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Applied and Environmental Microbiology, August 2005, p. 4286-4296, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4286-4296.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Elucidation of a Carotenoid Biosynthesis Gene Cluster Encoding a Novel Enzyme, 2,2'-ß-Hydroxylase, from Brevundimonas sp. Strain SD212 and Combinatorial Biosynthesis of New or Rare Xanthophylls

Yasuhiro Nishida,1 Kyoko Adachi,2 Hiroaki Kasai,2 Yoshikazu Shizuri,2 Kazutoshi Shindo,3 Akiyoshi Sawabe,1 Sadao Komemushi,1 Wataru Miki,4 and Norihiko Misawa2*

School of Agriculture, Kinki University, Nakamachi, Nara,1 Marine Biotechnology Institute, Heita, Kamaishi, Iwate,2 Japan Women's University, Mejirodai, Bunkyo-ku, Tokyo,3 Suntory Limited, Shimamoto-cho, Mishima-gun, Japan4

Received 19 December 2004/ Accepted 3 March 2005

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'-ß-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.

* Corresponding author. Mailing address: Marine Biotechnology Institute, 3-75-1, Heita, Kamaishi, Iwate 026-0001, Japan. Phone: 81 193 26 6581. Fax: 81 193 26 6584. E-mail: norihiko.misawa{at}mbio.jp.

Applied and Environmental Microbiology, August 2005, p. 4286-4296, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4286-4296.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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