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| | ONLINE | ISSN | : | 1347-6947 | | PRINT | ISSN | : | 0916-8451 |
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| | Bioscience, Biotechnology, and Biochemistry |
| Vol. 66 (2002) , No. 12 pp.2594-2599 |
| [PDF (543K)] [References]  |
 | Cloning of the Maltose Phosphorylase Gene from Bacillus sp. Strain RK-1 and Efficient Production of the Cloned Gene and the Trehalose Phosphorylase Gene from…
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| | | 1) Showa Sangyo Co., Ltd. |
| (Received June 7, 2002)
(Accepted August 4, 2002)
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| | The maltose phosphorylase (MPase) gene of Bacillus sp. strain RK-1 was cloned by PCR with oligonucleotide primers designed on the basis of a partial N-terminal amino acid sequence of the purified enzyme. The MPase gene consisted of 2,655 bp encoding a theoretical protein with a Mr of 88,460, and had no secretion signal sequence, although most of the MPase activity was detected in the culture supernatant of RK-1. This cloned MPase gene and the trehalose phosphorylase (TPase) gene from Bacillus stearothermophilus SK-1 were efficiently expressed intracellularly under the control of the Bacillus amyloliquefaciens α-amylase promoter in Bacillus subtilis. The production yields were estimated to be more than 2 g of enzyme per liter of medium, about 250 times the production of the original strains, in a simple shake flask. About 60% of maltose was converted into trehalose by the simultaneous action of both enzymes produced in B. subtilis.
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| To cite this article: | | Yasushi INOUE, Nozomu YASUTAKE, Yoshie OSHIMA, Yoshie YAMAMOTO, Tetsuji TOMITA, Shinsuke MIYOSHI and Tsuneya YATAKE, “Cloning of the Maltose Phosphorylase Gene from Bacillus sp. Strain RK-1 and Efficient Production of the Cloned Gene and the Trehalose Phosphorylase Gene from…”, Biosci. Biotechnol. Biochem., Vol. 66, 2594-2599 (2002) . | |
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| doi:10.1271/bbb.66.2594 | | JOI JST.JSTAGE/bbb/66.2594 |
| Copyright (c) 2003 by Japan Society for Bioscience, Biotechnology, and Agrochemistry |
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