来自[具体来源未明确]的重组莨菪碱-6β-羟化酶的分离、表达及生化特性研究——调节东莨菪碱的产量
Isolation, expression and biochemical characterization of recombinant hyoscyamine-6β-hydroxylase from - tuning the scopolamine production.
作者信息
Fischer Conrad, Kwon Moonhyuk, Ro Dae-Kun, van Belkum Marco J, Vederas John C
机构信息
University of Alberta , Department of Chemistry , Edmonton , AB T6G 2G2 , Canada . Email:
University of Calgary , Biological Sciences , Calgary , AB T2N 1N4 , Canada.
出版信息
Medchemcomm. 2018 May 2;9(5):888-892. doi: 10.1039/c8md00090e. eCollection 2018 May 1.
Abstract
Hyoscyamine-6β-hydroxylase (H6H, EC 1.14.11.11) is a plant enzyme that catalyses the last two steps in the biosynthesis of the anticholinergic drug scopolamine, the hydroxylation of hyoscyamine to 6β-hydroxyhyoscyamine (anisodamine) and subsequent oxidative ring-closure to the 6,7-β-epoxide. A gene homologue was isolated from the plant () and recombinantly cloned into , expressed and purified using an effective SUMO-fusion procedure. Enzymatic activity is approximately 40-fold higher for the first reaction step and the substrate affinity is comparable to other characterized H6H homologues ( ∼ 60 μM). Truncation of an H6H enzyme flexible N-terminal region yields an active and stable yet more compact enzyme version.
摘要
莨菪碱-6β-羟化酶（H6H，EC 1.14.11.11）是一种植物酶，催化抗胆碱能药物东莨菪碱生物合成的最后两步，即将莨菪碱羟基化为6β-羟基莨菪碱（山莨菪碱），随后氧化闭环生成6,7-β-环氧化物。从植物（此处原文缺失植物名称）中分离出一个基因同源物，并通过有效的SUMO融合程序进行重组克隆、表达和纯化。第一个反应步骤的酶活性大约高40倍，底物亲和力与其他已表征的H6H同源物相当（约60μM）。截短H6H酶的柔性N端区域可产生一种活性和稳定性更高但结构更紧凑的酶变体。
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