1887

Abstract

The lysine biosynthetic pathway has to supply large amounts of -aminoadipic acid for penicillin biosynthesis in . In this study, we have characterized the L2 mutant, a lysine auxotroph that shows highly increased expression of several lysine biosynthesis genes (, , , ). The L2 mutant was found to be deficient in homoaconitase activity since it was complemented by the gene. We have cloned a gene (named ) that complements the L2 mutation by transformation with a genomic library, constructed in an autonomous replicating plasmid. The -encoded protein showed high identity to homoaconitases. In addition, we cloned the mutant allele from the L2 strain that showed a G to A point mutation resulting in a Gly to Asp substitution. This mutation is located in a highly conserved region adjacent to two of the three cysteine residues that act as ligands to bind the iron–sulfur cluster required for homoaconitase activity. The L2 mutant accumulates homocitrate. Deletion of the gene (homocitrate synthase) in the L2 strain prevented homocitrate accumulation and reverted expression levels of the four lysine biosynthesis genes tested to those of the parental prototrophic strain. Homocitrate accumulation seems to act as a sensor of lysine-pathway distress, triggering overexpression of four of the lysine biosynthesis genes.

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2009-12-01
2020-03-31
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