Tian Yong-Sheng, Xu Jing, Chen Lei, Fu Xiao-Yan, Peng Ri-He, Yao Quan-Hong
Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute of Shanghai Academy of Agricultural Sciences, Shanghai 201106, P.R. China.
Shanghai Ruifeng Agricultural Science and Technology Co., Ltd, Shanghai 201106, P.R. China.
J Microbiol Biotechnol. 2017 Oct 28;27(10):1783-1789. doi: 10.4014/jmb.1705.05026.
Thermostability is an important property of xylanase because high temperature is required for its applications, such as wood pulp bleaching, baking, and animal feedstuff processing. In this study, from , a moderately thermophilic gram-negative bacterium, was modified via site-directed mutagenesis (based on its 3D structure) to obtain thermostable xylanase, and the properties of this enzyme were analyzed. Results revealed that the half-life of xylanase at 65°C increased from 10 to 50 min after a disulfide bridge was introduced between the α-helix and its adjacent β-sheet at S98 and N145. Further mutation at the side of A153E named XynB-CE in the C-terminal of this α-helix enhanced the half-life of xylanase for 60 min at 65°C. Therefore, the mutant may be utilized for industrial applications.
热稳定性是木聚糖酶的一项重要特性,因为其应用(如木浆漂白、烘焙和动物饲料加工)需要高温。在本研究中,来自嗜温革兰氏阴性菌的木聚糖酶通过定点诱变(基于其三维结构)进行改造,以获得热稳定木聚糖酶,并对该酶的特性进行了分析。结果表明,在S98和N145的α-螺旋与其相邻β-折叠之间引入二硫键后,木聚糖酶在65°C的半衰期从10分钟增加到50分钟。在该α-螺旋C端A153E位点进一步突变得到的XynB-CE,使木聚糖酶在65°C的半衰期延长至60分钟。因此,该突变体可用于工业应用。