来自斋藤曲霉的1,2-α-D-甘露糖苷酶基因msdS的分子克隆、核苷酸序列及其在酵母细胞中的表达
Molecular cloning and nucleotide sequence of the 1,2-alpha-D-mannosidase gene, msdS, from Aspergillus saitoi and expression of the gene in yeast cells.
作者信息
Inoue T, Yoshida T, Ichishima E
机构信息
Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan.
出版信息
Biochim Biophys Acta. 1995 Dec 6;1253(2):141-5. doi: 10.1016/0167-4838(95)00195-6.
A full-length cDNA encoding 1,2-alpha-D-mannosidase (EC 3.2.1.113) from Aspergillus saitoi was cloned. Analysis of the 1718 bp nucleotide sequence of the cDNA revealed a single open reading frame with 1539 nucleotides of 1,2-alpha-D-mannosidase gene, msdS. The predicted amino-acid sequence of 1,2-alpha-D-mannosidase consists of 513 residues with a molecular mass of 55,767 and is 70%, 26% and 35% identity with those of Penicillium citrinum 1,2-alpha-D-mannosidase, yeast alpha-mannosidase, and mouse alpha-mannosidase. The cDNA of the msdS gene has been cloned and expressed in yeast cells. To identify the activity of expression product methyl-2-O-alpha-mannopyranosyl-alpha-mannopyranoside (Man alpha 1-->2Man-OMe) was used as a substrate at pH 5.0.
克隆了来自斋藤曲霉的编码1,2-α-D-甘露糖苷酶（EC 3.2.1.113）的全长cDNA。对该cDNA的1718 bp核苷酸序列分析显示，存在一个1539个核苷酸的1,2-α-D-甘露糖苷酶基因msdS的单一开放阅读框。预测的1,2-α-D-甘露糖苷酶氨基酸序列由513个残基组成，分子量为55767，与桔青霉1,2-α-D-甘露糖苷酶、酵母α-甘露糖苷酶和小鼠α-甘露糖苷酶的氨基酸序列分别具有70%、26%和35%的同一性。msdS基因的cDNA已被克隆并在酵母细胞中表达。为鉴定表达产物的活性，在pH 5.0条件下，以甲基-2-O-α-甘露吡喃糖基-α-甘露吡喃糖苷（Manα1→2Man-OMe）作为底物。
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