1. Yeast extract for Fermentation Xiao Li Ph. D Angel Yeast Co., Ltd 4.19.2014(In Vietnam)

2. Contents Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Advances in modern fermentation regulation technology modern fermentation regulation technology 1 3 2

3. Comparison of typical organic nitrogen source Part 1 Comparison of typical organic nitrogen source. Comparison with different organic nitrogen sources B. Comparison with different organic nitrogen sources Summary of organic nitrogen source A. Summary of organic nitrogen source

4. yeast exact yeast peptone beef extract (powder) corn steep liquor corn steep liquor (powder) cottonseed powder yeast powder soybean powder peanut powder Category others Microbe sources Plant sources Animal sources tryptone

5. Plant sources Other sources Summary of organic nitrogen source A. Summary of organic nitrogen source Animal sources corn steep liquor : by-product of corn starch production ; made from corn soaking water with sulfite after concentration cottonseed meal: oil manufacture by smashed cottonseed cakes beef extract: fresh beef is processd with eliminating fat, hydrolysis, filtration, Concentration Tryptone: casein hydrolyzed by Trypsin. Microbe sources YE ： yeast processed by autolyze, enzymolysis, concentration and drying Yeast powder: inactivated yeast; especially contains cell wall polysaccharides

6. Typical organic nitrogen organic nitrogen yeast exact (Microbe sources) corn steep liquor (Plant sources) beef extract (Animal sources) Advantages efficient;clean Safety; high bioavailability rich in Phosphorus, proline, arginine, phyticacid (ex.:promote the fermentation of penicillin, gentamicin and also can promote the growth of lactobacillus) rich in creatine, creatinine, probably has a postive effect on bacteria growth Disadvantages no reported at present Not stable Contain allergens Not stable Not safe

7. The advantages of yeast extract as nitrogen source YE In favor of cleaner production easy to be utilized safety safety as been widely used as efficient microbial medium in fermentation field comprehensive and balanced nutrients

8. Fermentation Engineering Medicine 、 Food 、 Agriculture 、 Environmental protection 、 Energy field… The important content and key technology in the field of medicine and food Food Additives 、 preservative 、 Functional food Animal cells culture new drugs 、 vaccine 、 enzymes…… Biological energy cellulosic ethanol, biodiesel … Environmental protection: biological treatment 、 biological forecast … Agriculture Deep processing of agricultural products; Biological pesticide and fertilizer... …… Microbial medicine Antibiotics 、 vitamins 、 GM products Biochemicals degradable plastics 、 Bio-PE……. Advances in Modern fermentation regulation technology Part 2 Advances in Modern fermentation regulation technology

9. Metabolic engineering Research directions of Fermentation Optimization of fermentation process Advances in modern fermentation regulation technology Part 2 Advances in modern fermentation regulation technology

10. A. Study on microbial metabolism(multi-scale research) Gene scale Cell scale Engineering scale Scale-up research Scale-up research combined with computer analysis to achieve key regulation parameters

11. Advances in modern fermentation regulation technology Part 2 Advances in modern fermentation regulation technology Optimization of culture medium Optimization of batch fermentation Optimization of fed-batch fermentation Optimization of continous fermentation Study on the theory of fermentation regulation B. Optimization of fermentation process

12. The role of nitrogen sources in fermentation process A. B. Requestments for nitrogen sources in Microbial fermentation C. C. Requestments for YEF in Microbial fermentation Part 3 Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E. E. *Applications in different fermentation fields Reasonable collocation of different nitrogen source in fermentation Reasonable collocation of different nitrogen source in fermentation D.

13. nitrogen source Cell growth organic nitrogen source ： corn steep liquor 、 soybean powder 、 urea 、 *YE 、 … inorganic nitrogen ： Ammonium sulfate 、 Ammonium nitrate 、 Ammonia 、 … carbon source ： CO 2 ↑ A. The role of nitrogen sources in fermentation process Part 3 Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Product metabolite 、 biotransformation 、 enzyme 、 biomass

14. Genetic engineering vaccine products Part 3. Applications of organ i c nitrogen sources for fermentation organ i c nitrogen sources for fermentation B. Requestments for nitrogen sources in Microbial fermentation 1.rich in quick- acting nutrition 2.demand for Special nutritional factors 1.rich in quick- acting nutrition 2.demand for Special nutritional factors Primary metabolic products Secondary metabolic products 1.Reasonable composition of quick and slow-released nitrogen source 2.The limit of special nutritional factors 1.Reasonable composition of quick and slow-released nitrogen source 2.The limit of special nutritional factors 1.Biological safety 2.Fastidious for nutrients 3.Little influence on followed extraction 1.Biological safety 2.Fastidious for nutrients 3.Little influence on followed extraction

15. Requestments for YEF in Primary metabolic products fermentation Requestments for YEF in Primary metabolic products fermentation Requestments for YEF in secondary metabolic products fermentation Requestments for YEF in secondary metabolic products fermentation Requestments for YEF in genetic engineering vaccine products fermentation Requestments for YEF in genetic engineering vaccine products fermentation Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation C. Requestments for YEF in Microbial fermentation

16.  quick-acting nitrogen source ： AN and corn steep liquor could promote the biomass, but inhibit secondary metabolite synthesis.  Slow-release nitrogen source : soya bean powder extend the phase of secondary metabolite synthesis. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation D. Cases: reasonable collocation of different nitrogen source in fermentation

17. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E. Applications in different fermentation fields Biological vaccines Probiotics Vinegar Enzyme preparations Antibiotics Biological mask ( bacterial cellulose By Acetic acid bacteria fermentation ) FermentationFields Amino acid, organic acid, hyaluronic acid, nisin, Biomass energy …….

