Fermentation Presentation
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Transcript of Fermentation Presentation
Biotechnology
The integrated use of biochemistry, microbiology and chemical engineering to exploit plant materials and genetic resources for the production of specific products and services
Fermentation
Fermentation is considered as 'any process for the production of a product by means of mass culture of micro-organisms'.
FERMENTATION BIOTECHNOLOGY
MICRO-ORGANISMS
• Several species belonging to the following categories of micro-organisms are used in fermentation processes:
• PROKARYOTICUnicellular : Bacteria, CyanobacteriaMulticellular : Cyanobacteria
• EUKARYOTIC Unicellular : Yeasts, algaeMulticellular : Fungi, algae
REQUIREMENTS FOR ARTIFICIAL CULTURE
Growth of Microorganisms under artificial culture requires Growth (Nutrient) Medium
Microbial Growth is influenced by:
1) Kind and concentration of the ingredients of the medium.
2) pH
3) Temperature
4) Purity of the cultured organism
NUTRIENT SOURCES FOR INDUSTRIAL FERMENTATION
Carbon source
Glucose Corn sugar, Starch, Cellulose
Sucrose Sugarcane, Sugar beet molasses
Lactose Milk whey
Fats Vegetable oils
Hydrocarbons Petroleum fractions
Nitrogen sourceProtein Soybean meal, Corn steep liquor, Distillers' soluble
Ammonia Pure ammonia or ammonium salts
Nitrate Nitrate salts
Phosphorous source Phosphate salts
PHASES OF MICROBIAL GROWTH
Lag phase
Period of adaptation
Log phase (Tophophase or growth phase in case of Fungi)
Rate of growth of the organism steadily increases.
Stationary phase (Idiophase or production phase in case of Fungi)
Growth ceases
FERMENTERS AND BIOREACTORS
A large growth vessel used to culture microorganisms on a large scale, frequently for the production of some commercially valuable product
A bioreactor differs from a fermenter in that the former is used for the mass culture of plant or animal cells, instead of micro-organisms.
Fermenters/bioreactors are equipped with an aerator to supply oxygen in aerobic processes, a stirrer to keep the concentration of the medium uniform, and a thermostat to regulate temperature, a pH detector and similar control devices.
DESIGN OF INDUSTRIAL FERMENTATION PROCESS
• The fermentation process requires the following:
a) a pure culture of the chosen organism, in sufficient quantity and in the correct physiological state;
b) sterilised, carefully composed medium for growth of the organism;
c) a seed fermenter, a mini-model of production fermenter to develop an inoculum to initiate the process in the main fermenter;
d) a production fermenter, the functional large model; and
e) equipment for i) drawing the culture medium in steady state, ii) cell separation, iii) collection of cell free supernatant, iv) product purification, and v) effluent treatment.
Items a) to c) above constitute the upstream and e) constitutes the downstream, of the
fermentation process
FERMENTORS
TYPES OF CULTURE SYSTEMS
BATCH CULTURE
Culture is stopped at stationary phase for recovery.
Advantage- optimum levels of product recovery.
Disadvantages- wastage of unused nutrients, the peaked input of labour and the time lost between batches.
FED-BATCH CULTURE
A fresh aliquot of the medium is continuously or periodically added, without the removal of the culture fluid.
A low but constantly replenished medium has the following advantages:
maintaining conditions in the culture within the aeration capacity of the fermenter;
removing the repressive effects of medium components such as rapidly used
carbon and nitrogen sources and phosphate;
avoiding the toxic effects of a medium component; and
providing limiting level of a required nutrient for an auxotrophic strain.
CONTINUOUS CULTURE
Growth of the organism is controlled by the availability of growth limiting chemical component of the medium
Continuous processing may suffer from contamination, both from within and outside
Continuous culturing is highly selective and favours the propagation of the best-adapted organism in culture
PRODUCTS OF FERMENTATION PROCESSES
Fermentation processes aim at one or more of the following:
Production of cells (biomass) such as yeasts; Single cell protein
Extraction of metabolic products such amino acids, proteins (including enzymes), vitamins, alcohol, etc., for human and/or animal consumption or industrial use such as fertiliser production;
Primary metabolitesSecondary metabolites
Modification of compounds (through the mediation of elicitors or through biotransformation); and
Production of recombinant products.
Genetically manipulated Escherichia coli, Saccharomyces cerevisiae, other yeasts and even filamentous fungi are now being used to produce
interferon, insulin, human serum albumin, and several other products.
GENETIC IMPROVEMENT OF FERMENTATION PROCESSES
MUTATION
Exposing a culture of a micro-organism to UV light, ionising radiation or certain chemicals
RECOMBINATION
Recombination is defined as any process that brings together genes from different sources
DNA MANIPULATION
INDUSTRIAL FERMENTATION
Microbiology lab
Cell maintenance and storage Preparation of master and working cell banks Inoculum Development Sterility and purity checking of fermenter samples
Fermentation
Media Preparation tanks Seed Tanks Fermentors Dosing/Feed tanks Online control – DCS system
Down Stream Processing
Centrifuges/Filters solvent solvent extraction vessel and layer separators Chromatographic columns Dryers Sieve's
QC/QA and Analytical Development Lab
Analysis of in process samples Analysis of raw materials Finished good analysis Impurity profiling Analytical method development Microbiological testing of finished goods and process water. Environmental monitoring of the plant QA functions
Engineering
Supply of utilities like Steam, instrument air, cooling water, chilled water etc. Maintenance works Validation and calibration of equipments/instruments
Store/Warehouse
Storage of raw materials Storage of finished goods Storage of engineering spares Storage of solvents and gas cylinders
Microbiology lab - Inoculum Development
Master cell bank (Lyophil vials)
Working cell bank
(slants or glycerol stocks)
Spore or vegetative cell suspension as inoculum
Slants or shake flask
Fermentation
Seed
Seed media preparation and sterilization. Inoculation of seed tanks with laboratory inoculum aseptically. Growing seed culture (15-48 hrs). Control of pH, air and agitation. Sampling and microbiological testing
Production
Production media preparation and sterilization. Inoculation of production fermenter with appropriately grown seed aseptically. Running of fermentation under controlled conditions ( pH, dissolved oxygen, agitation
and feed rate). Sampling and microbiological testing Run time (1 day to 15 days)
Harvest
Down Stream Processing
Fermentation broth storage
Cell disruption- Sonication, Beads, French press, Microfluidizer ect.
Cell/cell debries separation- Press filters, RVFs, Nutch Filters, Centrifuges etc.
Purification of product by solvent solvent extraction or chromatography
Precipitation/Lyophilization
Drying
Packaging
OPERATIONAL DIAGRAM OF LARGE-SCALE FUNGAL BATCH FERMENTATION SYSTEM
Preculture Preparation of Fermentation Recovery of enzyme-inoculum containing medium
FERMENTATION PLANT OPERATIONS
Microbiology Laboratory
Fermentation Production
Down Stream Processing/Recovery
Quality Control/QA
Store/Warehouse
Effluent Treatment/Incineration
API