Lamichhane Janardan, Liou Kwangkyoung, Lee Hei Chan, Kim Chun-Gyu, Sohng Jae Kyung
Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction, IBR, SunMoon University, Tangjeonmyun, 336-708, Asan-si, Chung-nam, Republic of Korea.
Biotechnol Lett. 2006 Apr;28(8):545-53. doi: 10.1007/s10529-006-0013-8.
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.
通过对从产色链霉菌变种红色产色链霉菌NRRL 3061克隆的rubradirin生物合成基因簇进行序列分析，rubN6和rubN4的开放阅读框被注释为胸苷二磷酸葡萄糖4-酮还原酶和胸苷二磷酸葡萄糖3-氨基转移酶。这两个开放阅读框均在大肠杆菌中作为His标签融合蛋白进行了异源表达。通过体外酶分析验证了TDP-葡萄糖4-酮还原酶和TDP-葡萄糖3-氨基转移酶的功能，并提出了TDP-D：-红曲糖的生物合成途径。
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