18. fermentation × 50L tank 5%;0.5-1VVM, 500-700rpm,37 ℃ 20-28h Amplication fermentation × 60m 3 、 × 200m 3 tank…… Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E.1 Bacillus subtilis E.1 High cell density culture process of Bacillus subtilis Activated slant strains Seed culture pH7.0, 37 ℃， 14h 225rpm Shake flask (30/250ml) inoculating 1 loop Seed liquid culture 37 ℃， 220rpm Shake flask 18-20h 200/1000ml inoculums: 5% Optimum medium ， conditions ， fed- batch strategy AchieveHCDC

19. E.1 Bacillus subtilis E.1 High cell density culture of Bacillus subtilis Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Seed medium: Peptone 5 g, yeast extract FM902 6g, NaCl 5g, water 1L.pH7.0—7.2. The optimal fermentation medium Ⅰ (mainly used in research): rice straw powder 200g, wheat bran 800g,supplemented with glucose 50g, Tryptone 20g, yeast extract FM808 20g, NH 4 NO 3 15g, KH 2 PO 4 10g, water, 1500g.. pH7.0—7.2. The optimal fermentation medium Ⅱ (mainly used in industry): glucose 40g/L, yeast peptone FP101 10g/L, yeast extract FM902 35g/L, KH 2 PO 4 15g/L, CaCO3 25 g/L. pH7.0—7.2.

20. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Drying process Fermented broth centrifuged paste Powder mixed with CaCO 3 as feed Spray drying Fermented broth+dextrin Spray drying Powder as feed Fermented broth centrifuged paste + Bran+ CaCO 3 Powder stoving

21. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E.2. fermentation E.2. Vinegar fermentation (1)Raw material: rice vinegar, bran vinegar, fruit vinegar, wine vinegar, substitute material vinegar etc. (2)vinegar fermentation process: Solid-state ; liquid fermentation ;solid-liquid fermentation (3)color and luster of the product: Smoked vinegar; Pale vinegar; White vinegar Category

22. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E.2. fermentation E.2. Vinegar fermentation

23. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation (1) Traditional fermentation (1) Traditional Vinegar fermentation

24. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation (2) Improved fermentation (2) Improved Vinegar fermentation Nutritive salts nutrients

25. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation (3) Alcohol fermentation (3) Alcohol Vinegar fermentation Packing Cooling water tower Cooling water inlet Nutritive salts edible alcohol water charging stock tank Fermentor Product mix Filtration discharge tank Warehouse Cooling water outlet YE Water

26. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Theory of producing Theory of producing Vinegar

27. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Theory of producing Theory of producing Vinegar

28. Nutritive salts in Vinegar fermentation Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E.2. fermentation E.2. Vinegar fermentation The vinegar Nutritive salts contains essential salts and growth factor which could promote growth and metabolism for acetic acid bacteria and effectively shorten the fermentation period, increase production, improve the quality of acetic acid and reduce the cost of the production.

29. Acitic acid fermentation ADY α-Amylase Glucoamylase Nutritive saltsYE Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation Provide product scheme to you

30. References Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation E.2. fermentation E.2. Vinegar fermentation 1.Research on Optimization of Fermentation Medium for Alcohol Vinegar Production by Uniform Design ( Key Laboratory of Industrial Biotechnology of Ministry of Education jiangnan University ， Wuxi 214122 ， China) ABSTRACT :The experimental data were analyzed by stepwise regression to obtain a linear regression model for optimized fermentation media: Glucose 5 g /L ， yeast powder 3 g /L ， NaH 2 PO 4 0. 7 g /L ， KH 2 PO 4 0. 6 g /L ， MgSO 4 0. 4 g /L ． Under these conditions ， a batch fermentation conducted in 5 L tank for 25 h ， which obtained 0.42 g /L of biomass ， 5.76 g /100 mL acetic acid ． After that ， semi-continuous fermentation were acted ， 95% conversion rate of ethanol ． The productivity was 0. 2 g /( 100 mL·h) ， which had achieved the international level of production of alcohol vinegar ．

31. E.3. Lincomycin fermentation  Lincomycin, a member of the lincosamide group of antibiotics, is produced by Streptomyces lincolnensis, and it is used for the treatment of diseases caused by Gram-positive bacteria. At present, Lincomycin is produced by many factories at home and abroad, but the strain is easy to deteriorate, which would lead to the decline of production. Furhtermore, the biosynthesis route and regulation mechanism of lincomycin are still unclear, so it would be a complicated task to optimize and scale-up the fermentation process. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation

32. E.3. Lincomycin fermentation Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation

33. E2. Lincomycin fermentation  Seed medium （ % ）：  Starch 1.4 ， Glucose 2.8 ， Soya bean powder 2.0 ， FM802 2.6 ， NaCl 0.07 ， NaNO3 0.085 ， CaCO3 0.7 ， NH4NO 3 0.2 ， (NH4)2SO4 0.23 ， KH2PO4 0.0045%, initial pH 7.3.  Fermentation medium （ % ）  Starch 1.0 ， Glucose 4.0 ， Soya bean powder 2.78 ， FM902 0.3 ， Yeast power YP101 0.6, NaCl 0.52 ， NaNO3 0.62 ， KH2PO4 0.026 ， CaCO3 0.74 ， NH4NO3 0.21 ， (NH4)2SO4 0.21, initial pH 7.8. Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation

34. E2. Lincomycin fermentation Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation

35. E2. Lincomycin fermentation Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation

36.  in the early stage Part 3. Applications of organic nitrogen sources for fermentation organic nitrogen sources for fermentation vacuoles Hyphae were broken

37. Yeast extract FM902 was added to the broth, andμwas kept at about 0.02 In the mid-late and late stage

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