1887

Abstract

SUMMARY: A family of mutant amidases has been derived by experimental evolution of the aliphatic amidase of strain PAC1. Mutation in the structural gene for the enzyme, results in the production of the mutant B amidase by strain B6. This strain, unlike the wild-type, can utilize butyramide for growth. Strain B6 gave rise by a single mutational event to strain V9, utilizing valeramide, and strain PhB3, utilizing phenylacetamide. Strain V9 was not itself able to utilize phenylacetamide but gave rise by mutation to the phenylacetamide-utilizing mutant PhVI. Peptide 108 was isolated from chymotryptic digests of mutant amidases from strains B6, PhB3 and PhV1, but could not be detected in chymotryptic digests of the wild-type amidase. The sequence of peptide 108 was established as Met-Arg-His-Gly-Asp-Ile-Phe. Thermolytic digests of mutant amidases from strains B6, PhB3, PhV1 and V9 were compared with digests of the wild-type amidase. A peptide of the composition Met, Arg, His, Gly Asp Ile, Ser Thr, Val was found in the digest of the wild-type amidase and was replaced in the digests of the mutant amidases by a peptide of the composition Met, Arg, His, Gly Asp Ile, Ser Thr, Val, Phe. Mutation is common to the four mutant enzymes and can be accounted for by the mutation Ser Phe. The sequence of the chymotryptic peptide corresponds with the N-terminal sequence of the amidase protein, and can also be related to the thermolysin peptides. It is concluded that mutation is a Ser the change at position 7 from the N-terminus and the effect of this on the enzyme conformation is discussed.

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/content/journal/micro/10.1099/00221287-114-1-75
1979-09-01
2019-10-20
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http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-114-1-75
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