Cloning, Sequencing, and Expression of the Gene Encoding an Intracellular β-D-Xylosidase from Streptomyces thermoviolaceus OPC-520
| العنوان: | Cloning, Sequencing, and Expression of the Gene Encoding an Intracellular β-D-Xylosidase from Streptomyces thermoviolaceus OPC-520 |
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| المؤلفون: | Mitsuo Kosaka, Chiaki Takada, Hiroshi Tsujibo, Katsushiro Miyamoto, Akihiko Tsuji, Yoshihiko Inamori |
| المصدر: | Bioscience, Biotechnology, and Biochemistry. 65:1824-1831 |
| بيانات النشر: | Oxford University Press (OUP), 2001. |
| سنة النشر: | 2001 |
| مصطلحات موضوعية: | Molecular Sequence Data, Biology, Molecular cloning, medicine.disease_cause, Applied Microbiology and Biotechnology, Biochemistry, Analytical Chemistry, Gene product, Escherichia coli, medicine, Amino Acid Sequence, Cloning, Molecular, Molecular Biology, Peptide sequence, chemistry.chemical_classification, Base Sequence, Hydrolysis, Organic Chemistry, Nucleic acid sequence, General Medicine, Chromosomes, Bacterial, Molecular biology, Recombinant Proteins, Streptomyces, Culture Media, Amino acid, Open reading frame, Xylosidases, chemistry, Genes, Bacterial, Electrophoresis, Polyacrylamide Gel, Plasmids, Biotechnology, Streptomyces thermoviolaceus |
| الوصف: | The intracellular beta-xylosidase was induced when Streptomyces thermoviolaceus OPC-520 was grown at 50 degrees C in a minimal medium containing xylan or xylooligosaccharides. The 82-kDa protein with beta-xylosidase activity was partially purified and its N-terminal amino acid sequence was analyzed. The gene encoding the enzyme was cloned, sequenced, and expressed in Escherichia coli. The bxlA gene consists of a 2,100-bp open reading frame encoding 770 amino acids. The deduced amino acid sequence of the bxlA gene product had significant similarity with beta-xylosidases classified into family 3 of glycosyl hydrolases. The bxlA gene was expressed in E. coli, and the recombinant protein was purified to homogeneity. The enzyme was a monomer with a molecular mass of 82 kDa. The purified enzyme showed hydrolytic activity towards only p-nitrophenyl-beta-D-xylopyranoside among the synthetic glycosides tested. Thin-layer chromatography analysis showed that the enzyme is an exo-type enzyme that hydrolyze xylooligosaccharides, but had no activity toward xylan. High activity against pNPX occurred in the pH range 6.0-7.0 and temperature range 40-50 degrees C. |
| تدمد: | 1347-6947 0916-8451 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::41b5800074773e99aca3a4f01414f7ff https://doi.org/10.1271/bbb.65.1824 |
| رقم الأكسشن: | edsair.doi.dedup.....41b5800074773e99aca3a4f01414f7ff |
| قاعدة البيانات: | OpenAIRE |
PDF full text from Publisher | تدمد: | 13476947 09168451 |
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