Rational affinity purification of native Streptomyces family 10 xylanase
| Autor: | Shigeyasu Ito, Atsushi Kuno, Satoshi Kaneko, Yasuyuki Kawabata, Ryuichiro Suzuki, Isao Kusakabe, Tsunemi Hasegawa |
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| Rok vydání: | 2004 |
| Předmět: |
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Western Bioengineering Biology Disaccharides medicine.disease_cause Applied Microbiology and Biotechnology Streptomyces Chromatography Affinity Sepharose Affinity chromatography medicine Binding site Escherichia coli Streptomycetaceae General Medicine biology.organism_classification Recombinant Proteins Xylosidases Biochemistry Xylanase Electrophoresis Polyacrylamide Gel Xylans Carbohydrate-binding module Protein Binding Biotechnology |
| Zdroj: | Journal of Biotechnology. 110:137-142 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/j.jbiotec.2004.01.014 |
| Popis: | Xylanase SoXyn10A from Streptomyces olivaceoviridis E-86 comprises a family 10 catalytic module linked to a family 13 carbohydrate-binding module (SoCBM13). The SoCBM13 has a beta-trefoil structure, with binding sites in each subdomain (alpha, beta and gamma). Subdomain alpha, but not subdomains beta and gamma, binds tightly to lactose. It was, therefore, thought that immobilized lactose could be used for the affinity purification of SoXyn10A. Lactosyl-Sepharose was prepared and tested as an affinity matrix. SoXyn10A produced from the cloned xyn10A gene by Escherichia coli, and native SoXyn10A in culture supernatants from S. olivaceoviridis, were purified to homogeneity in a single step by affinity chromatography using this matrix. This simple purification of SoXyn10A makes the enzyme an attractive candidate for applications requiring xylanase. The CBM also has the potential for use as an affinity tag for the purification of other proteins. |
| Databáze: | OpenAIRE |
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