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Galdino AS, Silva RN, Lottermann MT, et al. Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae. Enzyme Res. 2011;2011:157294doi: 10.4061/2011/157294.
Galdino, A. S., Silva, R. N., Lottermann, M. T., Alvares, A. C., de Moraes, L. M., Torres, F. A., de Freitas, S. M., & Ulhoa, C. J. (2011). Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae. Enzyme research, 2011157294. https://doi.org/10.4061/2011/157294
Galdino, Alexsandro Sobreira, et al. "Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae." Enzyme research vol. 2011 (2011): 157294. doi: https://doi.org/10.4061/2011/157294
Galdino AS, Silva RN, Lottermann MT, Alvares AC, de Moraes LM, Torres FA, de Freitas SM, Ulhoa CJ. Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae. Enzyme Res. 2011 Mar 30;2011:157294. doi: 10.4061/2011/157294. PMID: 21490699; PMCID: PMC3068306.
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Enzyme Res. 2011 Mar 30;2011:157294. doi: 10.4061/2011/157294.

Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae.

Enzyme research

Alexsandro Sobreira Galdino, Roberto Nascimento Silva, Muriele Taborda Lottermann, Alice Cunha Morales Alvares, Lídia Maria Pepe de Moraes, Fernando Araripe Gonçalves Torres, Sonia Maria de Freitas, Cirano José Ulhoa

Affiliations

  1. Laboratório de Biotecnologia, Universidade Federal de São João del-Rei, 35501-296 Divinópolis, MG, Brazil.

PMID: 21490699 PMCID: PMC3068306 DOI: 10.4061/2011/157294

Abstract

An extracellular alpha-amylase (Amy1) whose gene from Cryptococcus flavus was previously expressed in Saccharomyces cerevisiae was purified to homogeneity (67 kDa) by ion-exchange and molecular exclusion chromatography. The enzyme was activated by NH(4) (+) and inhibited by Cu(+2) and Hg(+2). Significant biochemical and structural discrepancies between wild-type and recombinant α-amylase with respect to K(m) values, enzyme specificity, and secondary structure content were found. Far-UV CD spectra analysis at pH 7.0 revealed the high thermal stability of both proteins and the difference in folding pattern of Amy1 compared with wild-type amylase from C. flavus, which reflected in decrease (10-fold) of enzymatic activity of recombinant protein. Despite the differences, the highest activity of Amy1 towards soluble starch, amylopectin, and amylase, in contrast with the lowest activity of Amy1(w), points to this protein as being of paramount biotechnological importance with many applications ranging from food industry to the production of biofuels.

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