Fed batch

Commercial-scale operations are conducted in batch, fed-batch, or continuous culture systems. Fermentation vessels include the conventional baffled aerated tank, with or without impeller agitation, and the ak-lift tower fermentors in which ak is sparged into an annular space between the... 

S. cerevisiae is produced by fed-batch processes in which molasses supplemented with sources of nitrogen and phosphoms, such as ammonia, ammonium sulfate, ammonium phosphate, and phosphoric acid, are fed incrementally to meet nutritional requirements of the yeast during growth. Large (150 to 300 m ) total volume aerated fermentors provided with internal coils for cooling water are employed in these processes (5). Substrates and nutrients ate sterilized in a heat exchanger and then fed to a cleaned—sanitized fermentor to minimize contamination problems. 

C. uti/is yeast is produced by either fed-batch or continuous processes. Aerated-agitated fermentors range up to 300 m total capacity and ate operated in the same manner as described for S. cerevisiae (2,5). C. utilis is capable of metabolizing both hexose and pentose sugars. Consequendy, papermiU wastes such as sulfite waste Hquot that contain these sugars often ate used as substrates. 

The batch and fed-batch procedures are used for most commercial antibiotic fermentations. A typical batch fermentor may hold over 150,000 Hters. When a maximum yield of antibiotic is obtained, the fermentation broth is processed by purification procedures tailored for the specific antibiotic being produced. Nonpolar antibiotics are usually purified by solvent extraction procedures water-soluble compounds are commonly purified by ion-exchange methods. Chromatography procedures can readily provide high quaHty material, but for economic reasons chromatography steps are avoided if possible. 

Bakers Yeast Production. Bakers yeast is grown aerobicaHy in fed-batch fermentors under conditions of carbohydrate limitation. This maximizes the yield of yeast biomass and minimizes the production of ethanol. Yeasts grown under these conditions have exceUent dough leavening capabHity and perform much better in the bakery than yeast grown under anaerobic conditions. 

Because of the differences in primary and secondaiy metabolism, a reactor may have a dual-stage fed-batch system. In other words, fed-batch operation optimizes growth with little or no product formation. When sufficient biomass has accumulated, a different fed-batch protocol comes into play. 

Fed-batch culture A cell cultivation technique in which one or more nutrients are supplied to the bioreactor in a given sequence during the growth or bioconversion process while the products remain in the vessel until the end of the run. 

Keller, R. and Dunn, I. J., Computer simulation of the biomass production rate of cyclic fed batch continuous culture, J. Appl. Chem., BiotechnoL, 28, 508-514, 1978. 

There are three idealized flow reactors fed-batch or semibatch, continuously stirred tank, and the plug flow tubular. Each of these is pictured in Figure 1. The fed-batch and continuously stirred reactors are both taken as being well mixed. This means that there is no spatial dependence in the concentration variables for each of the components. At any point within the reactor, each component has the same concentration as it does anywhere else. The consequence... 

Biocatalysts in nature tend to be optimized to perform best in aqueous environments, at neutral pH, temperatures below 40 °C, and at low osmotic pressure. These conditions are sometimes in conflict with the need of the chemist or process engineer to optimize a reaction with respect to space-time yield or high product concentration in order to facilitate downstream processing. Furthermore, enzymes and whole cells are often inhibited by products or substrates. This might be overcome by the use of continuously operated stirred tank reactors, fed-batch reactors, or reactors with in situ product removal [14, 15]. The addition of organic solvents to increase the solubility of substrates and/or products is a common practice [16]. 

As you might have already gathered, the majority of industrial fermentations are batch processes. In closed batch systems, the growth medium is inoculated with cells and growth and product formation is allowed to proceed until the required amount of conversion has taken place. After harvesting the culture the vessel is cleaned, sterilised and filled with fresh medium prior to inoculation. For some processes, addition of all the feedstock prior to inoculation, as is done in closed batch fermentations, is undesirable and it is preferable to incrementally add the carbon source as the fermentation proceeds. Such a process is known as fed-batch culture and the approach is often used to extend the lifetime of batch cultures and thus product yields fed-batch cultures are considered further in Section 2.7.4. 

