Cloning of a gene cluster for dextrin utilization from Thermoactinomyces vulgaris R-47 and characterization of the cyclodextrin-binding protein

To investigate the physiological role of an a-amylase, TVA II from Thermoactinomyces vulgaris R-47, which hydrolyze cyclodextrins and pullulan, a region located upstream of the TVA II gene was cloned and sequenced. Five open reading frames, designated as ORF-1 to -5, were found in a fragment of abou...

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Bibliographic Details
Published in:Journal of Applied Glycoscience 2002/04/01, Vol.49(2), pp.107-114
Main Authors: Yopi (Tokyo Univ. of Agriculture and Technology, Fuchu (Japan). Faculty of Agriculture), Tonozuka, T, Sakai, H, Sakano, Y
Format: Article
Language:English
Subjects:
ALPHA AMYLASE
DEXTRINS
GENES
MOLECULAR CLONING
PROTEINS
THERMOACTINOMYCES
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Summary:To investigate the physiological role of an a-amylase, TVA II from Thermoactinomyces vulgaris R-47, which hydrolyze cyclodextrins and pullulan, a region located upstream of the TVA II gene was cloned and sequenced. Five open reading frames, designated as ORF-1 to -5, were found in a fragment of about 5 kbp. Three of these genes, ORF-1, -2 and -3, were homologous to the genes which encode proteins related to the sugar metabolic systems, such a maltose system of Escherichia coli and cyclodextrin metabolic system of Klebsiella oxytoca. An expression vector for the ORF-3 protein, which was similar to maltose-binding protein from E. coli (MalE) and a cyclodextrin-binding protein from K. oxytoca (CymE), was constructed, and the expressed ORF-3 protein was purified. The ORF-3 protein was a cyclodextrin-binding protein because of having a higher binding ability for cyclodextrins than for maltose.
ISSN:1344-7882
1880-7291
DOI:10.5458/jag.49.107