Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction, SunMoonUniversity, #100, Kalsan-ri, Tangjeong-myeon, Asan-si, Chungnam 336-708, Republic of Korea.
Enzyme Microb Technol. 2013 Apr 10;52(4-5):234-40. doi: 10.1016/j.enzmictec.2013.02.004. Epub 2013 Feb 16.
A sterol glucosyltransferase-encoded gene was isolated from Salinispora tropica CNB-440, a marine, sediment-dwelling, Gram positive bacterium that produces the potent anticancer compound, salinosporamide A. The full-length gene consists of 1284 nucleotides and encodes 427 amino acids with a calculated mass of 45.65kDa. The gene was then cloned and heterologously expressed in Escherichia coli BL21(DE3). The amino acid sequence shares 39% similarity with the glycosyltransferase from Withania somnifera, which belongs to glycosyltransferase family 1. Enzyme reactions were carried out with the various free sterols (acceptor) and NDP-sugars (donor). The purified protein only showed activity for glucosylation of β-sitosterol with UDP-D-glucose and TDP-D-glucose donors, and optimal activity at pH 7.5 and 37°C. Among these two donors, UDP-D-glucose was preferred.
从海洋沉积物来源的革兰氏阳性细菌盐孢菌（Salinispora tropica） CNB-440 中分离到一种甾醇葡糖基转移酶编码基因，该菌能产生强效抗癌化合物——柳氮磺胺吡啶 A。全长基因由 1284 个核苷酸组成，编码 427 个氨基酸，理论分子量为 45.65kDa。该基因随后被克隆并在大肠杆菌 BL21(DE3)中异源表达。该氨基酸序列与属于糖基转移酶家族 1 的睡茄中的糖基转移酶有 39%的相似性。用各种游离甾醇（受体）和 NDP-糖（供体）进行酶反应。纯化的蛋白仅对 β-谷甾醇的 UDP-D-葡萄糖和 TDP-D-葡萄糖供体具有葡糖基化活性，在 pH7.5 和 37°C 时具有最佳活性。在这两种供体中，UDP-D-葡萄糖是首选。
