Food and Fermentation Industries >

2022 , Vol. 48 >Issue 14: 93 - 99

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.030020

Purification and structure prediction of milk-clotting protease from Tenebrio molitor larvae

  • YANG Xiang ,
  • QIAO Haijun ,
  • YANG Xiaoli ,
  • WEN Pengcheng ,
  • WANG Yue ,
  • ZHANG Weibing ,
  • ZHANG Zhongming
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  • 1(College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China)
    2(College of Science, Gansu Agricultural University, Lanzhou 730070, China)
    3(Gansu Institute of Business and Technology, Lanzhou 730000, China)

Received date: 2021-11-12

  Revised date: 2021-11-28

  Online published: 2022-08-19

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Abstract

Tenebrio molitor larvae, as an abundant resource, has profuse protein content, and good application potential, however, there are few reports on milk-clotting protease from T. molitor larvae. This study purified a milk-clotting protease from guts in T. molitor larvae by column chromatography. By liquid chromatography tandem mass spectrometry and Edman degradation, amino acid sequence of the protease was obtained. And the physicochemical properties and structure were predicted and analyzed based on bioinformatics. The results showed that a milk-clotting protease with a molecular mass of 29.68 kDa and a milk-clotting activity of 173.8 SU/mg was obtained. The protease had a C/P ration of 104.7, an isoelectric point of 4.22, a hydrophobic amino acid of 43.37% and an instability index of 31.49, indicating that its physicochemical properties were stable. Compared with bovine chymosin and camel chymosin, their N-terminal sequence and secondary structure had great differences. Since the normalized B-factor value of 64.8% amino acid residues were negative, this protease had a good stability. This study provides a theoretical basis for the research of insect-derived milk-clotting protease.

Key words： Tenebrio molitor larvae; milk-clotting protease; separation and purification; physicochemical properties; structure

Cite this article

YANG Xiang , QIAO Haijun , YANG Xiaoli , WEN Pengcheng , WANG Yue , ZHANG Weibing , ZHANG Zhongming . Purification and structure prediction of milk-clotting protease from Tenebrio molitor larvae[J]. Food and Fermentation Industries, 2022 , 48(14) : 93 -99 . DOI: 10.13995/j.cnki.11-1802/ts.030020

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