Purification of an epoxide hydrolase from Rhodotorula glutinis

Citation
Nae. Kronenburg et al., Purification of an epoxide hydrolase from Rhodotorula glutinis, BIOTECH LET, 21(6), 1999, pp. 519-524
Citations number
20
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
BIOTECHNOLOGY LETTERS
ISSN journal
01415492 → ACNP
Volume
21
Issue
6
Year of publication
1999
Pages
519 - 524
Database
ISI
SICI code
0141-5492(199906)21:6<519:POAEHF>2.0.ZU;2-H
Abstract
The epoxide hydrolase from Rhodotorula glutinis was isolated and initially characterized. The enzyme was membrane associated and could be solubilized by Triton X-100. Purification yielded an enzyme with sp. act. of 66 mu mol 1,2-epoxyhexane hydrolyzed min(-1) mg(-1) protein. The enzyme was not compl etely purified to homogeneity but, nevertheless, a major protein was isolat ed by SDS-PAGE for subsequential amino acid determination of peptide fragme nts. From sequence alignments to related enzymes, a high homology towards t he active site sequences of other microsomal epoxide hydrolases was found. Molecular mass determinations indicated that the native enzyme exists as a homodimer, with a subunit molecular mass of about 45 kDa. Based upon these, this epoxide hydrolase is structurally related to other microsomal epoxide hydrolases.