摘要
用ＰＣＲ 方法从芝田硫化叶菌中扩增了编码一种新酶，即麦芽寡糖基海藻糖合酶（ ＭＴＳａｓｅ） 的基因，扩增的２－２ｋｂ ＤＮＡ 插入到原核表达载体ｐＢＶ２２０ 中，构建成重组质粒ｐＳＢＧＴ１ 。ｐＳＢＧＴ１ 中ＭＴＳａｓｅ 基因在大肠杆菌中得到表达。ＳＤＳＰＡＧＥ 分析表达产物ＭＴＳａｓｅ蛋白的分子量约为７４ｋＤａ ，同核苷酸序列测定所推导的值相符。表达产物占细胞总蛋白约４－４ ％ 。ｐＳＢＧＴ１ 产生的重组酶作用于淀粉部分水解物，使ＤＥ 值降低，得到非还原糖或低还原糖。
kb DNA fragment encoding a novel enzyme, maltooligosy1 trehalose synthase (MTSase) was amplified from\% Sulfolobus shibatae\% by using PCR technique. The amplified 2.2kb DNA fragment was inserted into an expression vector, pBV220, to yield the recombinant plasmid pSBGT1. MTSase gene in pBSGT1 was expressed in \%E.coli\%. The molecular weight of expressed MTSase detected by SDS\|PAGE was about 74kD, which is conformed with that deduced from nucleotide sequence. The expressed MTSase protein accounted for about 4.4% of the total cell protein. The MTSase from transformants containing pBSGT1 is capable of decreasing DE value, forming non\|reducing or less\|reducing saccharides when allowed to act on reducing partial starch hydrolysates.
出处
《微生物学报》
CAS
CSCD
北大核心
2000年第1期57-61,共5页
Acta Microbiologica Sinica
基金
北京市自然科学基金
微生物资源国家重点实验室课题
关键词
MTSASE
海藻糖
酶法
大肠杆菌
基因克隆
Maltooligosyl trehalose synthase, Sulfolobus shibatae, Gene cloning and expression, E.coil
