Functional identification of rub52 gene involved in the biosynthesis of rubradirin.
An open reading frame, rub52, has been identified as a gene encoding thymidine diphospho-glucose 2,3-dehydratase by sequence analysis of the rubradirin biosynthetic gene cluster of Streptomyces achromogenes var. nibradiris NRRL3061. The gene codes for a protein consisting of 458 amino acids with calculated molecular mass of 50862 Da. The gene was amplified and heterologously expressed in Escherichia coli as a soluble His-tagged fusion protein. C-2 deoxygenation functionality of thymidine diphospho-4-keto-6-deoxyglucose was assigned to the rub52 gene product from in vitro enzyme assay.
Maharjan
Jyoti
J
Institute of Biomolecule Reconstruction, Sun Moon University, #100, Kalsan-ri, Tangjeong-myeon, Asan-si, Chungnam 336-708, South Korea.
Liou
Kwangkyoung
K
Lee
Hei Chan
HC
Kim
Chun-Gyu
CG
Lee
Jung-Joon
JJ
Yoo
Jin-Cheol
JC
Sohng
Jae Kyung
JK
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D006027:Glycosides / Q000235:genetics / Q000378:metabolism; D006836:Hydro-Lyases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009285:Naphthoquinones / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*
D006027:Glycosides; D009285:Naphthoquinones; D011993:Recombinant Fusion Proteins; C024513:rubranitrose; C002857:rubradirin; D006836:Hydro-Lyases; C022771:dTDPglucose 4,6-dehydratase