title: Lipase from Chromobacterium viscosum: biochemical characterization indicating homology to the lipase from Pseudomonas glumae.
abstract: Previous purification of a commercial lipolytic preparation from Chromobacterium viscosum using gel filtration chromatography yielded two enzymatically active fractions, named lipases A and B. Characterization of these fractions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that lipase A consisted of a high molecular weight aggregate of lipase protein with lipopolysaccharides. This complex could be dissociated by treatment with EDTA-Tris buffer containing the non-ionic detergent n-octyl-beta-D-glucopyranoside and subsequent isoelectric focusing in an agarose gel containing the same detergent. Both lipases A and B revealed a major peak corresponding to an isoelectric point of 7.1. SDS-PAGE analysis of lipases A and B after purification by gel filtration or by IEF revealed one major protein band of M(r) of 33 K. Determination of N-terminal amino acid sequences confirmed that both fractions A and B contained the same lipase protein. Furthermore, the N-terminal amino acid sequence of the C. viscosum lipase was identical to the one of Pseudomonas glumae lipase.
authors.affiliation: Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, Lisboa, Portugal.
authors.forename: M A
authors.initials: MA
authors.lastname: Taipa
authors.forename: K
authors.initials: K
authors.lastname: Liebeton
authors.forename: J V
authors.initials: JV
authors.lastname: Costa
authors.forename: J M
authors.initials: JM
authors.lastname: Cabral
authors.forename: K E
authors.initials: KE
authors.lastname: Jaeger
mesh_terms: D000595:Amino Acid Sequence; D002861:Chromobacterium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification*; D008070:Lipopolysaccharides / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011549:Pseudomonas / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid
chemical_list: D008070:Lipopolysaccharides; D008049:Lipase