The K428 residue from Thermus thermophilus SG0.5JP17-16 laccase plays the substantial role in substrate binding and oxidation
The K428 residue from Thermus thermophilus SG0.5JP17-16 laccase plays the substantial role in substrate binding and oxidation
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来自嗜热栖热菌 SG0.5JP17-16 漆酶的 K428 残基在底物结合和氧化中发挥重要作用
DOI：
10.1080/07391102.2020.1729864
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发表时间：
2020-02
影响因子：
4.4
通讯作者：
Yang Xiaorong
中科院分区：
文献类型：
--
作者：
Zhu Yanyun;Zhan Jiangbo;Zhang Yi;Lin Ying;Yang Xiaorong
摘要漆酶是一种多铜氧化酶，能催化多种有机和无机化合物的氧化，同时还能将分子氧还原为水，被认为是一种绿色催化剂。以往的研究表明，在MCOs中，底物结合口袋的结构与底物特异性密切相关。口袋结构环上的氨基酸参与底物的识别并影响酶的催化性质。嗜热栖热菌SG 0. 5 JP 17 -16漆酶（lacTT）的Lys 428残基位于口袋环上，为了探讨K428残基的作用，获得催化活性增强的突变体，采用定点突变的方法将其突变为亮氨酸、谷氨酸、精氨酸和蛋氨酸。结构数据显示，K428残基通过空间位阻和与底物的相互作用识别底物的构象。动力学数据表明，甲硫氨酸取代K428导致突变体具有增强的催化活性。K428 M突变体对合成染料的脱色效果优于野生型。本研究为寻找催化活性增强的突变体提供了策略。作者：Ramaswamy H. Sarma
Abstract Laccases are multi-copper oxidases (MCOs) that catalyze the oxidation of various organic and inorganic compounds with concomitant reduction of dioxygen to water, and are considered to be green catalysts. Previous studies revealed that in MCOs, the structure of substrate-binding pocket was closely related to the substrate specificity. The amino acids on the pocket-constructing loops were involved in the identification of substrate and affected the catalytic properties of the enzyme. In the Thermus thermophilus SG0.5JP17-16 laccase (lacTT), the Lys428 residue is on pocket-constructing loops, to explore the role of the K428 residue and obtain mutants with enhanced catalytic activity, it was mutated to leucine, glutamic acid, arginine and methionine by site-directed mutagenesis. Structural data revealed that the K428 residue identified the conformation of the substrate by the steric hindrance and interactions with the substrate. Kinetic data indicated that the replacement of K428 by methionine resulted in a mutant with enhanced catalytic activity. The K428M mutant could decolorize the synthetic dyes more efficiently than the wild type enzyme. Altogether, this study provided a strategy to find the mutant with the enhanced catalytic activity. Communicated by Ramaswamy H. Sarma
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嗜热栖热菌 SG0.5JP17-16 漆酶的四个第二球残基通过氢键网络调节催化作用
影响因子：
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作者：
Liu Huiping;Zhu Yanyun;Yang Xiaorong;Lin Ying
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影响因子：
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通讯作者：
FINAZZIAGRO, A
DOI：
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发表时间：
2005-08-09
影响因子：
11.1
作者：
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通讯作者：
Kramer, KJ
影响因子：
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作者：
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DOI：
10.1107/s2053230x1600426x
发表时间：
2016-04
期刊：
Acta crystallographica. Section F, Structural biology communications
影响因子：
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作者：
Liu Z;Xie T;Zhong Q;Wang G
通讯作者：
Wang G