The Development of Industrial Microorganisms

Introduction
Naturally a wild-type bacterium or yeast cell has become closely adapted through natural selection to its
environment and to competition with other species. The industrial strain produces the desired commercial
metabolite in vast quantities relative to its other metabolic end products. The wild type organism isolated from
natural environment, besides optimizing the cultural condition, requires genetic programming to make a normal
metabolic product in an amount required at commercial level. Such a requirement calls for changing its genetic
information in a way that eliminates undesirable properties or introduces entirely new ones. Such an attempt
may result into shunting of more carbon and possibly other metabolites through metabolic sequences to a
desirable end product formation. The genetic improvement of industrial strains may be achieved by mutation
and/or recombination.
11.1
Mutation
Mutations can be induced in the laboratory by exposing the culture to a mutagenic agents i.e., ionizing
radiations, ultra violet light, nitr'ous acid, nitrosoguanidine, purine and pyrimidine analogues and other mutagenic agents. Mutagens hit genes at random. Each agent may act on
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particular bases or group of bases (i.e., UV-radiation tends to link two adjacent thymines on the same strand of
DNA), but all genes are chains of same bases. It is therefore usually impossible to cause a particular gene to
mutate preferentially, even though, in any gene particular treatment tend to cause mutations primarily at certain
positions. The development of strain by mutation depends on the sensitive test that make it possible to identify
the mutant with a desirable properties. Exposure of culture to point mutagenic agents results into survivors of
small numbers, and very small proportion of which may be efficient producers. It is very difficult to determine
the gene that will be affected by the mutagenic agent as the particular mutation occurs in a very large number of
genes.
The selection of mutants producing high yield of primary / metabolites

Amino acids are primary metabolites, most commonly produced by Corynebacterium glutamicum and
Bravibacterium flavum. Lysine is an important amino acid produced by Corynebacterium glutamicum and
Bravibacterium flavum. In these wild type organisms, the production of lysine is regulated in such a way that, it
is produced to a level to meet the needs. The fermentation is based on understanding the pathways leading to
bacterial biosynthesis of the amino acid and the pathways genetic regulation. In these bacteria lysine is one end
product of a branched pathway that also leads to the synthesis of enough of these amino acids methionine and
threonine.
The regulatory mechanisms involved are the inhibition of the activity of key enzymes in the biosynthetic route
and the repression of their synthesis by the end products when present in the cell at required concentration to meet the
organisms need. Such mechanisms are known as feedback control. That is, accumulation of these two amino acids in
excess of the organisms requirements tend to inhibit their synthesis by inhibiting the enzymes activity; conversely, a
shortage of threonine or lysine increases their rate of synthesis.
Overproduction of lysine (Fig 11. 1) far beyond the bacteriums own needs, has been achieved by isolating two
kinds of mutant. In one kind a mutation in the gene coding for the enzyme homoserine dehydrogenase abolishes the
enzyme activity and thereby prevents the bacterium from making threonine (one of the inhibitory products) and
methionine. In this pathway lysine does not exert any control over the biosynthetic route from aspartic semialdehyde
to lysine. A mutant which could not V catalyse the conversion of aspartic semialdehyde to homoserine would