Structure and Engineering of L-Arabinitol 4-Dehydrogenase from Neurospora crassa
Structure and Engineering of L-Arabinitol 4-Dehydrogenase from Neurospora crassa
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DOI：
10.1016/j.jmb.2010.07.033
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发表时间：
2010-09-10
影响因子：
5.6
通讯作者：
Nair, Satish K.
中科院分区：
文献类型：
--
作者：
Bae, Brian;Sullivan, Ryan P.;Nair, Satish K.
L-阿拉伯糖醇4-脱氢酶（LAD）催化L-阿拉伯糖醇转化为L-木酮糖，同时伴有NAD（+）还原。它是利用真菌L-阿拉伯糖分解代谢途径将L-阿拉伯糖转化为燃料和化学品的重组生物体开发中的必需酶。在这里，我们报告的晶体结构的LAD从丝状真菌粗糙脉孢菌在2.6埃分辨率。此外，我们还创建了N的一些定点变体。crassa LAD，其能够利用NADP（+）作为辅因子，产生具有几乎完全转换的辅因子特异性的LAD的第一个实例。这项工作代表了任何LAD的第一个结构数据，并为理解有关该酶的底物特异性和辅因子特异性的现有文献提供了分子基础。具有改变的辅因子特异性的工程化LAD突变体在工业生物技术中的应用是有用的。(C)2010爱思唯尔有限公司保留所有权利。
L-Arabinitol 4-dehydrogenase (LAD) catalyzes the conversion of L-arabinitol into L-xylulose with concomitant NAD(+) reduction. It is an essential enzyme in the development of recombinant organisms that convert L-arabinose into fuels and chemicals using the fungal L-arabinose catabolic pathway. Here we report the crystal structure of LAD from the filamentous fungus Neurospora crassa at 2.6 angstrom resolution. In addition, we created a number of site-directed variants of N. crassa LAD that are capable of utilizing NADP(+) as cofactor, yielding the first example of LAD with an almost completely switched cofactor specificity. This work represents the first structural data on any LAD and provides a molecular basis for understanding the existing literature on the substrate specificity and cofactor specificity of this enzyme. The engineered LAD mutants with altered cofactor specificity should be useful for applications in industrial biotechnology. (C) 2010 Elsevier Ltd. All rights reserved.