RRC ID 72261
Author Nishida Y, Adachi K, Kasai H, Shizuri Y, Shindo K, Sawabe A, Komemushi S, Miki W, Misawa N.
Title Elucidation of a carotenoid biosynthesis gene cluster encoding a novel enzyme, 2,2'-beta-hydroxylase, from Brevundimonas sp. strain SD212 and combinatorial biosynthesis of new or rare xanthophylls.
Journal Appl Environ Microbiol
Abstract A carotenoid biosynthesis gene cluster mediating the production of 2-hydroxyastaxanthin was isolated from the marine bacterium Brevundimonas sp. strain SD212 by using a common crtI sequence as the probe DNA. A sequence analysis revealed this cluster to contain 12 open reading frames (ORFs), including the 7 known genes, crtW, crtY, crtI, crtB, crtE, idi, and crtZ. The individual ORFs were functionally analyzed by complementation studies using Escherichia coli that accumulated various carotenoid precursors due to the presence of other bacterial crt genes. In addition to functionally identifying the known crt genes, we found that one (ORF11, named crtG) coded for a novel enzyme, carotenoid 2,2'-beta-hydroxylase, which showed intriguingly partial homology with animal sterol-C5-desaturase. When this crtG gene was introduced into E. coli accumulating zeaxanthin and canthaxanthin, the resulting transformants produced their 2-hydroxylated and 2,2'-dihydroxylated products which were structurally novel or rare xanthophylls, as determined by their nuclear magnetic resonance and high-performance liquid chromatography/photodiode array detector/atmospheric pressure chemical ionization mass spectrometry spectral data. The new carotenoid produced was suggested to have a strong inhibitory effect on lipid peroxidation.
Volume 71(8)
Pages 4286-96
Published 2005-8-1
DOI 10.1128/AEM.71.8.4286-4296.2005
PII 71/8/4286
PMID 16085816
PMC PMC1183362
MeSH Amino Acid Sequence Animals Bacterial Proteins / genetics Bacterial Proteins / metabolism Biotechnology / methods Carotenoids / biosynthesis* Carotenoids / chemistry Carotenoids / genetics Caulobacteraceae / enzymology Caulobacteraceae / genetics* Caulobacteraceae / metabolism Escherichia coli / enzymology Escherichia coli / genetics Escherichia coli / metabolism Humans Mice Mixed Function Oxygenases / genetics* Mixed Function Oxygenases / metabolism Molecular Sequence Data Multigene Family* Xanthophylls / biosynthesis*
IF 4.016
DNA material pAC-Cantha (RDB19488) pAC-Asta (RDB19489) pUCBre-W (RDB19515) pUCBre-O11 (RDB19516)