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Identification of α-amylase by random and specific mutagenesis of Texcoconibacillus texcoconensis 13CCT strain isolated from extreme alkaline–saline soil of the former Lake Texcoco (Mexico)

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Abstract

The alkaline α-amylase produced by Texcoconibacillus texcoconensis 13CCT strain was identified by random mutagenesis and confirmed by directed mutagenesis. A transposon mutagenesis approach was taken to identify the gene responsible for the degradation of starch in T. texcoconensis 13CCT strain. The deduced amino acids of the amy gene had a 99 % similarity with those of Bacillus selenitireducens MLS10 and 97 % with those of Paenibacillus curdlanolyticus YK9. The enzyme showed a maximum activity of 131.1 U/mL at 37 °C and pH 9.5 to 10.5. In situ activity of the enzyme determined by polyacrylamide gel electrophoresis showed only one band with amylolytic activity. This is the first report of a bacterium isolated from the extreme alkaline–saline soil of the former Lake Texcoco (Mexico) with amylolytic activity in alkaline conditions while its potential as a source of amylases for the industry is discussed.

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Acknowledgments

This work was funded by the ‘Instituto de Ciencia y Tecnología del Distrito Federal (Mexico)’ project ICyTDF/295/2009. J.M. B.-L., Y.E. N.-N, S. G.-A, and Z. H.-M. received grant-aided support from ‘Consejo Nacional de Ciencia y Tecnología’ (CONACYT, Mexico). Escherichia coli BW20767 carrying plasposon pRL27 and E. coli DH5α/λpir were kindly provided by Dr. William W. Metcalf (University of Illinois at Urbana–Champaign, USA). Escherichia coli S17-1 λpir carrying the plasmid pKNOCK-Cm was provided by Dr. Mikhail F. Alexeyev (University of South Alabama, Mobile, AL, USA).

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Authors and Affiliations

  1. Laboratory of Soil Ecology, Department of Biotechnology and Bioengineering, ABACUS, CINVESTAV, Av. I.P.N. 2508, C.P. 07360, Mexico, D.F., Mexico

    Juan Manuel Bello-López, Yendi E. Navarro-Noya, Selene Gómez-Acata & Luc Dendooven

  2. Laboratory of Industrial Microbiology, Department of Microbiology, Escuela Nacional de Ciencias Biológicas, IPN, Prolongación de Carpio y Plan de Ayala S/N, Col. Santo Tomás, C. P. 11340, Mexico, D.F., Mexico

    Zahuiti Hernández-Montañez

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  1. Juan Manuel Bello-López
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  2. Yendi E. Navarro-Noya
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  3. Selene Gómez-Acata
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  4. Zahuiti Hernández-Montañez
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  5. Luc Dendooven
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Correspondence to Luc Dendooven.

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Bello-López, J.M., Navarro-Noya, Y.E., Gómez-Acata, S. et al. Identification of α-amylase by random and specific mutagenesis of Texcoconibacillus texcoconensis 13CCT strain isolated from extreme alkaline–saline soil of the former Lake Texcoco (Mexico). Folia Microbiol 59, 235–240 (2014). https://doi.org/10.1007/s12223-013-0289-8

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  • DOI: https://doi.org/10.1007/s12223-013-0289-8

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