广西科学2017，Vol.24Issue(1)：100-105,6.DOI:10.13656/j.cnki.gxkx.20170117.001
嗜碱芽孢杆菌耐热耐碱内切木聚糖酶基因的密码子优化及在毕赤酵母中的表达
The Codon Optimization of Thermostable and Alkali-resisting Endo-xylanase from Bacillus halodurans S7 and Expression in Pichia pastoris
摘要
Abstract
[Objective]Studying the extreme heat and alkali xylanase can satisfy the harsh environment with high temperature and alkaline in papermaking industry.[Methods]A xylanase gene derived from an alkalophilic Bacillus S7 (xyn10A) is codon optimized, then cloned into pHBM905BDM vector, and transformed into Pichia pastoris strain GS115.[Results]The similarity between the optimized xylanase gene (xyn10A) and the original sequence is 76.70%.Then the gene is cloned into pHBM905BDM vector and successfully transformed into P.pastoris GS115.The transformations are screened on the RBB-xylan plate, then in the shake flask fermentation a high expression strain is obtained.The highest activity is 495 U/mL at the 10th day of the shake fermentation.In addition, glycosylation analysis shows that the enzyme is glycosylated in P.pastoris.[Conclusion]The optimized extreme heat and alkali resistance xylanase gene are successfully expressed in P.pastoris.关键词
嗜碱芽孢杆菌内切木聚糖酶/基因优化/表达/毕赤酵母Key words
thermostable and alkali-resisting xylanase/codon optimization/expression/Pichia pastoris分类
生物科学引用本文复制引用
严闯,尹李峰,周玉玲,张桂敏..嗜碱芽孢杆菌耐热耐碱内切木聚糖酶基因的密码子优化及在毕赤酵母中的表达[J].广西科学,2017,24(1):100-105,6.基金项目
国家"863"高科技项目(2012AA022203C)资助. （2012AA022203C）
