Abstract
The mutant strain PN-120 of Cellulomonas flavigena produces a ß-glucosidase that is 10-fold more active than the corresponding enzyme isolated from the parental strain. These enzymes were partially purified through Q Sepharose and Bio-Gel filtration. A single protein band was detected on polyacrylamide–gel electrophoresis/zymogram using 4-methylumbelliferyl-β-D-glucoside. On sodium dodecyl sulfate–PAGE, the enzyme displayed three protein bands, suggesting that in C. flavigena the enzyme is oligomeric with a molecular mass of 210 kDa. On purification, the specific activity of ß-glucosidase isolated from PN-120 was increased 16-fold and showed three times more affinity for cellobiose than the enzyme of the parental strain; nevertheless, the optimum pH and temperature were similar for both enzymes. The kinetic parameters suggested that the increase in the activity of the enzyme, from the mutant strain, was caused by a mutation that affects the catalytic site of the enzyme. The partial amino-acid sequence of the isolated enzyme confirmed that it is a β-glucosidase because of its homology with other β-glucosidases produced by cellulolytic bacteria and fungi.
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Acknowledgments
This work was supported by CONACyT-México(45678-Z). A. Barrera-Islas received a scholarship from CONACyT- México. The investigators thank O. Pérez-Avalos and M. Mercado-Morales for technical assistance.
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Barrera-Islas, G., Ramos-Valdivia, A., Salgado, L. et al. Characterization of a β-Glucosidase Produced by a High-Specific Growth-Rate Mutant of Cellulomonas flavigena . Curr Microbiol 54, 266–270 (2007). https://doi.org/10.1007/s00284-006-0105-7
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DOI: https://doi.org/10.1007/s00284-006-0105-7