Expert of yeast origin nutrients for Microorganisms Johnny

Expert of yeast origin nutrients for “Microorganisms” Johnny Yin 18 April, 2014

Contents 1 Introduction about Angel yeast Co. , Ltd. 2 Angel yeast extracts – Good nutrient-sources for microoganisms 3 Application of Angel yeast extract

1. Introduction of Angel Yeast Co. , Ltd. 1. 1 About us > 1. 2 Global net work > 1. 3 Our history > 1. 4 Main business > 1. 5 Angel’s value chain > 1. 6 Employees > 1. 7 Advantages >

1. 1 About us • Angel yeast, founded in 1986, is a listed high-tech yeast company in China. • It is one of the largest global yeast and yeast extract manufacturers with 12 factories in China and Egypt up to now.

1. 2 Global network Factories Yeast extract plants Technical centers Countries 12 4 12 140

1. 3 History Project started 1986 No. 1 in Asia Public company 1989 First facilities 2000 2003 First yeast extract facilities 2008 Today Global supplier

The fastest growing company in global yeast industry • Angel yeast is estimated to grow at a CAGR of 20% 2011 -2012, while the global yeast market’s CAGR is to be 8. 6% (Foodnavigator. com, 2012 -04 -05). CAGR 2011 -2012

1. 4 Main business • • Bakery Yeast extract-savory Yeast extract-fermentation Beverage & ethanol Human health Enzymes Animal nutrition

1. 5 Angel’s Value Chain Lab Yeast extract Inactive yeast Technical support Experiment Microorganism Yeast extract Industrial fermentation Package Trading Product Brewing Food Agriculture Environment Chemical Bakery Yeast extract Enzyme Bio-fertilizer Seasoning Inactive yeast Yeast extract Wine yeast Feed Energy Pharm Alcohol yeast Enzyme Yeast extract Yeast cell wall Feed yeast Enzyme Active yeast Yeast extract Inactive yeast

Our global partners

1. 6 Employees> • 1331 employees, 70% of which are professional people • Over 100 researchers work together with universities. • 7 international experts employed as senior technical consultants. Jan Hille ((Netherlands) 　　 Yao Juan Zhijun Rosenberg (Netherlands) Li Xiao Kuang Jinbao He Chongyue Tilak (U. S. ) (Taiwan, China) Li Pei Li

1. 7 Advantages 1. 7. 1 Strong production output > 1. 7. 2 Professional production process> 1. 7. 3 R&D strength > 1. 7. 4 Quality, safety and consistency > 1. 7. 5 Certification> 1. 7. 6 Environment protection > 1. 7. 7 Global professional service >

1. 7. 1 Strong production output > Total output of Yeast is 120. 000 tons, Yeast Extract is Yichang Plant Chongzuo Plant 46. 000 tons. Yili Plant Binzhou Plant

1. 7. 2 Professional production process

1. 7. 3 R&D strength > • 4 sub-headquarters with technical centers • National yeast technology research and promotion center • Post-doctoral scientific research workstation • R & D team led by Ph. Ds and experts • National recognized laboratory CNAS • Basic research and Application research laboratory • Own the first-class test device and pilot plant.

1. 7. 4 Quality, safety and consistency > • EU standard design production line • Stable and consistent quality applies to all production plants • Distributed control system • Siemens PCS 7 process control system • Whole-course CIP cleaning • SAP-ERP Management flat

1. 7. 5 Certifications > BRC CNAS SEDEX ISO 9001 HALAL ISO 14001 KOSHER(OU) ISO 22000

1. 7. 6 Environment protection > • First class environment protection facilities in each plant • The high-concentration organic waste water was made into bio-organic fertilizer.

1. 7. 7 Global professional service > • We do not only provide the best quality products, we also provide technical solutions for all our serving industries • Set up global baking centers • Over 200 application engineers

2. Angel Yeast Extracts – Good nutrient-sources for microoganisms 2. 1 What’s yeast extract > 2. 2 Why choose yeast extract for fermentation industry > 2. 3 Range of Angel’s typical products >

2. 1 What’s yeast extract? Yeast extract is a kind of : • 100% animal free natural ingredient that is made from yeast. • High performance and stable sources of nutrients for microorganisms • Clean and safe raw materials for industrial fermentation and culture media

