DR . Hanan M. A. Abdel Razek Biotechnology

The objectives of BIOTECHNOLOGY are: Establishment of infrastructure and facilities for Life Science and Biotechnology R&D in universities or institutions Encouraging the utilisation of the established infrastructure on shared or service basis Establishment of National Repositories: Repositories for biomaterials ensure A continuous supply of high-quality viruses, bacteria, and fungi, as well as cell lines Genetic stocks and advanced sequencing facilities Promotion of development of indigenous biochemical reagents Clinical and field trial facilities Access to synchrotron beamlines Animal house facilities National Primate Research Centres Bioclusters GLP certified laboratories Viral and regulatory testing facilities Laboratories for biosafety assessment Bio-incubators Biotech parks National Gene Vector Laboratories Islet Cell Resource Centres Human tissue and organ resource and animal biotechnology platforms Noninvasive imaging and spectroscopy.

Teach, train and graduate qualified skilled scientists in many disciplines of Biotechnology. Develop, implement and evaluate the curriculum of biotechnology program with a strong commitment to be aligned and comply with the national and international standards of the quality assurance. Conduct applied research in basic and translational biotechnology and fosters interactions and collaboration between faculty, researchers and industry. Promote a supportive learning environment for life through implementing continuous learning programs. Apply the general principles and standards of technology transfer and ensure adequate training of our personnel in modern biotechnology.

Microbes are usually preferred than plant and animal cells •- growing on a wide range of substrate •- useful metabolic pathway •- easily manipulated for desired metabolic pathway •-easily manipulated genetically •- higher growth rate compared to other organisms Generation time ( time required for cell division) of E.Coli → 15 – 20 min Plant cell → 12 hrs Animal cell → 2-3 days ( about 60 hrs) •-M.O are available in all time and has suitable space

scope or resources of biotechnology A- Microbial Biomass •(1) SCP ( single cell protein ) • sources: production of SCP from waste materials ? • it removes pollution and generate good quality of SCP •Substrate: alcohol, molasses and whey e.g. use of yeast cell in baking •(2) Mushrooms: fruiting bodies of filamentous fungi •Advantages: conversion of plant wastes into edible food –Possess good flavor –Separation of biomass is easy

B- Production of microbial polymers •Dextran is polymer of CHO produced by ---- Leuconostoc mesenteroids ----used as plasma substitute •Pullulan ---- Aureobasidium pullulans ---- used as biodegradable film C-Production of vitamins and amino acids, antibiotics and organic acids •Riboflavin ( vit B 6 ) ---- produced by Ashbya gossypi . ---- used in vitamin deficiency as in diabetic pateint •Cyanocobalamin ( vit B 12 ) ---- produced by Propionobacterium sp. ---- used in Pernicious anemia •Glutamine ( a . a ) ---- produced by Corynebacterium sp. ---- used as Food supplement •Penicillin ( antibiotic) ---- produced by Penicillium chrysogenum ---- used as antibiotic •Citric acid ( organic a ) ---- produced by Aspergillus niger ---- used as effervescent •D- Iron chelating agent •Desferrioxamine B ---- produced by Streptomyces ---- antidote for the treatment of acute iron poisoning

E- Microbial enzymes •Industrial enzymes ( enzymes produced by M.O and used in industry ) ---- pectinase, cellulase and amylase ---- used in food industry •Restriction endonuclease ---- used in recombinant DNA technology for synthesis of insulin , interferons, erythropiotein, growth hormones and DNA polymerase •lipase and protease ---- used in Detergent industry Application of commercial microbial enzymes (ME) •Detergent , starch production, cheese, plant juice •Analytical, diagnostic and therapeutic use of ME e.g ↓ •ME , MO use •Amylase(analytical, therapeutic ) ----Aspergillus, Bacillus ---- digestive aids •L-asparaginase (therapeutic ) ---- E. coli ---- cancer chemotherapy •Catalase (therapeutic ) ---- Aspergillus niger ---- contact lens cleaning •Glucose oxidase (diagnostic )---- Aspergillus niger ---- Blood glucose analys •Streptokinase (therapeutic ) ---- streptococci ---- liquifying blood clot •Urease(analytical ) ---- lactobacillus ---- urea removal

•3- product is free from enzyme Immobilizing enzymes How can increase the capacity of enzymes ---- By immobilization ---- means carried enzyme on carrier to work for long time instead of short time 1- reused 2-suitable for continuous application •3- product is free from enzyme 4-improved stability mean this enzyme not affected by change in pH, temp…..etc

•1- Production of antibiotics F. Application of m.o in partial synthesis of pharmaceutical Products •1- Production of antibiotics •Hydrolysis of benzylpenicillin by ------- penicillin acylase and ------- Production of 6-amino-penicillanic acid ( synthetic penicillin) •Production of 7-amino-cephalosporanic acid •2- Steroid transformation •Progestrone ------- 11-alpha hydroxyprogestrone By Rhizopus nigricans (fungi) •Common transformation reaction •1- hydroxylation 2-dehydrogenation 3- hydrogenation 4- epoxidation 5- side chain cleavage 6- degradation •Examples •Hydrocortisone ---- prednislone •Progestrone------- 11-alpha hydroxyprogestrone

