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BTY323 Lectures 15, 16 Enzymes in Industry

Industrial Enzyme Classes. Commodity enzymesHigh volume (tonnes p.a)Low purity (but not necessarily so)Low cost (e.g. $5-40 per kg)Low profit marginsSpeciality enzymesLow volume (g

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BTY323 Lectures 15, 16 Enzymes in Industry

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    1. BTY323 Lectures 15, 16 Enzymes in Industry Markets Types Scale Values Future Examples

    2. Industrial Enzyme Classes Commodity enzymes High volume (tonnes p.a) Low purity (but not necessarily so) Low cost (e.g. $5-40 per kg) Low profit margins Speciality enzymes Low volume (g kg) High purity High cost ($5 10,000 per g) High profit margins

    3. Enzymes in Industry Distribution of enzymes by substrate Protein hydrolysing 59% Carbohydrate hydrolysing 28% Lipid hydrolysing 3% Speciality (analytical, pharma, research) 10%

    4. Enzymes in Industry

    5. Industrial enzymes: Market trends Increasing 10-15% annually by volume Increasing 4-5% annually by value Decreased margins for commodity enzymes Increased use of speciality enzymes Diagnostic enzymes Fine chemicals manufacture Chiral separation

    6. Industrial enzymes Food processing Textiles Grain processing Amylases in bread-making Lipases in flavour development Proteases in cheese making Pectinases in clarifying fruit juices Cellulases in treating denim to generate stone-washed texture/appearance Conversion of corn starch to high fructose syrups

    7. Industrial enzymes Feed enzymes Waste management Diagnostic enzymes Enzymes to assist in the digestibility of animal feeds (cellulase, xylanase, phytase) Lipases as drain-cleaning agents Reporter enzymes (alkaline phosphatase, glucose oxidase, b-glucosidase) and diagnostic enzymes (DNA polymerase)

    8. Industrial enzymes Speciality Biotransformations Lipases, esterases and oxidoreductases for chiral separations Glucotransferases in synthesis of oligosaccharides Thermolysin in aspartame synthesis Nitrile hydratases in acrylamide and nicotinamide synthesis Proteases in peptide synthesis Penicillin acylase for manufacture of semisynthetic penicillins Aspartase in the manufacture of L-aspartate

    9. Examples of Industrial Enzyme Processes Starch conversions and the production of High Fructose Syrups Aspartame biosynthesis Nitrile conversions Acrylamide Nicotinamide

    10. Corn starch processing 1

    11. Corn starch processing 2.

    12. Enzyme step 1: Action of Termamyl on starch granules Termamyl is an a-amylase (cleaves a-1-4 glucosidic bonds in starch) High temperature expands starch granules, making amylose and amylopectin chains more accessible Termamyl is sufficiently stable at high temperatures if short reaction times are used Starch hydrolysis is a batch process (the enzyme is not reused!)

    13. Enzyme step 2: Conversion of maltose to glucose Amyloglucosidase is not as thermostable as Termamyl (temperature must be reduced) Amyloglucosidase has a pH optimum of 6.5 (Termamyl operates optimally at 8.5): pH must be reduced Reaction kinetics are slower Long incubations result in caramelisation of the saccharides - resulting in product loss and increase in impurities

    14. Enzyme step 3: Conversion of glucose to fructose Fructose is much sweeter than glucose; it can be used as a sweetening agent in foodstuffs, and is more profitable than glucose The enzyme xylose isomerase will convert glucose to fructose, in an equilibrium reaction Glucose ? Fructose A 50:50 mixture of glucose:fructose is sold as high fructose syrup (HFS) Xylose (glucose) isomerase is much less thermostable, and inhibited by Ca ions.

    15. Aspartame biosynthesis

    16. Key process characteristics Immobilised enzyme allows continuous process and enzyme reuse Proteases normally hydrolyse peptide bonds: a low water activity solvent system (organic solvent based) is necessary to reverse the normal equilibrium Organic solvents often promote enzyme denaturation: Thermolysin as a stable thermophilic protease Product recovery is easy the CBZ-L-Phe-L-Asp-OMe intermediate crystallizes out in the reaction media

    17. Column-based biosynthesis of Aspartame Thermolysin is used in a column format Reaction will be run continuously until substrate breakthrough is observed This indicates that the enzyme efficiency is dropping (inhibition or denaturation) Several columns may be operated in series to achieve maximum conversion efficiencies

    18. Nitrile biotransformations

    19. Production of acrylamide

    20. Production of nicotinamide

    21. Other large-scale industrial enzyme processes Penicillin acylase Penicillin (produced at very high yields by industrial-strain Streptomyces fermentations) is converted enzymatically to 6-aminopenicillanic acid 6-Aminopenicillanic acid is a substrate for chemical or microbial conversion to valuable commercial antibiotics (e.g. Ampicillin)

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