Introduction

Detailed investigation is needed to establish the most suitable medium for an individual fermentation process, but certain basic requirements must be met by any such medium. All micro-organisms require water, sources of energy, carbon, nitrogen, mineral elements and possibly vitamins plus oxygen if aerobic. On a small scale it is relatively simple to devise a medium containing pure compounds, but the resulting medium, although supporting satisfactory growth, may be unsuitable for use in a large scale process.

On a large scale one must normally use sources of nutrients to create a medium which will meet as many as possible of the following criteria:

1. It will produce the maximum yield of product or biomass per gram of substrate used.

2. It will produce the maximum concentration of product or biomass.

3. It will permit the maximum rate of product formation.

4. There will be the minimum yield of undesired products.

5. It will be of a consistent quality and be readily available throughout the year.

6. It will cause minimal problems during media making and sterilization.

7. It will cause minimal problems in other aspects of the production process particularly aeration and agitation, extraction, purification and waste treatment.

The use of cane molasses, beet molasses, cereal grains, starch, glucose, sucrose and lactose as carbon sources, and ammonium salts, urea, nitrates, corn steep liquor, soya bean meal, slaughter-house waste and fer mentation residues as nitrogen sources, have tended to meet most of the above criteria for production media because they are cheap substrates. However, other more expensive pure substrates may be chosen if the overall cost of the complete process can be reduced because it is possible to use simpler procedures. Other criteria are used to select suitable sporulation and inoculation media and these are considered in Chapter 6.

It must be remembered that the medium selected will affect the design of fermenter to be used. For example, the decision to use methanol and ammonia in the single cell protein process developed by ICI pic necessitated the design of a novel fermenter design (MacLennan et al., 1973; Sharp, 1989). The microbial oxidation of hydrocarbons is a highly aerobic and exothermic process. Thus, the production fermenter had to have a very high oxygen transfer capacity coupled with excellent cooling facilities. ICI pic solved these problems by developing an air lift fermenter (see Chapter 7). Equally, if a fermenter is already available this will obviously influence the composition of the medium. Rhodes et al. (1955) observed that the optimum concentrations of available nitrogen for griseofulvin production showed some variation with the type of fermenter used. Some aspects of this topic are considered in Chapter 7.

The problem of developing a process from the laboratory to the pilot scale, and subsequently to the industrial scale, must also be considered. A laboratory medium may not be ideal in a large fermenter with a low gas-transfer pattern. A medium with a high viscosity will also need a higher power input for effective stirring. Besides meeting requirements for growth and product formation, the medium may also influence pH variation, foam formation, the oxidation-reduction potential, and the morphological form of the organism. It may also be necessary to provide precursors or metabolic inhibitors. The medium will also affect product recovery and effluent treatment.

Historically, undefined complex natural materials have been used in fermentation processes because they are much cheaper than pure substrates. However, there is often considerable batch variation because of variable concentrations of the component parts and impurities in natural materials which cause unpredictable biomass and/or product yields. As a consequence of these variations in composition small yield improvements are difficult to detect. Undefined media often make product recovery and effluent treatment more problematical because not all the components of a complex nutrient source will be consumed by the organism. The residual components may interfere with recovery (Chapter 10) and contribute to the BOD of the effluent (Chapter 11).

Thus, although manufacturers have been reluctant to use defined media components because they are more expensive, pure substrates give more predictable yields from batch to batch and recovery, purification and effluent treatment are much simpler and therefore cheaper. Process improvements are also easier to detect when pure substrates are used.

Collins (1990) has given an excellent example of a process producing recombinant protein from S. cere-uisiae instead of just biomass. The range of growth conditions which can be used is restricted because of factors affecting the stability of the recombinant protein. The control of pH and foam during growth in a fermenter were indentified as two important parameters. Molasses would normally be used as the cheapest carbohydrate to grow yeast biomass in a large scale process. However, this is not acceptable for the recombinant protein production because of the difficulties, and incurred costs caused in subsequent purification which result from using crude undefined media components. Collins and co-workers therefore used a defined medium with glucose, sucrose or another suitable carbon source of reasonable purity plus minimal salts, trace elements, pure vitamins and ammonia as the main nitrogen source and for pH control. Other impurities in molasses might have helped to stabilize foams and led to the need to use antifoams.

Aspects of microbial media have also been reviewed by Suomalainen and Oura (1971), Martin and Demain (1978), Iwai and Omura (1982), DeTilly et al. (1983), Kuenzi and Auden (1983), Miller and Churchill (1986), Smith (1986) and Priest and Sharp (1989).

Media for culture of animal cells will be discussed later in this chapter.

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