Sianidis Giorgos, Wohlert Sven-Eric, Pozidis Charalambos, Karamanou Spyridoula, Luzhetskyy Andriy, Vente Andreas, Economou Anastassios
Institute of Molecular Biology and Biotechnology, FORTH and Department of Biology, University of Crete, P.O. Box 1527, Iraklio-Crete 71110, Greece.
J Biotechnol. 2006 Sep 18;125(3):425-33. doi: 10.1016/j.jbiotec.2006.03.035. Epub 2006 May 19.
We have cloned the gene that encodes a novel glucosyl transferase (AraGT) involved in rhamnosylation of the polyketide antibiotic Aranciamycin in Streptomyces echinatus. AraGT comprises two domains characteristic of bacterial glycosyltranferases. AraGT was synthesized in E. coli as a decahistidinyl-tagged polypeptide. Purified AraGT is dimeric, displays a T(mapp) of 30 degrees C and can glycosylate the aglycone of an Aranciamycin derivative as shown by liquid chromatography and mass spectrometry. The availability of functional AraGT will allow the generation Aranciamycin-based combinatorial libraries.
我们已经克隆了编码一种新型葡糖基转移酶（AraGT）的基因，该酶参与棘孢链霉菌中聚酮类抗生素阿霉素的鼠李糖基化作用。AraGT包含细菌糖基转移酶特有的两个结构域。AraGT在大肠杆菌中作为带有十聚组氨酸标签的多肽合成。纯化后的AraGT是二聚体，其表观熔解温度为30℃，并且如液相色谱和质谱所示，它能够使阿霉素衍生物的糖苷配基糖基化。功能性AraGT的获得将有助于构建基于阿霉素的组合文库。
