The Pseudomonas aeruginosa acsA gene, encoding an acetyl-CoA synthetase, is essential for growth on ethanol.
Pseudomonas aeruginosa ATCC 17933 uses a pyrroloquinoline quinone-dependent ethanol oxidation system. Two mutants of P. aeruginosa, 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 acsA 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 acsA gene restored growth and led to the expression of ACS activity in excess of that of wild-type cells. In wild-type P. aeruginosa, 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 P. aeruginosa requires active acsA gene product for growth on ethanol.
Kretzschmar
Utta
U
Fachgebiet Technische Biochemie, Institut für Biotechnologie der Technischen Universität Berlin, Seestraße 13, D-13353 Berlin, Germany1.
Schobert
Max
M
Fachgebiet Technische Biochemie, Institut für Biotechnologie der Technischen Universität Berlin, Seestraße 13, D-13353 Berlin, Germany1.
Görisch
Helmut
H
Fachgebiet Technische Biochemie, Institut für Biotechnologie der Technischen Universität Berlin, Seestraße 13, D-13353 Berlin, Germany1.
D000085:Acetates / Q000378:metabolism; D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism; D003001:Cloning, Molecular; D003470:Culture Media; D000431:Ethanol / Q000378:metabolism*; D017353:Gene Deletion; D005798:Genes, Bacterial; D020043:Genes, Essential; D005816:Genetic Complementation Test; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*
D000085:Acetates; D003470:Culture Media; D009097:Multienzyme Complexes; D000431:Ethanol; D002244:Carbon; D000445:Aldehyde Oxidoreductases; C019823:carbon monoxide dehydrogenase