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A high salt tolerant neutral protease from Aspergillus Oryzae: Purification, characterization and kinetic properties

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Abstract

A neutral high salt tolerant protease from Aspergillus oryzae CICIM F0899 which could be used for soy sauce production and other relevant applications under high-salt conditions was purified to homogeneity through ammonium sulfate precipitation, ion-exchange chromatography and gel filtration chromatography with overall recovery of 2%. Its molecular weight was estimated to be 50 kDa by SDS-PAGE. The optimum pH and temperature for activity of the extracellular protease of A. oryzae CICIM F0899 were shown to be between 7.0–9.0, and 50°C, respectively. The protease behaved high salt tolerance in 18% NaCl and retained 72% of initial activity after 14 days, indicating the high stability. The enzyme activity was inhibited by metal ions such as Al3+ and Ag+, and slightly activated by Mn2+ and Cu2+. A kinetic model incorporating the Debye-Hückel limiting law was proposed for A. oryzae CICIM F0899 protease hydrolysis of casein at ionic strength NaCl from 0.10 to 3.18 M. It was found that, with the higher ionic strength, the Michaelis constant K m of the protease monotonically increased while the turnover number k cat decreased in accordance with first order kinetic model. The high-salt tolerant protease has been demonstrated to be promising for the soy sauce production process.

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Authors and Affiliations

  1. Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, 214122, China

    D. Wang, Z. Y. Zheng, J. Feng, X. B. Zhan, L. M. Zhang, J. R. Wu & C. C. Lin

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  1. D. Wang
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  2. Z. Y. Zheng
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  3. J. Feng
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  4. X. B. Zhan
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  5. L. M. Zhang
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  6. J. R. Wu
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  7. C. C. Lin
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Correspondence to X. B. Zhan.

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Wang, D., Zheng, Z.Y., Feng, J. et al. A high salt tolerant neutral protease from Aspergillus Oryzae: Purification, characterization and kinetic properties. Appl Biochem Microbiol 49, 378–385 (2013). https://doi.org/10.1134/S0003683813040170

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  • DOI: https://doi.org/10.1134/S0003683813040170

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