•MICROBIAL FERMENTATION •Fermentation Industry •MICROBIAL FERMENTATION •1- Production of microbial biomass •2- Production of microbial enzymes •3- Production of microbial metabolites •4- Transformation reactions •Bacterial fermentation •1- lactic acid Lactobacillus ( Lactic acid is commonly used in food industry) •2- Vit B12 Propionobacterium sp.

kinds of fermenters • •1- Stirred tank fermenter ------ > antibiotic production •2- Tubulat tower fermenter---- > production of wine, vinegar, beer etc. •3- Internal recycle air-left fermenter -- > yeast production from oil •4- External recycle air-left fermenter -- > production for bacterial biomass from methanol •5- Nathan fermenter -- ------ --------> brewing industry

Fermentation technology Fermentation (old) ……. Glucose molecules act as electron donner and electron acceptor Fermentation (new)….. Growing of MO on large scale e.g penicillin fermentation Control of O2 level in the cultural growth of Saccharomyces (e.g hexose) yield different product: aerobic Hexose ---------------- backer’s yeast anaerobic Hexose ------------------------ alcohol

Characters of organisms to be used in biotechnology 1- be grown in pure culture, 2- genetically stable, -amenable to genetic manipulation, -grow rapidly on non-expensive raw materials, -Limited or no need for vitamins and other growth factors ( nitrogenous compound e.g vitamins , a a , which the microbial growth mainly depend on the presences of these compound) 6-efficient production of the target product, -safe and non-pathogenic, -ready harvesting from the fermentation medium, 9- grow well under workable conditions (pH, Temp, et.) 10- production of limited by-product( not need step of purification of product)

Improvement of product quality and quantity by manipulation of microorganism upstream manipulations such as: a. Screening program to select the active strain ( i.e to select optimum productivity) b. Improvement of selected strain by mutation through change of genetic information (maximum yield by maximum gene expression through transfusion and trans duction is the process by which foreign DNA is introduced into a cell by a virus or viral vector. An example is the viral transfer of DNA from one bacterium to another.) c. Improvement of the existing process and products by recombinant DNA technology (Insulin production by culture on E coli to produce insulin) d. The use of cell fusion and cell hybridization for generation of cell hybrides with high productivity (hybridization method Nitrogen fixation and monoclonal antibody) in hybridization the 2 cell become one cell to take best character of the 2 cell leading to increased productivity

1- needs less space and labor Maximization of the yield through manipulation of the fermentation conditions and down stream processing, which include: 1. Manipulation of the culture conditions to specify product and maximize yield a. Control of O level in the cultural growth of Saccharomyces yield different product: aerobic Hexose ---------------- backer’s yeast) biomass) anaerobic Hexose ------------------------ alcohol b. Citric acid prod. by Aspergillus niger spores could be performed by surface or submerged culture. Submerged culture is preferred due to: ( is suitable for fungi more than bacteria) 1- needs less space and labor 2- more economic with high yield due to high contact between media 3- ease of sterilization, where low energy is required c. Removal of metal cations (Fe+2 & Mg+2) from the media for citric acid production usually improve the yield ) metals must be removed because its toxic to MO and chelating citric acid leading to decreased production ( . 2- Instruction of mathematical modeling for the fermentation parameters, helping in prediction of the best conditions for maximizing yield, cost reduction and scaling up the process ) requirement lab study flask, pilots study yield 3-5, field study in fermenter yield more than 100 product

3- Improvement of the capacity of the biocatalyst (microorganisms 3- Improvement of the capacity of the biocatalyst (microorganisms. or enzyme) e.g. by immobilization for maximizing the yield and the production Immobilization means increased capacity of microorganisms. or enzyme

4- Using proper methods for separation and isolation, recovery and purification of the product to maximize the yield and reduce lost during extraction and purification (ion exchange and chromatography)----- downstream processing Industrial fermentations comprise upstream processing (USP) and downstream processing (DSP) stages USP involves all factors and processes leading to and including, fermentation (i) Aspects related to the producer organism selecting the suitable producer industrial strain improvement to enhance productivity maintenance of strain purity preparation of suitable inoculum development of selected strains to increase economic efficiency Fermentation media - (ii) (iii) Fermentation conditions developed to optimize growth or the desired product

downstream processing manipulation by upstream processing manipulation by Using proper methods for separation and isolation, recovery and purification of the product to maximize the yield and reduce lost during extraction and purification (ion exchange and chromatography) Centrifugation crystalization Improvement of Microbial culture Fermentation media Fermentation condition

Downstream processing includes all processing following fermentations - aim of : 1- efficiently, reproducibly and safely recovering the target product to the required specifications (activity, purity, etc.), 2- while maximizing recovery yield and minimizing cost. (i) primary recovery (ii) product purification (iii) finishing processes )crystallization) • Role of genetic engineering in DSP DSP can be improved by : i- modification of the organism to suppress by-product formation, ii- recombinant protein products may be designed to be excreted outside the cell , ) product outside the cell preferred to prevent cell lysis) microbial cell wall may be modified to increase its permeability to the product iv- recombinant protein products may be engineered with a high affinity for certain separation matrix. iii-

Role of genetic engineering in USP 1- genetic fusion, 2- protoplast fusion, 3- gene amplification 4- recombinant DNA technology

Questions

Thank You DR . Hanan M. A. Abdel razek Please study well : - )