f Propionibacterium acnes, a resident of lipid-rich human skin, produces a 33 kDa extracellular lipase encoded by gehA
- Authors: James E. Miskin, Angela M. Farrell, William J. Cunliffe, Keith T. Holland
- Author for correspondence: Keith T. Holland. Tel: + 44 113 2335647. Fax: +44 113 2335638.
- Microbiology, May 1997 143: 1745-1755, doi: 10.1099/00221287-143-5-1745
- Subject: Genetics And Molecular Biology
- Published Online:
Five independent clones of the Propionibacterium acnes P-37 lipase gene (gehA) were obtained in Escherichia coli, and the gene was localized to a 2.75 kb Xhol fragment by subcloning. The five clones were shown to contain the same gene by Southern blotting with a DIG-labelled probe to gehA. The nucleotide sequence of gehA was determined, and shown to contain a single ORF of 1017 kb, encoding a protein of 339 amino acids. The predicted molecular mass was 36 kDa. A 33 kDa (PAGE) radiolabeled polypeptide was detected from E. coli minicell preparations harbouring gehA, which could correspond to GehA after cleavage of the putative 26 amino acid residue signal peptide. gehA was overexpressed in E. coli under the control of the bacteriophage T7 promoter, and the corresponding polypeptide was found to be present in insoluble aggregates. Active lipase was produced when the overexpressing strain was incubated at a reduced temperature in the presence of sucrose. Purification of lipase from P. acnes culture supernatant fluids confirmed the production of a 33 kDa (PAGE) lipase.
© Society Society for General Microbiology 1997 | Published by the Microbiology Society
Article metrics loading...
Full text loading...
Author and Article Information
/content/journal/micro/10.1099/00221287-143-5-1745dcterms_title,dcterms_subject,pub_serialTitlepub_serialIdent:journal/micro AND -contentType:BlogPost64
/content/journal/micro/10.1099/00221287-143-5-1745dcterms_title,dcterms_subject-pub_serialIdent:journal/micro AND -contentType:BlogPost64
Figure data loading....