1887

Abstract

ATCC 17933 uses a pyrroloquinoline quinone-dependent ethanol oxidation system. Two mutants of , unable to grow on ethanol and showing no acetyl-CoA synthetase (ACS) activity under standard test conditions, were complemented by cosmid pTB3018. Subcloning led to the isolation of a gene which encodes a protein with high similarity to acetyl-CoA synthetases. Interruption of the putative gene by a kanamycin-resistance cassette resulted in a mutant also unable to grow on ethanol and with very low residual acetyl-CoA-forming activity. Complementation by the wild-type allele of the gene restored growth and led to the expression of ACS activity in excess of that of wild-type cells. In wild-type , ACS activity was induced upon growth on ethanol, 2,3-butanediol, malonate and acetate. The wild-type and mutants defective in ACS activity showed an active acetate kinase (ACK) under the growth conditions used; however, phosphotransacetylase (PTA) could not be detected. The data indicate that requires active gene product for growth on ethanol.

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2001-10-01
2019-10-21
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