Alteromonas macleodii木聚糖酶XynZT-3序列分析及表达研究  

Sequence analysis and expression of xylanase XynZT-3 from Alteromonas macleodii

作　　者：石嘉宁 徐佳 孔梦圆 王宁宁 田艳杰 崔彩霞 周晨妍 SHI Jianing;XU Jia;KONG Mengyuan;WANG Ningning;TIAN Yanjie;CUI Caixia;ZHOU Chenyan(Xinxiang Medical University,Synthetic Biology Engineering Laboratory of Henan Province,School of Life Science and Technology,Xinxiang,Henan 453003)

机构地区：[1]新乡医学院生命科学技术学院河南省合成生物学工程实验室,河南新乡453003

出　　处：《工业微生物》2022年第2期48-55,共8页Industrial Microbiology

基　　金：河南省教育厅重点研究课题(21A180024);河南省科技厅科技攻关项目(212102210652)。

摘　　要：为了拓宽海洋微生物来源木聚糖酶在食品工业中的应用,利用生物信息学对实验室保藏的麦氏交替单胞菌木聚糖酶XynZT-3基因进行序列分析,将xynZT-3基因连入pPIC9K表达载体,经SalⅠ线性化后,电击转化毕赤酵母GS115,并对构建的重组菌进行诱导表达条件优化。结果显示:该序列全长2970 bp,编码989个氨基酸,无内含子和信号肽,预测分子量为107.74 kDa,属于GH10家族。重组菌P.pastoris GS115/xynZT-3经单因素优化及响应面分析后,在16℃、甲醇浓度1.50%、种龄28 h、pH 3.7,培养6 d的最优摇瓶条件下,重组酶活力为4.212 U/mL。本研究将为该酶的进一步研究奠定基础。In order to broaden the application of marine microbial xylanase in food industry,the sequence of xylanase XynZT-3 gene from Alteromonas macleodii was analyzed by bioinformatics.The target gene was inserted into the Pichia pastoris expression vector pPIC9 K.After linearized by Sal Ⅰ,the recombinant vector was transformed into Pichia pastoris GS115 by electroporation,and the induced expression conditions were optimized.The results showed that the full length of the sequence was 2 970 bp,encoding 989 amino acids,no introns and signal peptides,and the predicted molecular weight was 107.74 kDa,which belonged to the GH10 family.After single factor optimization and response surface analysis,the optimum shake flask conditions for the recombinant strain P.pastoris GS115/xynZT-3 were as follows:temperature 16 ℃,methanol concentration 1.50%,inoculation age 28 h,pH 3.7,and induction time 6 days.Under the optimized conditions,the recombinant enzyme activity was 4.212 U/mL.This research would lay a foundation for the further research of this enzyme.

关 键 词：麦氏交替单胞菌 木聚糖酶 序列分析 单因素优化 响应面分析 

分 类 号：Q78[生物学—分子生物学]