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

Authors
Tomiyasu, K Yato, K Yasuda, M Tonozuka, T Ibuka, A Sakai, H
Citation
K. Tomiyasu et al., Cloning and nucleotide sequence of the pullulanase gene of Thermus thermophilus HB8 and production of the enzyme in Escherichia coli, BIOS BIOT B, 65(9), 2001, pp. 2090-2094
Citations number
19
Categorie Soggetti
Agricultural Chemistry","Biochemistry & Biophysics
Journal title
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
ISSN journal
09168451 → ACNP
Volume
65
Issue
9
Year of publication
2001
Pages
2090 - 2094
Database
ISI
SICI code
0916-8451(200109)65:9<2090:CANSOT>2.0.ZU;2-U
Abstract
A 3.4-kb SphI fragment carrying the pullulanase gene of Thermus thermophilu s HB8 was cloned. Based on the nucleotide sequence of it and the flanking r egion analyzed by direct sequencing of the inverse PCR product, an expressi on vector was constructed. The E. coli cells harboring the plasmid produced an about 80-kDa protein having pullulanase activity, the optimum temperatu re of which was 70 degreesC.