Functional characterization of ketoreductase (rubN6) and aminotransferase (rubN4) genes in the gene cluster of Streptomyces achromogenes var. rubradiris.

Biotechnol Lett

Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction, IBR, SunMoon University, Tangjeonmyun, 336-708, Asan-si, Chung-nam, Republic of Korea.

Published: April 2006

ORF's for rubN6 and rubN4 have been annotated as thymidine diphosphate glucose 4-ketoreductase and thymidine diphosphate glucose 3-aminotransferase by sequence analysis of the rubradirin biosynthetic gene cluster cloned from Streptomyces achromogenes var. rubradiris NRRL 3061. Both ORFs were heterologously expressed in Escherichia coli as His-tagged fusion proteins. The functionalities of TDP-glucose 4-ketoreductase and TDP-glucose 3-aminotransferase were verified by in vitro enzyme assay, and a biosynthetic pathway for TDP-D: -rubranitrose is proposed.

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http://dx.doi.org/10.1007/s10529-006-0013-8DOI Listing

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