摘要
以水生栖热菌FL-03菌株的基因组DNA为模板,PCR扩增编码嗜热麦芽糖转葡萄糖基酶的基因(MalQ)片段,克隆至pET-28a(+)原核表达载体,转化E.coli BL21(DE3),IPTG诱导表达,对表达产物进行SDS-PAGE和Western blotting分析鉴定。重组蛋白经70℃钝化离心除杂蛋白后,经SDS-PAGE检测,可见单一条带。纯化酶液以10%麦芽糖为底物,经TLC法分析表明该酶具有转葡萄糖基的活性,比活力提高近5倍,且活力回收率达70%,具有重要的工业应用前景。
MalQ gene was amplified by PCR with Thermus aquaticus FL-03 genome DNA, the amplified MalQ gene was cloned into prokaryotic expression vector pET-28a（＋） and expressed in BL21 （DE3） under induction of IPTG. The expressed product was identified with SDS-PAGE and Western blotting. The amylomaltase was purified by inactivation at 70 ℃ and centrifugalization. The glucanothansferase activity of the purified amylomaltase was identified via TLC method with 10% maltose as substrate. The specific activity is improved 5 folds and the residual activity is 70% after purification. The amylomaltase has great potential in application of various industries.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2008年第12期389-392,共4页
Food Science
基金
吉林省“十一五”重大科技攻关项目(20065006)
