Biochemical Characterization of GacI, a Bifunctional Glycosyltransferase–Phosphatase Enzyme Involved in Acarbose Biosynthesis in Streptomyces glaucescens GLA.O,Biochemistry

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Biochemical Characterization of GacI, a Bifunctional Glycosyltransferase–Phosphatase Enzyme Involved in Acarbose Biosynthesis in Streptomyces glaucescens GLA.O
Biochemistry ( IF 2.9 ) Pub Date : 2022-10-26 , DOI: 10.1021/acs.biochem.2c00473
Takeshi Tsunoda ₁ , Shumpei Asamizu ₁ , Taifo Mahmud ₁

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Acarbose, a pseudotetrasaccharide produced by several strains of Actinoplanes and Streptomyces, is an α-glucosidase inhibitor clinically used to control type II diabetes. Bioinformatic analysis of the biosynthetic gene clusters of acarbose in Actinoplanes sp. SE50/110 (the acb cluster) and Streptomyces glaucescens GLA.O (the gac cluster) revealed their distinct genetic organizations and presumably biosynthetic pathways. However, to date, only the acarbose pathway in the SE50/110 strain has been extensively studied. Here, we report that GacI, one of the proteins that appear to be different between the two pathways, is a bifunctional glycosyltransferase family 5 (GT5)–phosphatase (PP) enzyme that functions at two different steps in acarbose biosynthesis in S. glaucescens GLA.O. In the acb pathway, the GT and the PP reactions are performed by two different enzymes. Truncated GacI proteins having only the GT or the PP domain showed comparable catalytic activity with the full-length GacI, indicating that domain separation does not significantly affect their respective catalytic activity. GacI, which is widely distributed in many Streptomyces, represents the first example of naturally occurring GT5-PP bifunctional enzymes biochemically characterized.

中文翻译：

GacI 的生化表征，一种双功能糖基转移酶-磷酸酶，参与青绿链霉菌 GLA.O 中阿卡波糖的生物合成

阿卡波糖是由几种游动放线菌和链霉菌菌株产生的假四糖，是一种α-葡萄糖苷酶抑制剂，临床上用于控制II型糖尿病。游动动动菌阿卡波糖生物合成基因簇的生物信息学分析。SE50/110（acb簇）和Streptomyces glaucescens GLA.O（gac簇）揭示了它们独特的遗传组织和可能的生物合成途径。然而，迄今为止，仅对 SE50/110 菌株中的阿卡波糖途径进行了广泛研究。在这里，我们报道 GacI 是两种途径之间似乎不同的蛋白质之一，它是一种双功能糖基转移酶家族 5 (GT5) - 磷酸酶 (PP) 酶，在青芒草 GLA 中阿卡波糖生物合成的两个不同步骤中发挥作用.O. 在acb途径中，GT 和 PP 反应由两种不同的酶进行。仅具有 GT 或 PP 结构域的截短 GacI 蛋白表现出与全长 GacI 相当的催化活性，表明结构域分离不会显着影响它们各自的催化活性。GacI 广泛分布于许多链霉菌中，代表了天然存在的 GT5-PP 双功能酶生化特征的第一个例子。

更新日期：2022-10-26

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