Extracellular β-D-fructofuranosidase from Aspergillus parasiticus: Optimization of the production under submerged fermentation and biochemical characterization

  • André Luis Lucca
  • João Atílio Jorge
  • Luis Henrique Souza Guimarães

Abstract

The β-D-fructofuranosidases are enzymes with biotechnological potential that can be used in different industrial sectors as food and beverage. In this context, microorganisms are important producers of these biomolecules, especially filamentous fungi. The production of extracellular β-Dfructofuranosidase from Aspergillus parasiticus using sugarcane bagasse as a carbon source under submerged fermentation was optimized by factorial design and high levels of enzyme were obtained in 24 h-old cultures at 30°C using 1.5% sugarcane bagasse under agitation. The extracellular β-Dfructofuranosidase was purified 119-fold using diethyl aminoethyl (DEAE)-cellulose and Sephacryl S-200 chromatographic columns with recovery of 16%. The native molecular mass was estimated as 136 kDa with two subunits of 63 kDa determined by 7% sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) and 64% carbohydrate content. The purified enzyme showed optimal temperature of activity from 38-56°C and optimum pH from 4.5 to 6.2 determined by experimental design (CCRD), with half-life of 25 min at 50°C. It was stable from pH 5.0-10.0. The extracellular enzyme activity was stimulated by Ba2+ and Mg2+, and it was not affected by urea, silver and ethylenediaminetetraacetic acid (EDTA). The K0.5 and Vmax values were 10 mM and 1565 U/mg of protein, and 19 mM and 1965 U/mg of protein for sucrose and raffinose, respectively.

Keywords: Invertase, fructofuranosidase, Aspergillus parasiticus, sugarcane bagasse, factorial design.

African Journal of Biotechnology Vol. 12(38), pp. 5678-5687

Author Biographies

André Luis Lucca
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - USP Avenida Bandeirantes 3900, Monte Alegre, 14040-901 Ribeirão Preto, São Paulo, Brazil
João Atílio Jorge
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - USP Avenida Bandeirantes 3900, Monte Alegre, 14040-901 Ribeirão Preto, São Paulo, Brazil
Luis Henrique Souza Guimarães
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - USP Avenida Bandeirantes 3900, Monte Alegre, 14040-901 Ribeirão Preto, São Paulo, Brazil
Published
2016-05-16
Section
Articles

Journal Identifiers


eISSN: 1684-5315