important animal hormones that regulate various metabolic processes. Some steroids used as drugs in human medicine, example corticosteroids, reduce inflammation. Estrogens and androgenic steroids, play a role in human fertility and some of them can be used therapeutically in the control of fertility or as stimulants for building muscle mass.

36 Steroids/cont. Most steroids are produced industrially by the process of biotransformation. This involves the growth of organisms in large fermentors, followed by the addition at the appropriate time of sterol precursor. The precursor is obtained from inexpensive starting materials, such as stigmasterol, a by-product of soybean industry.

37 Steroid by Biotransformation Microorganisms in fermentor + stigmasterol Steroids

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39 Enzymes as Industrial Products Exoenzymes : Extracellular enzymes, are capable of digesting insoluble polymers such as cellulose, protein and starch. Various enzymes are used in the food and health industries, primarily as dietary supplements, while many others are produced for laundry applications or in the textile industry.

40 Proteases, Amylases, and High Fructose Syrup Enzymes are produced commercially from both fungi and Bacteria. Proteases are the enzymes produced in the largest amount, used as additives in laundry detergents. Other enzymes are used, such as amylases, lipases, reductases. Such enzymes are produced from alkalophilic bacteria such as Bacillus licheniformis. These enzymes, which have ph optima between 9 and 10, remain active at the alkaline ph of laundry detergent solutions.

41 Extremozymes Enzymes from prokaryotes that live in extreme environments. Extremophiles: organisms that produce extremozymes. Extremozymes: Taq polymerases used in PCR. Proteases, amylases, cellulases, pullulanases and xylanases have been isolated and characterized from various hyperthermophiles. Cold-active enzymes from psychrophiles. Salt-tolerant enzymes from halophiles Low ph (acidophiles) or High ph (alkalophiles)

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44 Vinegar production Vinegar: Ethyl alcohol acetic Acid, by the acetic acid bacteria, Acetobacter. Vinegar can be produced from any substance that contain ethanol. Vinegar is used as favoring ingredient in salads and other foods, and because of its acidity, it is also used in pickling.

45 Acetic acid Bacteria (Acetobacter): They are strictly aerobic bacteria. Acetic acid bacteria are quite acid tolerant and are not killed by acidity that they produce.

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47 Citric acid and other organic compounds Citric acid, is widely used in the food industry as supplement in beverages confections and other foods and in some pharmaceutical applications. Other acids include: 1- Itaconic acid used in the manufacturing of acrylic resins. 2- Gluconic acid; used in the form of Calcium gluconate to treat calcium deficiencies in human and industrially a a washing and water-sotining agents. All these acids produced by fungi. *** Citric acid production: The link to Iron. Citric Acid produced microbiologically by the mold Aspergillus niger

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49 Yeast as Food and Food Supplement

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