Cloning and nucleotide sequence of the pullulanase gene of Thermus thermophilus HB8 and production of the enzyme in Escherichia coli

被引：9

作者：

Tomiyasu, K ^{[1
]}

Yato, K ^{[1
]}

Yasuda, M ^{[1
]}

Tonozuka, T ^{[1
]}

Ibuka, A ^{[1
]}

Sakai, H ^{[1
]}

机构：

[1] Univ Shizuoka, Dept Food & Nutrit Sci, Grad Sch Nutrit & Environm Sci, Shizuoka 4228526, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2001年 / 65卷 / 09期

关键词：

pullulanase; Thermus thermophilus; debranching enzyme; gene cloning; inverse PCR;

D O I：

10.1271/bbb.65.2090

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A 3.4-kb SphI fragment carrying the pullulanase gene of Thermus thermophilus HB8 was cloned. Based on the nucleotide sequence of it and the flanking region analyzed by direct sequencing of the inverse PCR product, an expression vector was constructed. The E. coli cells harboring the plasmid produced an about 80-kDa protein having pullulanase activity, the optimum temperature of which was 70 degreesC.

引用

页码：2090 / 2094

页数：5

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