Screening and characterization of trehalose-oleate hydrolyzing lipase

Ryo Ishimoto, Manabu Sugimoto, Fusako Kawai

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Various soil samples were collected to screen the presence of microorganisms which have ability to degrade TOE. One strain (AKU-883) with good TOE degrading activity was isolated and identified as Burkholderia cepacia and the extracellular enzyme was purified to homogeneity. The purification was achieved by ultrafiltration, Super Q anion-exchange chromatography and Superdex 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 preparation on gel filtration showed a single band of 34 kDa on SDS-PAGE and native PAGE which indicate the monomeric nature of the enzyme. The pI of the enzyme was 6.3. The enzyme showed the maximum activity at pH 9 and 65°C, and was stable in the range of pH 5-10 and up to 60°C. Almost all the activity (92%) was kept after incubation for more than 1 week at 50°C (pH 7.3). High activities remained even in water-miscible solvents 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.

Original languageEnglish
Pages (from-to)231-235
Number of pages5
JournalFEMS Microbiology Letters
Volume195
Issue number2
DOIs
Publication statusPublished - Feb 20 2001

Fingerprint

Trehalose
Oleic Acid
Lipase
Enzymes
Gel Chromatography
Burkholderia
Burkholderia cepacia
Native Polyacrylamide Gel Electrophoresis
Consensus Sequence
Ultrafiltration
Anions
Chromatography
Polyacrylamide Gel Electrophoresis
Ethanol
Soil
Amino Acids
Water

Keywords

  • Burkholderia species
  • Lipase
  • Trehalose-oleate

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Screening and characterization of trehalose-oleate hydrolyzing lipase. / Ishimoto, Ryo; Sugimoto, Manabu; Kawai, Fusako.

In: FEMS Microbiology Letters, Vol. 195, No. 2, 20.02.2001, p. 231-235.

Research output: Contribution to journalArticle

Ishimoto, Ryo ; Sugimoto, Manabu ; Kawai, Fusako. / Screening and characterization of trehalose-oleate hydrolyzing lipase. In: FEMS Microbiology Letters. 2001 ; Vol. 195, No. 2. pp. 231-235.
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