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Purification and Characterization of a Chymosin from Rhizopus microsporus var. rhizopodiformis

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Abstract

Purification and characterization of a chymosin from Rhizopus microsporus var. rhizopodiformis were investigated in the present study. A newly isolated R. microsporus var. rhizopodiformis F518 produced a high level of milk-clotting activity (1,001 SU/mL). A chymosin from the fungus was purified 3.66-fold with a recovery yield of 33.2 %. The enzyme appeared as a single protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular mass of 37.0 kDa. It was optimally active at 60 °C and was stable up to 40 °C. The purified enzyme was an acid protease with an optimum pH of 5.2 and retained 80 % of residual activity within pH 2.0–8.0. The inhibition of 96 and 100 % by pepstatin A at 0.01 and 0.02 mM, respectively, revealed that the enzyme is an aspartic protease. Thus, high milk-clotting activity of the chymosin with good stability will strengthen the potential use of the chymosin as a substitute for calf rennet in cheese manufacturing.

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Acknowledgments

This work was supported by the National Science Fund for Distinguished Young Scholars (no. 31325021) and the Program for Changjiang Scholars and Innovative Research Team in University (no. IRT1293).

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Authors and Affiliations

  1. College of Food Science and Nutritional Engineering, Beijing Key Laboratory of Functional Food from Plant Resources, China Agricultural University, PO Box 294, No.17 Qinghua Donglu, Haidian District, Beijing, 100083, China

    Qian Sun & Zheng-Qiang Jiang

  2. College of Engineering, China Agricultural University, PO Box 294, No.17 Qinghua Donglu, Haidian District, Beijing, 100083, China

    Xi-Ping Wang & Qiao-Juan Yan

  3. College of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China

    Wei Chen

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  1. Qian Sun
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Correspondence to Qiao-Juan Yan or Zheng-Qiang Jiang.

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Sun, Q., Wang, XP., Yan, QJ. et al. Purification and Characterization of a Chymosin from Rhizopus microsporus var. rhizopodiformis . Appl Biochem Biotechnol 174, 174–185 (2014). https://doi.org/10.1007/s12010-014-1044-6

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