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Journal of Bacteriology, September 2002, p. 5151-5157, Vol. 184, No. 18
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.18.5151-5157.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

barS1, a Gene for Biosynthesis of a {gamma}-Butyrolactone Autoregulator, a Microbial Signaling Molecule Eliciting Antibiotic Production in Streptomyces Species

Noriyasu Shikura, Junji Yamamura, and Takuya Nihira*

Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan

Received 19 April 2002/ Accepted 19 June 2002

From Streptomyces virginiae, in which production of streptogramin antibiotic virginiamycin M1 and S is tightly regulated by a low-molecular-weight Streptomyces hormone called virginiae butanolide (VB), which is a member of the {gamma}-butyrolactone autoregulators, the hormone biosynthetic gene (barS1) was cloned and characterized by heterologous expression in Escherichia coli and by gene disruption in S. virginiae. The barS1 gene (a 774-bp open reading frame encoding a 257-amino-acid protein [Mr, 27,095]) is situated in the 10-kb regulator island surrounding the VB-specific receptor gene, barA. The deduced BarS1 protein is weakly homologous to ß-ketoacyl-acyl carrier protein/coenzyme A reductase and belongs to the superfamily of short-chain alcohol dehydrogenase. The function of the BarS1 protein in VB biosynthesis was confirmed by BarS1-dependent in vitro conversion of 6-dehydro-VB-A to VB-A, the last catalytic step in VB biosynthesis. Of the four possible enantiomeric products from racemic 6-dehydro-VB-A as a substrate, only the natural enantiomer of (2R,3R,6S)-VB-A was produced by the purified recombinant BarS1 (rBarS1), indicating that rBarS1 is the stereospecific reductase recognizing (3R)-isomer as a substrate and reducing it stereospecifically to the (6S) product. In the {Delta}barS1 mutant created by homologous recombination, the production of VB as well as the production of virginiamycin was lost. The production of virginiamycin by the {Delta}barS1 mutant was fully recovered by the external addition of VB to the culture, which indicates that the barS1 gene is essential in the biosynthesis of the autoregulator VBs in S. virginiae and that the failure of virginiamycin production was a result of the loss of VB production.

* Corresponding author. Permanent address: The International Center for Biotechnology, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan. Phone: 81-6-6879-7433. Fax: 81-6-6879-7432. E-mail: nihira{at}icb.osaka-u.ac.jp.

Journal of Bacteriology, September 2002, p. 5151-5157, Vol. 184, No. 18
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.18.5151-5157.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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