Screening and characterization of trehalose-oleate hydrolyzing lipase

Citation
R. Ishimoto et al., Screening and characterization of trehalose-oleate hydrolyzing lipase, FEMS MICROB, 195(2), 2001, pp. 231-235
Citations number
14
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Microbiology
Journal title
FEMS MICROBIOLOGY LETTERS
ISSN journal
0378-1097 → ACNP
Volume
195
Issue
2
Year of publication
2001
Pages
231 - 235
Database
ISI
SICI code
0378-1097(20010220)195:2<231:SACOTH>2.0.ZU;2-G
Abstract
Various soil samples were collected to screen the presence of microorganism s which have ability to degrade TOE. One strain (AKU-883) with good TOE deg rading activity was isolated and identified as Burkholderia cepacia and the extracellular enzyme was purified to homogeneity. The purification was ach ieved by ultrafiltration. Super Q anion-exchange chromatography and Superde x 200HR gel-filtration in the presence of Triton X. The enzyme was purified to 85-fold. and specific activity of 4.910 kU mg protein(-1). The peak pre paration on gel filtration showed a single band of 34 kDa on SDS-PAGE and n ative PAGE which indicate the monomeric nature of the enzyme. The pr of the enzyme was 6.3. The enzyme showed the maximum activity at pH 9 and 65 degr eesC. and was stable in the range of pH 5-10 and up to 60 degreesC. Almost all the activity (92%) was kept after incubation for more than 1 week at 50 degreesC (pH 7.3). High activities remained even in water-miscible solvent s such as ethanol, dimethyl formamide, diisopropyl ether, and dioxane, The N-terminal 16 amino acid residues were determined as A-N-G-Y-A-A-T-R-Y-P-I- I-L-V-G-G. which showed a consensus sequence for lipases from Burkholderia species. Thus the enzyme was concluded to be a kind of lipase. (C) 2001 Fed eration of European Microbiological Societies. Published by Elsevier Scienc e B.V. All rights reserved.