In many cases, problems cannot be overcome by biological means. This is especially true for those related to inhibition by substrate or product. There may, however, be technical solutions to these problems. Nowadays, complicated feed strategies with different substrates can be achieved through the use of flow injection analysis, on-line sensors, mass flow meters and sophisticated computer control. Such control coupled to a fed-batch mode of operation (Figure 2.5) can often eleviate problems caused by substrate inhibition. For some processes, continuous product removal can avoid the problems associated with product inhibition the various options include ... 

Figure 2.5 Possible technological solutions to bioprocess problems a) Fed-batch culture b) Continuous product removal (eg dialysis, vacuum fermentation, solvent extraction, ion exchange etc) c) Two-phase system combined with extractive fermentation (liquid-impelled loop reactor) d) Continuous culture, internal multi-stage reactor e) Continuous culture, dual-stream multi-stage reactor f) Continuous culture with biomass feedback (cell recycling). (See text for further details).

When excess substrate interferes with growth and/or product formation. One example is the production of baker s yeast. It is known that relatively low concentrations of certain sugars repress respiration and this will make the yeast cells switch to fermentative metabolism, even under aerobic conditions. This, of course, has a negative effect on biomass yield. When maximum biomass production is aimed at, fed batch cultures are the best choice, since the concentration of limiting sugar remains low enough to avoid repression of respiration. 

Process A Genetic instability Substrate repression Multi-step synthesis Product (volatile) inhibition Mode of operation Batch Fed-batch Continuous... 

Fed-batch fermentations are batch fermentations which are fed continuously, or intermitantly, with medium without the removal of fluid. In this way the volume of the culture increases with time. 

One of the advantages of the fed-batch fermentation is the fact that the residual substrate concentration may be maintained at a very low level. This may result in a removal of catabolite repressive effects and avoidance of toxic effects of medium components. 

Cell production can be carried out by a normal fed-batch type of fermentation. The feed rate of glucose is increased during the fermentation and the cells grow exponentially. 

The culture can be used directly for the conversion of phenylpyruvic add to resting cells L-phenylalanine. Therefore, a batch process with resting cells can be carried out, with some glucose added for maintenance (fed-batch fermentation). Another approach is to harvest the cells from the fermentation broth and to use them in a separate bioreactor in higher concentrations than the ones obtained in the cell cultivation. An advantage of the last method can be that the concentration of compounds other than L-phenylalanine is lower, so that downstream processing may be cheaper. 

Figure 8.7 Fed-batch fermentation of phenylpyruvic acid to L-phenylalanine.

In fed-batch mode residual substrate concentration may be maintained at very low levels. Ibis would reduce substrate costs, may remove catabolic repressive effects and may avoid possible toxic effects of the substrate. The fed-batch mode of operation may also avoid oxygen depletion of the culture during rapid growth. 

A full set of bioreactors with pH and temperature controllers are shown in Figure 1.3. The complete set of a 25 litre fermenter with all the accessory controlling units creates a good opportunity to control suitable production of biochemical products with variation of process parameters. Pumping fresh nutrients and operating in batch, fed batch and continuous mode are easy and suitable for producing fine chemicals, amino acids, and even antibiotics. 

Fed batch is used to overcome substrate limitations, especially for the production of antibiotics. 

Fed-batch mixed reactor starting with a relatively dilute solution of substrate this provides control over the substrate concentration. High rates are avoided. Fed batch is used for baker s yeast to overcome catabolite repression and to control oxygen demand. It is also used routinely for production of Penicillin. 

Batch mixed reactor There are three principal modes of bioreactor operation (a) batch (b) fed batch (c) continuous. 

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