Functional characterization of ketoreductase (rubN6) and aminotransferase (rubN4) genes in the gene cluster of Streptomyces achromogenes var. rubradiris.
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.
Lamichhane
Janardan
J
Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction, IBR, SunMoon University, Tangjeonmyun, 336-708, Asan-si, Chung-nam, Republic of Korea.
Liou
Kwangkyoung
K
Lee
Hei Chan
HC
Kim
Chun-Gyu
CG
Sohng
Jae Kyung
JK
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002237:Carbohydrate Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family / Q000235:genetics*; D009285:Naphthoquinones / Q000737:chemistry / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D000637:Transaminases / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D009285:Naphthoquinones; C002857:rubradirin; D010088:Oxidoreductases; D002237:Carbohydrate Dehydrogenases; C514865:thymidine diphosphate glucose 4-ketoreductase, Streptomyces achromogenes; D000637:Transaminases; C514866:thymidine diphosphate glucose 3-aminotransferase, Streptomyces achromogenes