Thermostable invertases from Paecylomyces variotii produced under submerged and solid-state fermentation using agroindustrial residues |
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Authors: | Marielle Aleixo Giraldo Tony Márcio da Silva Fernanda Salvato Héctor Francisco Terenzi Jo?o Atílio Jorge Luis Henrique Souza Guimar?es |
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Institution: | (1) Instituto de Qu?mica de Araraquara, UNESP, Rua Francisco Degni s/n, 14800-900 Araraquara, S?o Paulo, Brazil;(2) Departamento de Biologia, Faculdade de Filosofia, Ci?ncias e Letras de Ribeir?o Preto, USP, Avenida Bandeirantes 3900, 14040-901 Ribeir?o Preto, S?o Paulo, Brazil;(3) Departamento de Gen?tica, Escola Superior de Agricultura Luiz de Queiroz (ESALQ), USP, Avenida P?dua Dias 11, 13418-900 Piracicaba, S?o Paulo, Brazil; |
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Abstract: | The filamentous fungus Paecylomices variotii was able to produce high levels of cell extract and extracellular invertases when grown under submerged fermentation (SbmF)
and solid-state fermentation, using agroindustrial products or residues as substrates, mainly soy bran and wheat bran, at
40°C for 72 h and 96 h, respectively. Addition of glucose or fructose (≥1%; w/v) in SbmF inhibited enzyme production, while
the addition of 1% (w/v) peptone as organic nitrogen source enhanced the production by 3.7-fold. However, 1% (w/v) (NH4)2HPO4 inhibited enzyme production around 80%. The extracellular form was purified until electrophoretic homogeneity (10.5-fold
with 33% recovery) by DEAE-Fractogel and Sephacryl S-200 chromatography. The enzyme is a monomer with molecular mass of 102 kDa
estimated by SDS–PAGE with carbohydrate content of 53.6%. Optima of temperature and pH for both, extracellular and cell extract
invertases, were 60°C and 4.0–4.5, respectively. Both invertases were stable for 1 h at 60°C with half-lives of 10 min at
70°C. Mg2+, Ba2+ and Mn2+ activated both extracellular and cell extract invertases from P. variotii. The kinetic parameters Km and Vmax for the purified extracellular enzyme corresponded to 2.5 mM and 481 U/mg prot−1, respectively. |
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