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FERMENTATION Aerobic Anaerobic Adequate aeration In addition, these fermenters may have a mechanism for stirring and mixing of the medium and cells Eg: Antibiotics, enzymes, vitamins In anaerobic fermentation, a provision for aeration is usually not needed. Eg: Lactic acid, ethanol, wine Fermentation could be: Batch Continuous Most fermentations are batch processes. Nutrients and the inoculum are added to the sterile fermenter and left to get on with it. Anti-foaming agent may be added. Once the desired amount of product is present in the fermenter the contents are drained off and the product is extracted. After emptying, the tank is cleaned & prepared for a new batch. Some products are made by a continuous culture system. Sterile medium is added to the fermentation with a balancing withdrawal of broth for product extraction. Fed-batch fermentation is a production technique in between batch and continuous fermentation. A proper feed rate, with the right component constitution is required during the process. Fed-batch offers many advantages over batch and continuous cultures. What it is? In contrast to Submerged (liquid state) Fermentation, Solid State Fermentation (SSF) is the cultivation of micro organisms under controlled conditions in the absence of free water. Examples of products of Solid State Fermentation include industrial enzymes, fuels and nutrient enriched animal feeds. The application of modern biotechnical knowledge and process control technologies can lead to significant productivity increases from this ancient process. Higher volumetric productivity Usually simpler with lower energy requirements Might be easier to meet aeration requirements Resembles the natural habitat of some fungi and bacteria Easier downstream processing Techniques for large-scale production of microbial products. It must both provide an optimum environment for the microbial synthesis of the desired product and be economically feasible on a large scale. They can be divided into surface (emersion) and submersion techniques. The latter may be run in batch, fed batch, continuous reactors. In the surface techniques, the microorganisms are cultivated on the surface of a liquid or solid substrate. These techniques are very complicated and rarely used in industry. Upstream processing encompasses any technology that leads to the synthesis of a product. Upstream includes the exploration, development and production. • The media is the feed solution ◦ It must contain the essential nutrients needed for the microbe to grow • Factors of consideration when choosing media -Quality consistence and availability -Ensure there are no problems with Media Prep or other aspects of production process Ex. Cane molasses, beet molasses, cereal grains • • The type: complex and synthetic medium (mineral medium) Even small modifications in the medium could change cell line stability, product quality, yield, operational parameters, and downstream processing. • • • Fermentation medium consists of: Macronutrients (C, H, N, S, P, Mg sources water, sugars, lipid, amino acids, salt minerals) Micronutrients (trace elements/ metals, vitamins) Additional factors: growth factors, attachment proteins, transport proteins, etc) For aerobic culture, oxygen is sparged. • CARBON SOURCES: Lactose, glucose & sucrose may be used. • NITROGEN SOURCES: Corn steep liquor (CSL), ammonium sulphate and ammonium acetate can be used as nitrogenous sources. MINERAL SOURCES: Elements namely potassium, phosphorus, magnesium, sulphur, zinc and copper are essential for penicillin production. Some of these are applied by corn steep liquor. • Calcium can be added in the form of chalk to counter the natural acidity of some media. Incoculum is the substance/ cell culture that is introduced to the medium. The cell then grow in the medium, conducting metabolisms. Inoculum is prepared for the inoculation before the fermentation starts. It needs to be optimized for better performance • The inoculum is the starter culture that is injected into the fermenter ◦ It must be of sufficient size for optimal growth kinetics • Since the production fermenter in industrial fermentations is so large, the inoculum volume has to be quite large - A seed fermenter is usually required to produce the inoculum volume -The seed fermenter’s purpose is not to produce product but to prepare inoculum