Process of yeast extract production

2. 1. 1 Molecular weight distribution

2. 1. 2 Profile of Amino acids

2. 1. 3 Profile of free Amino acids and Total amino acids Free Amino acids / Total Amino acids × 100

2. 1. 4 Trace elements Name of elements Ca Mg Zn Fe Cr Mn K Na As Se Pb Concentration（mg/kg） 246 149 86 33 1. 26 0. 08 9172 2022 0. 03 0. 002 0. 19

2. 1. 5 Vitamins Name of vitamins Concentration（mg/kg） Vitamin B 1 2. 7 Vitamin B 2 56. 9 Niacin 336 Vitamin B 6 17. 6 Pantothenic acid 29 Free D-Biotin 500 （μg/100 g）

2. 1. 6 Nucleic acid and Nucleotides

2. 1. 7 Functionality of yeast extract • Increase biomass • Improve the accumulation of metabolites

2. 1. 8 Stability of yeast extract Two key indicators: Total nitrogen and Amino nitrogen, For example: Stability of Yeast extract FM 902

2. 2 Why we choose yeast extract for fermentation industry? • Clean and Safe - Clean and safe natural material for culture media and industrial fermentation - Animal free, no risks of illness - GMO free, Non allergen - Kosher and Halal certified. • Stable quality - Produced with strict quality control system. - Large industry scale production - No quality change in different seasons • High performance - Advanced production technology Beside yeast extract, Angel also produce yeast peptone and inactive yeast for fermentation industry

2. 3 Range of Angel’s typical products Microfiltered yeast extract Nutrients-enriched yeast extract Yeast peptone FM 888 Powder FM 803 Powder FM 505 Powder FP 101 Powder LP 101 Paste Lab, Biopharmaceuticals Probiotics, Starter culture Probiotics, culture media FM 808 Powder Standard yeast extract FM 902 Powder LM 800 Paste LD 00 Autolyzed & Inactive yeast Economical yeast extract Liquid FM 801 Powder YP 101 Industrial fermentation Powder FM 701 Powder LM 815 Paste Antibiotics, APIs Agriculture and environment

Which are the most popular products in lab research? Products FM 888 FP 101

Angel Microfiltered yeast extract FM 888 • Features and Advantages - High solubility - Easily assimilated by microorganisms and cells - Rich in peptides and growth factors - Low impurities • Application - Lab research Dehydrated culture media, Cell culture, Diagnostic media - High value-added industrial fermentation Insulins, Hormones, Hyaluronic acids

Angel yeast peptone FP 101 • Features and Advantages - Good replacement of Animal origin peptone - Could be combined with yeast extract - Rich in peptides • Application - Lab research Microorganisms culture, small scale fermentation - Industrial fermentation Probiotics, Enzymes, Hormones, Organic acids

Which are the most popular products in industrial fermentation? Products LD 00 LM 800 FM 808 FM 902

Angel standard yeast extract FM 808 • Features and Advantages - Good solubility - High nitrogen level - Rich in peptides - Light color • Application - Lab research Dehydrated culture media - Industrial fermentation Insulins, Hormones, Hyaluronic acids, Probiotics

Angel standard yeast extract FM 902 • Features and Advantages - Total soluble in water - High nitrogen level - Cost effective - High consistency • Application - Industrial fermentation For >60 types of fermentation-based products

Angel standard yeast extract FM 902

Angel standard yeast extract FM 902

Angel standard yeast extract FM 902

Angel standard yeast extract FM 902 Industrial fermentation FM 902 Agriculture Amino acid Enzyme Hormone Organic acid Probiotic Nucleotides / Nucleosides Poly saccharide Biological preservatives Antibiotic Mashroom Rhizobia Biological bait Effective Microorganisms Bio-fertilizer Waste water treatment Environmental microorganisms

Angel standard yeast extract LM 800 and LD 00 • Features and Advantages - Paste and Liquid form of yeast extract FM 902 - Rich in peptides - Cost effective - High consistency • Application - Industrial fermentation For >60 types of fermentation-based products

How to judge the quality? Better yeast extract perform better solubility Better yeast extract perform lighter color Better yeast extract perform higher amino nitrogen Yeast extract should contain low level active microbes Less biomass and productivity means poor quality of yeast extract Baker’s yeast extract is better than Brewer’s yeast extract Baker’s yeast extract must get Kosher/Halal certificates Yeast extract contain rich protein Depends True Good solubility means readily assimilated nutrients, but it depends on specific needs. Some fermentation need light color Higher amino nitrogen product is suitable for seed culture and probiotics, not all the fermentation High level active microbes cause contamination 40% of this situation means unsuitable process, condition or culture media formula Brewer’s yeast extract is not stable which come from spent brewer’s yeast Clean and safe Depends It depends on the customers specific needs

