Enhancing H2O2 resistance of an esterase from Pyrobaculum calidifontis by structure-guided engineering of the substrate binding site
Enhancing H2O2 resistance of an esterase from Pyrobaculum calidifontis by structure-guided engineering of the substrate binding site
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通过底物结合位点的结构引导工程增强 Pyrobaculum calidifontis 酯酶的 H2O2 抗性
DOI：
10.1007/s00253-017-8299-0
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发表时间：
2017-05
影响因子：
5
通讯作者：
Wang YH
中科院分区：
文献类型：
--
作者：
Zhou Pengfei;Yang Bo;Lan Dongming;Wang Xuping;Wang Yonghua;Popowicz Grzegorz Maria;Wang YH
在工业化学领域，绿色技术正受到越来越多的关注，其中酶促反应能够替代危险且对环境不友好的化学工艺。过氧羧酸的原位酶促合成是一种很有吸引力的替代方法。
Green technologies are attracting increasing attention in industrial chemistry where enzymatic reactions can replace dangerous and environmentally unfriendly chemical processes. In situ enzymatic synthesis of peroxycarboxylic acid is an attractive alternative for several industrial applications although concentrated H2O2can denature the biocatalyst, limiting its usefulness. Herein, we report the structure-guided engineering of the Pyrobaculum calidifontis esterase (PestE) substrate binding site to increase its stability and perhydrolysis activity. The L89R/L40A PestE mutant showed better tolerance toward concentrated H2O2compared with wild-type PestE,and retained over 72% of its initial activity after 24-h incubation with 2 M H2O2. Surprisingly, the half-life (t1/2, 80 °C) of PestE increased from 28 to 54 h. The kcat/Kmvalues of the mutant increased 21- and 3.4-fold toward pentanoic acid and H2O2,respectively. This work shows how protein engineering can be used to enhance the H2O2resistance and catalytic efficiency of an enzyme.
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影响因子：
16.6
作者：
Bernhardt, P;Halt, K;Kazlauskas, RJ
通讯作者：
Kazlauskas, RJ
影响因子：
2.1
作者：
BJORKLING, F;FRYKMAN, H;KIRK, O
通讯作者：
KIRK, O
影响因子：
2.1
作者：
M. Moreira;T. B. Bitencourt;M. G. Nascimento
通讯作者：
M. Moreira;T. B. Bitencourt;M. G. Nascimento
影响因子：
3.9
作者：
Barry J. Ryan;C. Ó’Fágáin
通讯作者：
Barry J. Ryan;C. Ó’Fágáin
影响因子：
3.5
作者：
Ullrich, Rene;Dolge, Christoph;Hofrichter, Martin
通讯作者：
Hofrichter, Martin