1887

Abstract

PP3 carrying dehalogenases I and II and PAU3 carrying dehalogenase I coded for by plasmid pUU2 were able to grow on 2-monochloropropionic acid (2MCPA). Neither strain utilized 2-chloropropionamide (2CPA) as a carbon or nitrogen source for growth. Mutations in both strains to 2Cpa phenotypes (designated PPW3 and PAU5, respectively) involved the expression of an acquired 2CPA-amidase activity. The amidase followed by dehalogenase reactions in these strains constituted a novel metabolic pathway for growth on 2CPA. PPW3 synthesized a constitutive amidase of molecular mass 59 kDa consisting of two identical subunits of 29 kDa. For those amides tested this acquired enzyme was most active against chlorinated aliphatic amides, although substrate affinities ( ) and maximum rates of activity ( ) were poor. PAU5 acquired a 2Cpa phenotype by overproducing the A-amidase normally used by this species to hydrolyse aliphatic amides. The A-amidase had only slight activity towards 2CPA. However, with constitutive synthesis the mutant grew on the chlorinated substrates. Chloroacetamide (CAA) was a toxic substrate analogue for these strains. A strain resistant to CAA was isolated from PAU5 when exposed to 1-10 mM-CAA. This mutant, PAU6, synthesized an inducible A-amidase. CAA-resistance depended upon the simultaneous expression of CAA-inducible amidase and dehalogenase activities.

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/content/journal/micro/10.1099/00221287-132-8-2195
1986-08-01
2019-10-14
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http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-132-8-2195
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