The structure of a novel glucuronoyl esterase from Myceliophthora thermophila gives new insights into its role as a potential biocatalyst
The structure of a novel glucuronoyl esterase from Myceliophthora thermophila gives new insights into its role as a potential biocatalyst
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DOI：
10.1107/s0907444912042400
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发表时间：
2013-01-01
影响因子：
2.2
通讯作者：
Chrysina, Evangelia D.
中科院分区：
文献类型：
--
作者：
Charavgi, Maria-Despoina;Dimarogona, Maria;Chrysina, Evangelia D.
开发高效生物催化剂的需求日益增长是其广泛工业应用的结果。典型的困难，在他们的开发过程中遇到的各种过程是相互关联的因素，如温度，pH值，产品抑制剂等，以消除这些，研究已经指向新的酶，将符合所需的标准鉴定。为此，最近发现的葡萄糖醛酸酯酶（GE）是碳水化合物酯酶（CE）家族中一个神秘的家族。来自嗜热毁丝霉（Myceliophthora thermophila）（同义词嗜热孢子丝菌（Sporotrichum thermophile））（CE 15家族的成员）及其S213 A突变体的嗜热StGE 2酯酶的结构分别在1.55和1.9埃分辨率下测定。在2.35埃分辨率下测定了与底物类似物4-O-甲基-β-D-吡喃葡萄糖醛酸甲酯复合的S213 A突变体的第一晶体结构。所有的三维蛋白质结构都具有α/β-水解酶折叠，其具有三层α β α-夹心结构和Rossmann拓扑结构，并且每个不对称单元包含一个分子。这些是第一个晶体结构的嗜热GE无论是在一个unliganded形式和绑定到一个底物类似物，从而解开组织的催化三联体残基和他们的邻居内衬的活性位点。所获得的知识为驱动葡萄糖醛酸酯水解的关键结构元件提供了新的见解。
The increasing demand for the development of efficient biocatalysts is a consequence of their broad industrial applications. Typical difficulties that are encountered during their exploitation in a variety of processes are interconnected with factors such as temperature, pH, product inhibitors etc. To eliminate these, research has been directed towards the identification of new enzymes that would comply with the required standards. To this end, the recently discovered glucuronoyl esterases (GEs) are an enigmatic family within the carbohydrate esterase (CE) family. Structures of the thermophilic StGE2 esterase from Myceliophthora thermophila (synonym Sporotrichum thermophile), a member of the CE15 family, and its S213A mutant were determined at 1.55 and 1.9 angstrom resolution, respectively. The first crystal structure of the S213A mutant in complex with a substrate analogue, methyl 4-O-methyl-beta-D-glucopyranuronate, was determined at 2.35 angstrom resolution. All of the three-dimensional protein structures have an alpha/beta-hydrolase fold with a three-layer alpha beta alpha-sandwich architecture and a Rossmann topology and comprise one molecule per asymmetric unit. These are the first crystal structures of a thermophilic GE both in an unliganded form and bound to a substrate analogue, thus unravelling the organization of the catalytic triad residues and their neighbours lining the active site. The knowledge derived offers novel insights into the key structural elements that drive the hydrolysis of glucuronic acid esters.