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Studies on a thermostable α-amylase from the thermophilic fungus Scytalidium thermophilum

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Abstract

An α-amylase produced by Scytalidium thermophilum was purified using DEAE-cellulose and CM-cellulose ion exchange chromatography and Sepharose 6B gel filtration. The purified protein migrated as a single band in 6% PAGE and 7% SDS-PAGE. The estimated molecular mass was 36 kDa (SDS-PAGE) and 49 kDa (Sepharose 6B). Optima of pH and temperature were 6.0 and 60°C, respectively. In the absence of substrate the purified α-amylase was stable for 1 h at 50°C and had a half-life of 12 min at 60°C, but was fully stable in the presence of starch. The enzyme was not activated by several metal ions tested, including Ca2+ (up to 10 mM), but HgCl2 and CuCl2 inhibited its activity. The α-amylase produced by S. thermophilum preferentially hydrolyzed starch, and to a lesser extent amylopectin, maltose, amylose and glycogen in that order. The products of starch hydrolysis (up to 6 h of reaction) analyzed by thin layer chromatography, showed oligosaccharides such as maltotrioses, maltotetraoses and maltopentaoses. Maltose and traces of glucose were formed only after 3 h of reaction. These results confirm the character of the enzyme studied to be an α-amylase (1,4-α-glucan glucanohydrolase).

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Acknowledgements

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho de Desenvolvimento Científico e Tecnológico (CNPq). H.F.T., J.A.J. and M.L.T.M.P. are Research Fellows of CNPq. A.C.M.M.A. was the recipient of a CAPES Fellowship. This work was part of a Master Dissertation submitted by A.C.M.M.A. to the Departamento de Biologia, FFCLRP, USP. We thank Ricardo Alarcon and Mauricio de Oliveira for technical assistance.

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  1. Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes 3900, 14.040-901 Ribeirão Preto, São Paulo, Brazil

    A. C. M. M. Aquino, J. A. Jorge, H. F. Terenzi & M. L. T. M. Polizeli

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  1. A. C. M. M. Aquino
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  2. J. A. Jorge
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  3. H. F. Terenzi
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  4. M. L. T. M. Polizeli
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Correspondence to M. L. T. M. Polizeli.

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Aquino, A.C.M.M., Jorge, J.A., Terenzi, H.F. et al. Studies on a thermostable α-amylase from the thermophilic fungus Scytalidium thermophilum . Appl Microbiol Biotechnol 61, 323–328 (2003). https://doi.org/10.1007/s00253-003-1290-y

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  • DOI: https://doi.org/10.1007/s00253-003-1290-y

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