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Purification and characterization of a prolyl endopeptidase isolated from Aspergillus oryzae

  • Biotechnology Methods
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A new fungal strain that was isolated from our library was identified as an Aspergillus oryzae and noted to produce a novel proly endopeptidase. The enzyme was isolated, purified, and characterized. The molecular mass of the prolyl endopeptidase was estimated to be 60 kDa by using SDS-PAGE. Further biochemical characterization assays revealed that the enzyme attained optimal activity at pH 4.0 with acid pH stability from 3.0 to 5.0. Its optimum temperature was 30 °C and residual activity after 30 min incubation at 55 °C was higher than 80 %. The enzyme was activated and stabilized by Ca2+ but inhibited by EDTA (10 mM) and Cu2+. The K m and k cat values of the purified enzyme for different length substrates were also evaluated, and the results imply that the enzyme from A. oryzae possesses higher affinity for the larger substrates. Furthermore, this paper demonstrates for the first time that a prolyl endopeptidase purified from A. oryzae is able to hydrolyze intact casein.

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Acknowledgments

Financial support from the National High Technology Research and Development Program of China (863 Program) (No. 2012AA022207), the National Key Basic Research and Development Program of China (973 Program) (No. 2011CB710800) and the Program of Introducing Talents of Discipline to Universities (111 Project) (111-2-06) are greatly appreciated.

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  1. State Key Laboratory of Food Science and Technology, the Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Ave, Wuxi, 214122, Jiangsu, People’s Republic of China

    Chao Kang, Xiao-Wei Yu & Yan Xu

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  1. Chao Kang
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  2. Xiao-Wei Yu
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  3. Yan Xu
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Correspondence to Xiao-Wei Yu or Yan Xu.

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Kang, C., Yu, XW. & Xu, Y. Purification and characterization of a prolyl endopeptidase isolated from Aspergillus oryzae . J Ind Microbiol Biotechnol 41, 49–55 (2014). https://doi.org/10.1007/s10295-013-1378-z

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  • DOI: https://doi.org/10.1007/s10295-013-1378-z

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