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

Yeast extract for Yeast extract for FermentationFermentation

Xiao Li Ph. D Angel Yeast Co.,

Ltd4.19.2014(In

Vietnam)

Contents

Applications Applications ofof organic nitrogen sources for fermentationorganic nitrogen sources for fermentation

AdvancesAdvances in in modern fermentation regulation technologymodern fermentation regulation technology

1

3

2

Part 1 ComparisonComparison of of typical organic nitrogen source typical organic nitrogen source

B . . Comparison with different organic nitrogen Comparison with different organic nitrogen sourcessources

A . Summary of organic nitrogen sourceSummary of organic nitrogen source

A . Summary of Summary of organic nitrogen source organic nitrogen source

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 source

sAnimal sources

tryptone

Plant sources

Other sourcesOther sources

A . Summary of Summary of organic nitrogen organic nitrogen sourcesource

Animal sources

corn steep liquor :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,ConcentrationTryptone: casein hydrolyzed by Trypsin.

Microbe sources

YE ： yeast processed by autolyze, enzymolysis, concentration and drying

Yeast powder: inactivated yeast; especially contains cell wall polysaccharides

Typical

organic organic nitrogen 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

The advantages of yeast extract as nitrogen sourceThe advantages of yeast extract as nitrogen sourceThe advantages of yeast extract as nitrogen sourceThe advantages of yeast extract as nitrogen source

YE

In favor of In favor of cleaner cleaner

productionproduction

easy to easy to be be

utilizedutilized safetysafety

as been widely used as efficient microbial medium in fermentation field

comprehensive comprehensive and balanced and balanced

nutrientsnutrients

Fermentation EngineeringFermentation Engineering

Medicine 、 Food 、 Agriculture 、 Environmental protection 、 Energy field…Medicine 、 Food 、 Agriculture 、 Environmental protection 、 Energy field…

The important content and key technology in the field of medicine and food

FoodAdditives 、 preservative 、 Functional food

Animal cells culturenew drugs 、 vaccine 、enzymes……

Biological energy

cellulosic ethanol , biodiesel ……Environmental

protection:biological treatment 、 biological

forecast…

AgricultureDeep processing of agricultural products;Biological pesticide and fertilizer...

…………

Microbial medicineAntibiotics 、 vitamins 、GM products

Biochemicalsdegradable plastics 、 Bio-PE…….

Part 2 AdvancesAdvances in Modern fermentation in Modern fermentation

regulation technologyregulation technology

Metabolic Metabolic engineeringengineering

Research directions of Research directions of FermentationFermentation

Optimization Optimization of of

fermentation fermentation processprocess

Part 2 AdvancesAdvances in modern fermentation in modern fermentation




A . Study on microbial metabolism(multi-scale research)A . Study on microbial metabolism(multi-scale research)

Gene scale

Cell scale

Engineering scale

Scale-up researchScale-up research

combined with computer analysis to achieve key regulation parameters



Optimization of culture medium

Optimization of batch fermentation

Optimization of fed-batch

fermentation

Optimization of continous

fermentation

Study on the theory of Study on the theory of fermentation regulationfermentation regulation

B. Optimization of fermentation process

The role of nitrogen sources in fermentation processA.

B. Requestments for nitrogen sources in Microbial fermentation

C.C. Requestments for YEF in Microbial fermentation

Part 3 Part 3 Applications Applications ofof organic nitrogen sources for fermentationorganic nitrogen sources for fermentation

E.E. *Applications in different fermentation fields*Applications in different fermentation fields

RReasonable collocation of different nitrogen source easonable collocation of different nitrogen source in fermentationin fermentationD.D.

nitrogen source

Cell growthCell growth

organic nitrogen source ：corn steep liquor 、 soybean powder 、 urea 、 *YE 、…

inorganic nitrogen ：Ammonium sulfate 、 Ammonium nitrate 、 Ammonia 、…

carbon source ： CO2↑

A. The role of nitrogen sources in fermentation process

Part 3 Part 3 Applications Applications ofof organic nitrogen sources for fermentationorganic nitrogen sources for fermentation

Productmetabolite 、 biotransformation 、

enzyme 、 biomass

Genetic engineering

vaccine products

Part 3. Part 3. Applications Applications ofof

organorganiic nitrogen sources for fermentationc 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

Requestments for YEFin Primary metabolic products fermentation

Requestments for YEFin Primary metabolic products fermentation

Requestments for YEFin secondary metabolic products fermentation

Requestments for YEFin secondary metabolic products fermentation

Requestments for YEFin genetic engineering vaccine products fermentation

Requestments for YEFin genetic engineering vaccine products fermentation


organic nitrogen sources for fermentationorganic nitrogen sources for fermentation

C. Requestments for YEF in Microbial fermentation

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.



