摘要
采用简并PCR和TAIL-PCR技术,从来源于天山冰川土壤的类芽孢杆菌Paenibacillussp.A1菌株基因组中扩增出β-甘露聚糖酶基因编码序列。序列分析表明,该基因全长960bp,编码一个新的糖苷水解酶第5家族的β-甘露聚糖酶。将该基因克隆到原核表达载pET-22b(+)中,经过IPTG诱导表达,酶的胞外表达量达0.62U/mL。酶学特性分析表明其最适pH为6.5,最适温度为55℃,属中性β-甘露聚糖酶,具有较好的底物适应性,适宜用于鱼类等水产动物养殖饲料行业中。
A β-mannanase encoding gene was isolated from strain of Paenibacillus sp.A1 by degenerate PCR and TAIL-PCR.Sequence analysis shows that the gene of 960 bp length encodes a novel β-mannanase,which belongs to glycosyl hydrolase family 5.The gene was ligated into the prokaryotic expression vector pET-22b（＋）,then expressed with the enzyme activity of 0.62 U/mL after induction by IPTG.Enzymatic characteristics analysis showed that the optimum pH for the enzyme was 6.5 and the optimum temperature was 55℃.It is a neutral β-mannanase,which has better substrate adaptability. It is suitable to be taken as feed additives for fish and other aquaculture industry.
出处
《中国农业科技导报》
CAS
CSCD
2009年第5期60-65,共6页
Journal of Agricultural Science and Technology
基金
农业微生物菌种资源整理整合及共享项目(2005DKA21201)
国家863计划项目(2006AA02Z213)资助