3. Application of Angel Yeast Extract 3. 1 Summary of Angel yeast extract application > 3. 2 Some experiments of Angel yeast extract >

3. 1 Summary of Angel yeast extract application Ø Biomass products industry - Starter cultures and probiotics for human food products - Cultivation of mashroom and macrofungi Recommendation: FM 808, FM 803, FM 505, FP 101 Ø Culture media industry and lab reagent - Bench scale research - Dehydrated culture media, Diagnostics, Cell cuture Recommendation: FM 888, FM 808 Ø Bioactive molecules and chemicals industry - Amino acids and peptides - Organic acids, Vitamins, Nucleotides, Enzymes, Alcohol - Vitamins, Antibiotics, Hormones, Biological preservatives - Other types of large scale fermentation Recommendation: FM 808, FM 902, LM 800, LD 00, YP 101 Ø Agricultural and environmental industry - Sewage treatment, Bio-pesticide, Bio-plastic - Probiotics for agriculture and environment Recommendation: FM 902, FM 801, FM 701, LM 815, LP 101

3. 2 Some experiments of Angel yeast extract 3. 2. 1 Morphology of 4 types of typical microorganisms > 3. 2. 2 Some experiments of Angel yeast extract >

3. 2. 1 Morphology of 4 types of typical microorganisms after using yeast extract and other type of organic nitrogen Streptomyces lincolnensis A Lactobacillus bulgaricus B A Bacillus subtilis A B Acetobacterium B “A”—Another type of organic nitrogen “B”—Angel yeast extract A B

3. 2. 2 Angel standard yeast extract FM 808 for Lactobacillus thermophilus growth Strain: Lactobasillus thermophilus Medium: Yeast Extract FM 808 0. 3%，Glucose 2%, etc. Result: Yeast extract FM 808 could improve the p. H: 6. 5 growth of Lactobacillus thermophilus well Angel yeast extract FM 808 Control

3. 2. 3 Angel standard yeast extract FM 902 for the growth of E. coli and Absidia coerulea The growth curve of E. coli, comparison of FM 902 and Brewer’s yeast extract The conversion rate of HC(from RSA), Absidia coerulea, yeast extract FM 902

3. 2. 4 Angel nutrients enriched yeast extract FM 803 for probiotics Strain: Lactobacillus rhamnosus Medium: MRS Angel yeast extract FM 803 Control Strain: Lactobacillus helveticus Medium: MRS Angel yeast extract FM 803 Control Lactobacillus needs complicated nutritional condition for its growth and acid production. Yeast extract is rich in amino acids, small peptides and growth factors which Lactobacillus needs. Yeast extract FM 803 is a kind of free amino acids enriched product and could improve the growth of Lactobacillus.

3. 2. 5 Angel standard yeast extract FM 902 For L-Tryptophan Fermentation culture medium(%): yeast extract FM 902, glucose 2 %, ammonium sulfate 0. 4%, potassium dihydrogen phosphate 0. 15%, etc. Fig. 1 The effect on L-Tryptophan when different ratio of yeast extracts were used Conclusion: FM 902 is rich in small peptides and free amino acids, and it also contains appropriate polypeptides, vitamins and trace elements. Thus FM 902 is suitable for the fermentation of L-Tryptophan, 2% is a proper level of yeast extract ratio.

Technical support Department of Microbial Nutrition Research • The research and development of new products • The optimization of culture media formula • The suggestion of fermentation process • Development of tailor-made products

Test and scale up platform HPLC GC-MS Pilot Plant Atomic Absorption Spectrometer Powder Instrument Platform for fermentation control GC Drawing Instrument Amino acid analyzer Ultrasonic Processors The membrane separation system Spray drying system

Contact information Any question, please feel free to contact me: Johnny Yin Angel Yeast Co. , Ltd. Address: 168 Cheng Dong Avenue, Yichang, Hubei, China. 443003. Email: yinqiang@angelyeast. com; johnnyyinq@gmail. com

THANK YOU!