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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Acknowledgments
We thank Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) for financial support and Prof. Dr. Carlos Alberto Labate from ESALQ-USP for the MS analysis. We also thank Maurício de Oliveira for technical assistance.
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Giraldo, M.A., da Silva, T.M., Salvato, F. et al. Thermostable invertases from Paecylomyces variotii produced under submerged and solid-state fermentation using agroindustrial residues. World J Microbiol Biotechnol 28, 463–472 (2012). https://doi.org/10.1007/s11274-011-0837-9
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DOI: https://doi.org/10.1007/s11274-011-0837-9