Three Molecular Modification Strategies to Improve the Thermostability of Xylanase XynA from Streptomyces rameus L2001.
Three Molecular Modification Strategies to Improve the Thermostability of Xylanase XynA from Streptomyces rameus L2001.
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提高拉米链霉菌 L2001 木聚糖酶 XynA 热稳定性的三种分子修饰策略
DOI：
10.3390/foods12040879
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发表时间：
2023-02-18
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影响因子：
--
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中科院分区：
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糖苷水解酶家族11 （GH11）木聚糖酶是生产功能性低聚糖的首选候选酶。然而，天然GH11木聚糖酶的低热稳定性限制了它们的工业应用。在本研究中，我们研究了以下三种策略来改变链霉菌L2001突变木聚糖酶XynA的热稳定性，以减少表面熵、分子内二硫键的构建和分子环化。利用分子模拟分析了XynA突变体的热稳定性变化。除分子环化外，所有突变体的热稳定性和催化效率均优于XynA。高熵氨基酸替代突变体Q24A和K104A在65℃保存30 min后，残活由18.70%增加到41.23%以上。以山毛榉木聚糖为底物时，Q24A和K143A的催化效率分别为129.99和92.26 mL/s/mg，而XynA的催化效率为62.97 mL/s/mg。与野生型XynA相比，Val3和Thr30之间形成二硫键的突变酶的t1/260°C和催化效率分别提高了13.33倍和1.80倍。XynA突变体具有较高的热稳定性和水解活性，将有助于酶促生产功能性低聚木糖。
Glycoside hydrolase family 11 (GH11) xylanases are the preferred candidates for the production of functional oligosaccharides. However, the low thermostability of natural GH11 xylanases limits their industrial applications. In this study, we investigated the following three strategies to modify the thermostability of xylanase XynA from Streptomyces rameus L2001 mutation to reduce surface entropy, intramolecular disulfide bond construction, and molecular cyclization. Changes in the thermostability of XynA mutants were analyzed using molecular simulations. All mutants showed improved thermostability and catalytic efficiency compared with XynA, except for molecular cyclization. The residual activities of high-entropy amino acid-replacement mutants Q24A and K104A increased from 18.70% to more than 41.23% when kept at 65 °C for 30 min. The catalytic efficiencies of Q24A and K143A increased to 129.99 and 92.26 mL/s/mg, respectively, compared with XynA (62.97 mL/s/mg) when using beechwood xylan as the substrate. The mutant enzyme with disulfide bonds formed between Val3 and Thr30 increased the t1/260 °C by 13.33-fold and the catalytic efficiency by 1.80-fold compared with the wild-type XynA. The high thermostabilities and hydrolytic activities of XynA mutants will be useful for enzymatic production of functional xylo-oligosaccharides.
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影响因子：
64.8
作者：
Glenwright AJ;Pothula KR;Bhamidimarri SP;Chorev DS;Baslé A;Firbank SJ;Zheng H;Robinson CV;Winterhalter M;Kleinekathöfer U;Bolam DN;van den Berg B
通讯作者：
van den Berg B
通过 N 端延伸工程和 C 端定点诱变改善嗜热踝节菌 F1208 木聚糖酶的特殊水解特性
DOI：
10.1016/j.ijbiomac.2016.12.050
发表时间：
2017-03-01
影响因子：
8.2
作者：
Li, Qin;Sun, Baoguo;Li, Xiuting
通讯作者：
Li, Xiuting
影响因子：
3.7
作者：
He, Jun;Tang, Feng;Yu, Feng
通讯作者：
Yu, Feng
影响因子：
8
作者：
Grant, Barry J.;Skjaerven, Lars;Yao, Xin-Qiu
通讯作者：
Yao, Xin-Qiu
影响因子：
3.9
作者：
Boonyaputthikul, Harit;Muhammad, Auwal;Sutthibutpong, Thana
通讯作者：
Sutthibutpong, Thana