Propionibacterium acnes, a resident of lipid-rich human skin, produces a 33 kDa extracellular lipase encoded by gehA

Abstract

Five independent clones of the Propionibacterium acnes P-37 lipase gene (gehA) were obtained in ischerichia 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) radiolabelled 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 17 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.

Purification and characterization of an intracellular esterase from Propionibacterium freudenreichii ssp. freudenreichii ITG 14

“…On the contrary, Dupuis and Boyaval [4], and Dupuis et al [5] reported extracellular or cell wall bound lipase and esterase activities for several Propionibacterium strains. Extracellular lipase has been also reported for P. acidipropionici [21], while the existence of extracellular lipases in the case of cutaneous propionic acid bacteria has been clearly confirmed [12,18].…”

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

“…Before the genome decipherment, knowledge of the capacity of P. acnes to use skin tissue was limited to a secreted extracellular triacylglycerol lipase, GehA, isolated from P. acnes P-37 (294). This enzyme degrades skin lipids, such as sebum, which may be a crucial activity for skin coloni- zation.…”

Genetics and molecular biology of propionibacteria

“…The number of genes sequenced from the cutaneous strains of Propionibacterium is even lower, and includes a lipase [28] and a hyaluronate lyase encoding gene (public databases; see Tab. I).…”

“…sherm. [21] lipase gehA P. acnes [28] Plasmid-encoded replication proteins putative rolling circle replicase rep P. freud. pLME108 [3,4] putative theta replicase repA P. acidi.…”