摘要
Bacillus amyoliquefaciens DL-3纤维素酶具有热稳定高比活多功能的酶学特性,本文根据该酶的氨基酸序列合成了其编码基因(cel),构建了pPIC9K-cel表达载体,并用P.pastoris进行了表达。工程菌株三角瓶发酵酶活性达0.50 U/mL,酶解滤纸的产物为低聚糖,表明人工合成的热稳定高比活纤维素酶基因在P.pastoris中可以正常表达、加工及分泌,重组酶的分子量由天然酶的53 kDa增加至68 kDa,糖基化严重。
A complete gene was synthesized according to the amino acid sequence of Bacillus amyoliquefaciens DL-3 cellulase and expressed extracellularly in Pichia pastoris. The CMC activity of the recombinant P. pastoris strain reached 0.5 U/mL in Erlenmeyer flask, and the hydrolytic products of paper by the recombined cellulase were oligosaccharides. The molecular mass of the recombinant enzyme was found to be about 68 kDa by SDS PAGE, which was much higher than 53 kDa of the nature one. Glycosylation was presumed to be the cause of low activity.
出处
《工业微生物》
CAS
CSCD
2010年第5期27-31,共5页
Industrial Microbiology
基金
北京市自然科学基金项目(5083025)
国家高技术研究发展计划(863计划
2007AA091601-2)
中国农业科学院科技经费项目
关键词
纤维素酶
解淀粉芽胞杆菌
毕赤酵母
糖基化
cellulase
Bacillus amyoliquefaciens
Pichia pastoris
glycosilation
