Original Article

The Journal of Antibiotics (2006) 59, 471–479; doi:10.1038/ja.2006.66

Geosmin Biosynthesis in Streptomyces avermitilis. Molecular Cloning, Expression, and Mechanistic Study of the Germacradienol/Geosmin Synthase

David E Cane1, Xiaofei He1, Seiji Kobayashi3, Satoshi O macrmura2 and Haruo Ikeda3

  1. 1Department of Chemistry, Brown University, Box H, Providence, Rhode Island 02912-9108 USA
  2. 2The Kitasato Institute, 9-1, Shirokane 5-chome Minato-ku, Tokyo 108-8642, Japan
  3. 3Kitasato Institute for Life Sciences, Kitasato University, 1-15-1, Kitasato, Sagamihara, Kanagawa 228-8555, Japan

Correspondence: D. E. Cane, Department of Chemistry, Brown University, Box H, Providence, Rhode Island 02912-9108 USA. E-mail: David_Cane@brown.edu

Received 30 May 2006; Accepted 3 August 2006.

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Abstract

Geosmin (1) is responsible for the characteristic odor of moist soil. The Gram-positive soil bacterium Streptomyces avermitilis produces geosmin (1) as well as its precursor germacradienol (3). The S. avermitilis gene SAV2163 (geoA) is extremely similar to the S. coelicolor A3(2) SCO6073 gene that encodes a germacradienol/geosmin synthase. S. avermitilis mutants with a deleted geoA were unable to produce either germacradienol (3) or geosmin (1). Biosynthesis of both compounds was restored by introducing an intact geoA gene into the mutants. Incubation of recombinant GeoA, encoded by the SAV2163 gene of S. avermitilis, with farnesyl diphosphate (2) in the presence of Mg2+ gave a mixture of (4S,7R)-germacra-1(10)E,5E-diene-11-ol (3) (66%), (7S)-germacrene D (4) (24%), geosmin (1) (8%), and a hydrocarbon, tentatively assigned the structure of octalin 5 (2%). Incubation of this germacradienol/geosmin synthase with [1,1-2H2]FPP (2a) gave geosmin-d1 (1a), as predicted. When recombinant GeoA from either S. avermitilis or S. coelicolor A3(2) was incubated with nerolidyl diphosphate (8), only the acyclic elimination products beta-farnesene (10), (Z)-alpha-farnesene (11), and (E)-alpha-farnesene (12) were formed, thereby ruling out nerolidyl diphosphate as an intermediate in the conversion of farnesyl diphosphate to geosmin, germacradienol, and germacrene D.

Keywords:

sesquiterpene, biosynthesis, geosmin, Streptomyces avermitilis, Streptomyces coelicolor

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