NATURAL PRODUCT RESEARCH AND DEVELOPMENT ›› 2023, Vol. 35 ›› Issue (5): 819-827.doi: 10.16333/j.1001-6880.2023.5.010

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Optimization of extraction process of peroxidase from wolfberry and its enzymatic characteristics

GAO Qian-ling,HU Ling-wen, CHEN Hong-ping*,LIU You-ping,CHEN Lin,HU Yuan   

  1. State Key Laboratory of Southwest Characteristic Traditional Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China
  • Online:2023-05-26 Published:2023-05-26

Abstract:

Wolfberry is a commonly used Chinese medicinal material with “medicine and food homology”,which is prone to browning during processing,storage,and transportation,resulting in seriously affects the appearance and quality of Chinese wolfberry.Our previous study found that wolfberry has high peroxidase activities,and peroxidase (POD) plays an essential part in the enzymatic browning of plants.In order to better control the occurrence of enzymatic browning of wolfberry,the extraction process of peroxidase from wolfberry was optimized by response surface method,and the properties of POD from wolfberry were studied.The results showed that the optimal extraction process of POD was as follows:the solid-liquid ratio was 1∶3,the extraction time was 5 h,and the buffer pH was 6.0,the enzyme activity of POD was (5 148.59±50.00) U,which was similar to the predicted value.The extraction conditions of POD could be predicted and analyzed by the response surface model.The optimum temperature and pH were 50 ℃ and 6.0.The effects of different metal ions on the PPO were studied,and the result showed that Na+,Ca2+,K+,Zn2+,Cu2+ and citric acid could activate the activity of POD; ascorbic acid and sulfite could inhibit the activity of POD.The kinetics of enzymatic reaction was fit to Michaelis-Menten equation,and the Km value of the enzyme to guaiacol was 21.52 mmol/L and the Vmax was 115.47 U/mL.The Km value of H2O2 was 1.41 mmol/L and the Vmax was 148.81 U/mL.This study can provide a reference for the control of browning in the subsequent processing of wolfberry.

Key words: wolfberry, peroxidase, extraction, response surface methodology, enzymatic characteristics

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