芝田硫化叶菌新型α-淀粉酶基因在大肠杆菌的克隆和表达  

CLONING AND EXPRESSION OF THE GENE ENCODING NOVEL α-AMYLASE FROM SULFOLOBUS SHIBATAE IN E.COLI

作　　者：刘莉[1] 陈炜[1] 金城[2] 

机构地区：[1]中国科学院微生物研究所酶学研究室,北京100080 [2]中国科学院微生物研究所微生物资源国家重点实验室,北京100080

出　　处：《微生物学报》2000年第3期323-326,共4页Acta Microbiologica Sinica

基　　金：微生物资源国家重点实验室课题! ( 981 0 2 4 );北京市自然科学基金!课题 ( 5982 0 1 0 )

摘　　要：A novel α amylase gene was amplified from Sulfolobus shibatae by using PCR technique. The amplified 1.7kb DNA fragment was inserted into an expression vector pBV220 to yield the recombinant plasmid pSBAM. The novel α amylase gene in pSBAM was expressed in E. coli . The production of the novel α amylase activity reached over 8 units/100mL of the culture. The molecular weight of this enzyme was about 61kD by SDS PAGE. The expressed novel α amylase protein in E.coli DH5α accounted for about 20% of the total protein in the recombinant cell. The cooperative action of the novel α amylase and the maltooligosyltrehalose synthase from Sulfolobus shibatae was investigated and trehalose was detected by using HPLC analysis when using amylose and partial starch hydrolysates as substrates.A novel α amylase gene was amplified from Sulfolobus shibatae by using PCR technique. The amplified 1.7kb DNA fragment was inserted into an expression vector pBV220 to yield the recombinant plasmid pSBAM. The novel α amylase gene in pSBAM was expressed in E. coli . The production of the novel α amylase activity reached over 8 units/100mL of the culture. The molecular weight of this enzyme was about 61kD by SDS PAGE. The expressed novel α amylase protein in E.coli DH5α accounted for about 20% of the total protein in the recombinant cell. The cooperative action of the novel α amylase and the maltooligosyltrehalose synthase from Sulfolobus shibatae was investigated and trehalose was detected by using HPLC analysis when using amylose and partial starch hydrolysates as substrates.

关 键 词：Α-淀粉酶 基因克隆 基因表达 海藻糖 酶法合成 

分 类 号：Q785[生物学—分子生物学]