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Applied and Environmental Microbiology, March 2007, p. 1736-1741, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02425-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of a Sterol {Delta}7 Reductase Gene Involved in Desmosterol Biosynthesis in Mortierella alpina 1S-4{triangledown} ,{dagger}

Shuo Zhang, Eiji Sakuradani, and Sakayu Shimizu*

Division of Applied Life of Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan

Received 16 October 2006/ Accepted 1 January 2007

Molecular cloning of the gene encoding sterol {Delta}7 reductase from the filamentous fungus Mortierella alpina 1S-4, which accumulates cholesta-5,24-dienol (desmosterol) as the main sterol, revealed that the open reading frame of this gene, designated Mo{Delta}7SR, consists of 1,404 bp and codes for 468 amino acids with a molecular weight of 53,965. The predicted amino acid sequence of Mo{Delta}7SR showed highest homology of 51% with that of sterol {Delta}7 reductase (EC 1.3.1.21) from Xenopus laevis (African clawed frog). Heterologous expression of the Mo{Delta}7SR gene in yeast Saccharomyces cerevisiae revealed that Mo{Delta}7SR converts ergosta-5,7-dienol to ergosta-5-enol (campesterol) by the activity of {Delta}7 reductase. In addition, with gene silencing of Mo{Delta}7SR gene by RNA interference, the transformant accumulated cholesta-5,7,24-trienol up to 10% of the total sterols with a decrease in desmosterol. Cholesta-5,7,24-trienol is not detected in the control strain. This indicates that Mo{Delta}7SR is involved in desmosterol biosynthesis in M. alpina 1S-4. This study is the first report on characterization of sterol {Delta}7 reductase from a microorganism.

* Corresponding author. Mailing address: Division of Applied Life of Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan. Phone: 81757536114. Fax: 81757536128. E-mail: sim{at}kais.kyoto-u.ac.jp.

{triangledown} Published ahead of print on 12 January 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, March 2007, p. 1736-1741, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02425-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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