Purification and characterization of tert-butyl ester-hydrolyzing lipase from burkholderia sp. YY62

13Citations
Citations of this article
6Readers
Mendeley users who have this article in their library.

This article is free to access.

Abstract

An intracellular novel lipase which can hydrolyze t-butyl octanoate (TBO) was purified to homogeneity from crude cell-free extracts of Burkholderia (formerly Pseudomonas) sp. YY62 with 9% overall yield. Seventy-four-fold purification was achieved by ammonium-sulfate precipitation, three consecutive open-column chromatographies (DEAE anion-exchange, Sepharose CL-6B gel-filtration, and the second DEAE anion-exchange columns), and two HPLCs (TSK G2000SWXL gel-filtration and phenyl 5PW hydrophobic interaction columns). Enzymes hydrolyzing p-nitrophenyl acetate were separated into two peaks (peak I and II) on the hydrophobic HPLC, and only peak II was found to have TBO-hydrolyzing activity. The peak preparation showed a single band of 40 kDa on SDS-PAGE and a molecular mass of 39 kDa on gel-filtration under non-denatured conditions, indicating the monomeric nature of the TBO-hydrolyzing lipase. The lipase showed maximum activity at pH 7.0 and 28°C. The N-terminal 15 amino acid residues were determined as Met-Asp-Phe-Tyr-Asp-Ala-Asn-Glu-Thr-Arg-His-Pro-Glu-Gln-Arg, which showed no homology to known proteins, suggesting that the purified enzyme may belong to a novel class of hydrolase. © 1998, Taylor & Francis Group, LLC. All rights reserved.

Cite

CITATION STYLE

APA

Yeo, S. H., Nihira, T., & Yamada, Y. (1998). Purification and characterization of tert-butyl ester-hydrolyzing lipase from burkholderia sp. YY62. Bioscience, Biotechnology and Biochemistry, 62(12), 2312–2317. https://doi.org/10.1271/bbb.62.2312

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free