Food and Fermentation Industries >

2022 , Vol. 48 >Issue 2: 71 - 78

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027737

Optimization of extraction and purification of cathepsin L from Litopenaeus vannamei muscle

  • TANG Zhendong ,
  • SHAO Haiyan ,
  • ZHANG Di ,
  • LIU Shucheng ,
  • JI Hongwu ,
  • XU Jie ,
  • TAI Minrui ,
  • SU Weiming ,
  • LIANG Shanhao ,
  • HE Xuan
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  • 1(College of Food Science and Technology, Guangdong Ocean University, Guangdong Provincial Key Laboratory of Aquatic Products Processing and Safety, Guangdong Province Engineering Laboratory for Marine Biological Products, Guangdong Provincial Engineering Technology Research Center of seafood, Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution, Zhanjiang 524088, China)
    2(Collaborative Innovation Center of Seafood Deep Processing, Dalian Polytechnic University, Dalian 116034, China)

Received date: 2021-04-16

  Revised date: 2021-05-18

  Online published: 2022-02-28

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Abstract

The purpose of the study was to optimize the extraction condition of cathepsin L from shrimp muscle (Litopenaeus vannamei), to isolate and purify, cathepsin L in extraction solutions, and to verify the effect of cathepsin L on the degradation of myofibril protein. The shrimp muscle was used as raw materials. The extraction condition of cathepsin L from muscle was optimized by single-factor tests, Plackett-Burman test and double-factor test. The cathepsin L in extraction solutions was isolated and purified by Tris-HCl buffer extraction, ammonium sulfate precipitation, Q-Seharose F.F anion exchange chromatography, SephacrylS-100 gel filtration chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The effect of cathepsin L on the degradation of myofibril protein was verified by the confirmatory experiment. The optimal extraction conditions of cathepsin L from shrimp muscle were determined as follows: buffer system containing Tris-HCl buffer of 40 mmol/L, L-Cys 6 mmol/L and PMSF 0.4 mmol/L, the ratio of solid to liquid at 1∶8(g∶mL), pH 6.0. Under the optimized conditions, the total enzyme activity value of cathepsin L increased to 378.36 U, and the yield of cathepsin L was 150.1 U/g. The specific activity of purified cathepsin L increased from 0.34 to 101.15 U/mg, the purification multiple was 298.28 times, the yield was 16.50%, and the molecular weight was 46.4 kDa. The confirmatory experiment further confirmed that the purified cathepsin L had significant influences on the degradation of actin and myosin heavy chain. cathepsin L had significant effects on the degradation of myofibril protein. It provides a theoretical basis for further study on the related properties of cathepsin L and its degradation mechanism in shrimp muscle.

Key words： Litopenaeus vannamei; cathepsin L; separation and purification; process optimization; protein degradation

Cite this article

TANG Zhendong , SHAO Haiyan , ZHANG Di , LIU Shucheng , JI Hongwu , XU Jie , TAI Minrui , SU Weiming , LIANG Shanhao , HE Xuan . Optimization of extraction and purification of cathepsin L from Litopenaeus vannamei muscle[J]. Food and Fermentation Industries, 2022 , 48(2) : 71 -78 . DOI: 10.13995/j.cnki.11-1802/ts.027737

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