Abstract
A novel dextransucrase gene, DSRN, was obtained by ultrasoft X-ray treatment of the DSRB742 gene. The DSRN gene was further mutated via site-directed mutagenesis producing four mutants: DSRN1 (F196S), DSRN2 (Y346N), DSRN3 (K395T) and DSRN4 (P980T). Dextransucrases derived from DSRB742 and its mutants were expressed in E. coli and affinity-purified using dextran to give 80% purity. They had specific activities of 0.6–17 U/mg with Km values of 18–88 mM. DSRB742 had the lowest (0.02 s−1 · mM−1) and DSRN1 had the highest (0.13 s−1 · mM−1) Kcat/Km values. DSRN3 had the highest enzymatic transglycosylation efficiency with maltose (63% of theoretical), gentiobiose (39%), or salicine (40%).
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Acknowledgements
This work was mostly supported by International Cooperative Research Program, Korea Science and Engineering Foundation (Grant M60402010002-05A0201-00210), and partially by Post-doc supporting program of Korean Research Foundation Grant (KRF-2005-216-C00055) and CNU Specialization Grant by Chonnam National University for students.
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Nam, S.H., Ko, E.A., Jang, S.S. et al. Maximization of dextransucrase activity expressed in E. coli by mutation and its functional characterization. Biotechnol Lett 30, 135–143 (2008). https://doi.org/10.1007/s10529-007-9498-z
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DOI: https://doi.org/10.1007/s10529-007-9498-z