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Fermentation Engineering 赵 蕾 Zhaolei, PhD College of Life Science Shandong Normal University E-mail: [email protected] 1 How to study effectively? Give You a “PPT” P: Preview P: Practice T: Taking notes 2 What can we learn from this course? • Knowledge • Approach • Attitude • Team work • Skills 3 Introduction Concept of fermentation History of fermentation Types of fermentation products Steps in industrial fermentation 4 What is fermentation? The term fermentation is derived from the Latin verb fervere, to boil, which describes the appearance of the action of yeast on extracts of fruit or malted grain during the production of alcoholic beverages. 5 Biochemist Fermentation is an anaerobic process where energy is produced without the participation of oxygen or other inorganic electron acceptors. (“life without air”) 6 Microbiologist Any process for the production of a product by the mass culture of microorganisms. 7 Definition of Fermentation Fermentation is the chemical transformation of organic substances into simpler compounds by the action of enzymes, which are produced by microorganisms such as molds, yeasts, or bacteria. Fermentation is a Unique process with great potential Environment friendly Consume less energy Easy to manage 9 History of fermentation Ethanol fermentation was one of the first chemical reactions observed by humans. In nature, various types of food "go bad" as a result of bacterial action. Early in history, humans discovered that this kind of change could result in the formation of products that were enjoyable to consume. 10 Pioneers in Microbiology Anthony van Leeuwenhoek Made the earliest microscopes which magnified up to 300X. Discovered microorganisms. (1632-1723) 11 Leenvenhoek ′ s microscopes Louis Pasteur Showed microbes caused fermentation & spoilage Disproved spontaneous generation of microorganisms Developed aseptic techniques (1822-1895) 14 Robert Koch Developed pure culture methods. Established a sequence of experimental steps to show that a specific m.o. causes a particular disease ——Koch’s Postulates (1843-1910) 15 Single strain 16 The Nobel Prize in Chemistry 1907 "for his biochemical researches and his discovery of cell-free fermentation" Eduard Buchner Germany 1860-1917 17 In 1897, Buchner ground up a group of cells with sand until they were totally destroyed. Then he extracted the liquid and added it to a sugar solution. His assumption was that fermentation could no longer occur since the cells were dead. But he was amazed to discover that the cell-free liquid did indeed cause fermentation. 18 History of Fermentation engineering Start from an ancient time Large scale production Early 20th century – beverage industry, vinegar, baking yeast, citric and lactic acid Late 20th century – development of biotechnology from the antibiotic industrial 19 20 Major Products of Industrial Microbiology Industrial and agricultural products Fermented Food and Food additives Products for human and animal health 21 The product can either be The cell itself: referred to as biomass production. A microorganism own metabolite: referred to as a product from a natural or genetically improved strain. 22 Products from Microorganisms vinegar 23 Q: Why antibiotics , amino acid and enzymes could be produced by microbial fermentation ? 24 Micro-organisms cause changes in the foods which: Extend the shelf life ( preservation ) Producing desirable taste and flavour Improvement of nutritional value Producing required physicochemical properties 25 Types of microbial products Primary metabolites —— produce during growth stage – related to synthesis of microbial cells in the growth phase – include amino acids, nucleotides, fermentation end products, and enzymes 26 26 Secondary metabolites ——produce after growth stage – accumulate following active growth – have no direct relationship to synthesis of cell material and natural growth – include antibiotics and mycotoxins 27 28 29 Primary metabolite Microorganism Significance Ethanol Saccharomyces cerevisiae Aspergillus niger alcoholic beverages Acetone and butanol Clostridium acetobutyricum solvents Lysine and Glutamic acid Corynebacterium glutamacium nutritional additive and flavour enhancer Xanthan gum Xanthomonas campestris industrial Secondary metabolite Microorganism Significance Penicillin Penicillium chrysogenum antibiotic Streptomycin Streptomyces griseus antibiotic Citric acid food industry 30 similarities differences primary vs. secondary metabolites Contents P.M. S.M Growth needed? Yes No Species specificity? No Yes Production stage during active growth after growth phase Examples amino acid, nucleotide, polysaccharide, vitamin antibiotics, toxin, hormones, pigment Produced steadily by cell regulations 31 Steps in Industrial Fermentation Screen the best producing strain Prepare medium and sterilization Optimize production condition Downstream processing 32 raw materials stock culture medium formulation shake flask sterilization seed fermenter fermenter air recovery purification 33 products Outline of a fermentation process Raw Materials Production microorganism Fermentation Upstream processing Downstream processing Product purification 34 Effluent waste Product Fermentation engineering UPSTREAM PROCESSES FERMENTATION Process Control DOWNSTREAM PROCESSES 35 Fermentation engineering UPSTREAM PROCESSES - Producer microorganism - Inoculum development - Media formulation - Sterilization - Fermentation Process FERMENTATION Process Control 36 The essential components of a fermentor 37 Fermentation engineering DOWNSTREAM PROCESSES FERMENTATION Process Control - product extraction, purification & assay - waste treatment -by product recovery The ratio of recovery to fermentation costs for L-asparaginase: 3.0 ethanol: 0.16 38 How are microorganisms cultivated? • Petri Dishes • Shake Flasks • Fermentor 39 The Fermentation Process 40 Industrial production of yeast cells 41 42 Review Questions 1. Cite the following scientists’ outstanding contributions to the fermentation industry: leenvenhoek、Pasteur、Koch and Buchner 2. What is fermentation and what are the main components of fermentation engineering? 3. What are the primary metabolites and the secondary metabolites? Give the differences between them with examples. 43