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Expression, Characterization and Synergistic Interactions of Myxobacter Sp. AL-1 Cel9 and Cel48 Glycosyl Hydrolases
Instituto de Investigación en Biología Experimental (IIBE), Facultad de Química, Universidad de Guanajuato, P.O. Box 187, Guanajuato, Gto. 36050, Mexico
Instituto de Ciencias de la Salud, Universidad Autónoma del Estado de Hidalgo, Abasolo 600, Pachuca, Hgo 42000, Mexico
* Author to whom correspondence should be addressed.
Received: 2 December 2007; in revised form: 24 January 2008 / Accepted: 1 February 2008 / Published: 29 February 2008
Abstract: The soil microorganism Myxobacter Sp. AL-1 regulates in a differential manner the production of five extracellular cellulases during its life cycle. The nucleotide sequence of a cel9-cel48 cluster from the genome of this microorganism was recently obtained. Cel48 was expressed in Escherichia coli to generate a His6-Cel48 protein and the biochemical properties of the pure protein were determined. Cel48 was more efficient in degrading acid-swollen avicel (ASC) than carboxymethylcellulose (CMC). On the other hand, cel9 was expressed in Bacillus subtilis from an IPTG-inducible promoter. Zymogram analysis showed that after IPTG-induction, Cel9 existed in both the cell fraction and the culture medium of B. subtilis and the secreted protein was purified to homogeneity by FPLC-ionic exchange chromatography. The exocellobiohydrolase Cel48 showed a synergism of 1.68 times with the endocellulase Cel9 during ASC degradation using an 8.1- fold excess of Cel48 over Cel9. Western blot analysis revealed that both proteins were synthesized and secreted to the culture medium of Myxobacter Sp. AL-1. These results show that the cel9-cel48 cluster encodes functional endo- and exo-acting cellulases that allows Myobacter Sp. AL-1 to hydrolyse cellulose.
Keywords: Cellulose; Cel9; Cel48 cellulases; Synergism; Myxobacter Sp. AL-1.
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Ramírez-Ramírez, N.; Romero-García, E.R.; Calderón, V.C.; Avitia, C.I.; Téllez-Valencia, A.; Pedraza-Reyes, M. Expression, Characterization and Synergistic Interactions of Myxobacter Sp. AL-1 Cel9 and Cel48 Glycosyl Hydrolases. Int. J. Mol. Sci. 2008, 9, 247-257.
Ramírez-Ramírez N, Romero-García ER, Calderón VC, Avitia CI, Téllez-Valencia A, Pedraza-Reyes M. Expression, Characterization and Synergistic Interactions of Myxobacter Sp. AL-1 Cel9 and Cel48 Glycosyl Hydrolases. International Journal of Molecular Sciences. 2008; 9(3):247-257.
Ramírez-Ramírez, Norma; Romero-García, Eliel R.; Calderón, Vianney C.; Avitia, Claudia I.; Téllez-Valencia, Alfredo; Pedraza-Reyes, Mario. 2008. "Expression, Characterization and Synergistic Interactions of Myxobacter Sp. AL-1 Cel9 and Cel48 Glycosyl Hydrolases." Int. J. Mol. Sci. 9, no. 3: 247-257.