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Production and characterization of a collagenolytic serine proteinase by Penicillium aurantiogriseum URM 4622: A factorial study

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Abstract

A 24 full factorial design was used to identify the main effects and interactions of the initial medium pH, soybean flour concentration, temperature and orbital agitation speed on extracellular collagenase production by Penicillium aurantiogriseum URM4622. The most significant variables for collagenase production were soybean flour concentration and initial medium pH that had positive main effects, and temperature that had a negative one. Protein concentration in soybean flour revealed to be a significant factor for the production of a collagenase serine proteinase. The most favorable production conditions were found to be 0.75% soybean flour, pH 8.0, 200 rpm, and 28°C, which led to a collagenase activity of 164 U. The enzyme showed an optimum activity at 37°C and pH 9.0, was stable over wide ranges of pH and temperature (6.0 ∼ 10.0 and 25 ∼ 45°C, respectively) and was strongly inhibited by 10 mM phenylmethylsulphonylfluoride. The firstorder rate constants for collagenase inactivation in the crude extract, calculated from semi-log plots of the residual activity versus time, were used in Arrhenius and Eyring plots to estimate the main thermodynamic parameters of thermoinactivation (E* d = 107.4 kJ/mol and ΔH* d = 104.7 kJ/mol). The enzyme is probably an extracellular neutral serine collagenase effective on azocoll, gelatin and collagen decomposition.

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Authors and Affiliations

  1. Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco-UFPE, PE, Pernambuco, Brazil

    Carolina A. Lima, José L. Lima Filho, Maria G. Carneiro da Cunha & Ana L. F. Porto

  2. Department of Biochemistry, Federal University of Pernambuco-UFPE, PE, Pernambuco, Brazil

    José L. Lima Filho & Maria G. Carneiro da Cunha

  3. Department of Fundamental Chemistry, Federal University of Pernambuco-UFPE (UFPE), Pernambuco, Brazil

    Benício B. Neto

  4. Department of Chemical and Process Engineering, University of Genoa, Genoa, Italy

    Attilio Converti

  5. Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco-UFRPE, PE, Pernambuco, Brazil

    Ana L. F. Porto

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  1. Carolina A. Lima
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Correspondence to Ana L. F. Porto.

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Lima, C.A., Filho, J.L.L., Neto, B.B. et al. Production and characterization of a collagenolytic serine proteinase by Penicillium aurantiogriseum URM 4622: A factorial study. Biotechnol Bioproc E 16, 549–560 (2011). https://doi.org/10.1007/s12257-010-0247-0

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  • DOI: https://doi.org/10.1007/s12257-010-0247-0

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