D. Cases:D. Cases: reasonable collocation of different nitrogen source reasonable collocation of different nitrogen source in fermentationin fermentationD. Cases:D. Cases: reasonable collocation of different nitrogen source reasonable collocation of different nitrogen source in fermentationin fermentation



E. E. Applications in different fermentation fieldsApplications in different fermentation fieldsE. E. Applications in different fermentation fieldsApplications in different fermentation fields

Biological vaccines

ProbioticsVinegar

Enzyme preparationsAntibiotics

Biological mask(bacterial cellulose

By Acetic acid bacteria

fermentation)

Fermentation Fermentation FieldsFields

Amino acid, organic acid, hyaluronic acid, nisin, Biomass energy …….

fermentation× 50L tank

5%;0.5-1VVM,500-700rpm,37℃

20-28h

Amplication fermentation× 60m3 、

× 200m3tank……



E.1 E.1 High cell density culture process of Bacillus subtilis Bacillus subtilis E.1 E.1 High cell density culture process of Bacillus subtilis Bacillus subtilis

Activated slant strains

Seed culturepH7.0,

37℃ ， 14h225rpm

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

Achieve Achieve HCDC HCDC

E.1 E.1 High cell density culture of Bacillus subtilis Bacillus subtilis E.1 E.1 High cell density culture of Bacillus subtilis Bacillus subtilis



Seed medium: Peptone 5 g, yeast extract FM902 6g, NaCl 5g, water 1L.pH7.0—7.2.

The optimal fermentation medium (mainly used in research)Ⅰ : rice straw powder 200g, wheat bran 800g,supplemented with glucose 50g, Tryptone 20g, yeast extract FM808 20g, NH4NO315g, KH2PO410g, water, 1500g.. pH7.0—7.2.

The optimal fermentation mediumⅡ (mainly used in industry): glucose 40g/L, yeast peptone FP101 10g/L, yeast extract FM902 35g/L, KH2PO415g/L, CaCO3 25 g/L. pH7.0—7.2.



Drying process

Fermented broth

centrifuged

paste Powder mixed with

CaCO3 as feed

Spray drying

Fermented broth+dextrin

Spray drying Powder as feed

Fermented broth

centrifugedpaste +Bran+CaCO3 Powder

stoving



E.2. E.2. Vinegar fermentationfermentationE.2. E.2. Vinegar fermentationfermentation

(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

CategoryCategory






(1) Traditional (1) Traditional Vinegar fermentationfermentation(1) Traditional (1) Traditional Vinegar fermentationfermentation



(2) Improved (2) Improved Vinegar fermentationfermentation(2) Improved (2) Improved Vinegar fermentationfermentation

Nutritive saltsNutritive saltsnutrientsnutrients



(3) Alcohol (3) Alcohol Vinegar fermentationfermentation(3) Alcohol (3) Alcohol Vinegar fermentationfermentation

Packing Packing

Cooling water tower Cooling water tower Cooling water inletCooling water inlet

Nutritive saltsedible alcohol

water

Nutritive saltsedible alcohol

water

charging stock tank charging stock tank FermentorFermentor Product mixProduct mix

FiltrationFiltration

discharge tank discharge tank

WarehouseWarehouse

Cooling water outletCooling water outlet YEWater

YEWater



Theory of producing Theory of producing Vinegar Theory of producing Theory of producing Vinegar



Theory of producing Theory of producing Vinegar Theory of producing Theory of producing Vinegar

Nutritive salts

in 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.

Acitic acid fermentation

ADYα-Amylase

GlucoamylaseNutritive salts YE



Provide product scheme to youProvide product scheme to you

ReferencesReferences




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 ， NaH2PO4 0. 7 g /L ， KH2PO4 0. 6 g /L ， MgSO4 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 ．

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.



E.3. Lincomycin fermentation



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 ， NH4NO3 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.









in the early stage



vacuoles vacuoles

Hyphae were brokenHyphae were broken

Yeast extract FM902 was added to the broth, andμwas kept at about 0.02

In the mid-late and late stage In the mid-late and late stage

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