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Stucture of the complex between Mucor pusillus pepsin and the key domain of κ-casein for site-directed mutagenesis: a combined molecular modeling and docking approach

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Abstract

In the structural-based mutagenesis of Mucor pusillus pepsin (MPP), understanding how κ-casein interacts with MPP is a great challenge for us to explore. Chymosin-sensitive peptide is the key domain of κ-casein that regulates milk clotting through the specific proteolytic cleavage of its peptide bond (Phe105-Met106) by MPP to produce insoluble para-κ-casein. Here, we built the model of this large peptide using molecular modeling technique. Docking study showed that MPP can accommodate the designed model with a favorable binding energy and the docked complex has proven to locally resemble the inhibitor-chymosin complex. The catalytic mechanism for the peptide model binding with MPP was explored in terms of substrate-enzyme interaction and property of contacting surface. Some critical amino acid residues in the substrate binding cleft have been identified as an important guide for further site-directed mutagenesis. Glu13 and Leu11 in the S3 region of MPP, predicted as the special mutation sites, were confirmed to retain clotting activity and decrease the proteolytic activity. These novel mutants may provide a promising application for improving cheese flavor.

Novel mutants of mucor pusillus pepsin having a promising application for improving cheese flavor were found by using molecular modeling technology.

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Acknowledgments

This work was supported by National 863 Program 2006AA10Z306, National Public Benefit Research (Agriculture) Foundation (200903043), China Postdoctoral Science Foundation funded project (20100471246), Natural Science Foundation of China (31071574), Natural Science Foundation for the Youth (21004028) and The Earmarked Fund for Modern Agro-industrial Technology Research Systems in China (Nycytx-05-02).

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Authors and Affiliations

  1. Center of Agro-food Technology, Northeast Agricultural Research Center of China, No. 1363 Cai-Yu Street, Changchun, Jilin Province, 130033, People’s Republic of China

    Tiezhu Li, Jinghui Wang, Yuqiu Li, Li Zhang, Li Zheng, Zhuolin Li & Zhennai Yang

  2. College of Biological and Agricultural Engineering, Jilin University, Changchun, 130025, People’s Republic of China

    Li Zhang, Li Zheng & Zhennai Yang

  3. State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University, Changchun, 130023, People’s Republic of China

    Quan Luo

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  1. Tiezhu Li
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  2. Jinghui Wang
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  3. Yuqiu Li
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  4. Li Zhang
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  5. Li Zheng
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  6. Zhuolin Li
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  7. Zhennai Yang
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Correspondence to Zhennai Yang or Quan Luo.

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Li, T., Wang, J., Li, Y. et al. Stucture of the complex between Mucor pusillus pepsin and the key domain of κ-casein for site-directed mutagenesis: a combined molecular modeling and docking approach. J Mol Model 17, 1661–1668 (2011). https://doi.org/10.1007/s00894-010-0869-3

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  • DOI: https://doi.org/10.1007/s00894-010-0869-3

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