Abstract
A 2.9-kb SacI fragment containing the ask-asd operon, encoding aspartokinase and aspartate-semialdehyde dehydrogenase, was cloned from an aminoethylcysteine-resistant, lysine-producing Corynebacterium lactofermentum strain. Enzymatic analysis showed that the aspartokinase (ASK) activity was completely resistant to inhibition by mixtures of lysine and threonine. Comparison of the deduced amino acid sequence of the β submit of the ask gene showed three amino acid residue changes with ask genes encoding wild-type, feedback-sensitive enzymes. Three C. lactofermentum strains, one being aspartokinase-negative, one carrying two ask genes on the chromosome and one having a sixfold higher specific ASK activity than the parental strain, were constructed by transconjugation and electroporation, and used to analyse the role of ASK in the lysine production by C. lactofermentum. The results indicate that, in this study, feed-back-resistant ASK is necessary for high-level lysine production, but dispensable for lysine and diaminopimelate synthesis required for cell growth.
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Jetten, M.S.M., Follettie, M.T. & Sinskey, A.J. Effect of different levels of aspartokinase of the lysine production by Corynebacterium lactofermentum . Appl Microbiol Biotechnol 43, 76–82 (1995). https://doi.org/10.1007/BF00170626
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DOI: https://doi.org/10.1007/BF00170626