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In vitro Antioxidant, Cytotoxic and Antidiabetic Activity of Protein Hydrolysates Prepared from Chinese Pond Turtle (Chinemys reevesii)

Md. Serajul Islam1,2orcid tiny, Wang Hongxin1,2*orcid tiny, Habtamu Admassu3orcid tiny, Amer Ali Mahdi1,2orcid tiny, Ma Chaoyang2 and Fu An Wei4orcid tiny

1State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi-214122, Jiangsu province, PR China 

2National Engineering Research Center for Functional Food, Jiangnan University, 1800 Lihu Avenue, Wuxi-214122, Jiangsu province, PR China 

3Biotechnology and Bioprocessing Center of Excellence/Department of Food Process Engineering, College of Biological and Chemical Engineering, Addis Ababa Science and Technology University, Addis Ababa, Ethiopia  

4Guangxi Zhongtaikang Technology Industry Co., Ltd., Nanning-530029, Guangxi, PR China

Article history:

Received: 1 December 2020

Accepted: 15 July 2021

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Key words:

Chinese pond turtle, molecular mass profiles, antioxidant activity, antidiabetic capacity, lipid peroxide inhibition, cytotoxic effect


Research backgroundCardiovascular diseases and diabetes are the biggest causes of death globally. Bioactive peptides derived from many food proteins using enzymatic proteolysis and food processing have a positive impact on the prevention of these diseases. The bioactivity of Chinese pond turtle muscle proteins and their enzymatic hydrolysates has not received much attention, thus this study aims to investigate their antioxidant, antidiabetic and cytotoxic activities.

Experimental approachChinese pond turtle muscles were hydrolysed using four proteolytic enzymes (Alcalase, Flavourzyme, trypsin and bromelain) and the degrees of hydrolysis were measured. High-performance liquid chromatography (HPLC) was conducted to explore the amino acid profiles and molecular mass distribution of the hydrolysates. The antioxidant activities were evaluated using various in vitro tests, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), hydroxyl radical scavenging activity, reducing capacity, chelating Fe2+ and lipid peroxide inhibition activity. Antidiabetic activity was evaluated using α-amylase inhibition and α-glucosidase inhibition assays. Besides, cytotoxic effect of hydrolysates on human colon cancer (HT-29) cells was assessed.

Results and conclusionsThe amino acid composition of the hydrolysates revealed higher mass fractions of glutamic, aspartic, lysine, hydroxyproline and hydrophobic amino acids. Significantly highest inhibition of lipid peroxidation was achieved when hydrolysate obtained with Alcalase was used. Protein hydrolysate produced with Flavourzyme had the highest radical scavenging activity measured by DPPH (68.32 %), ABTS (74.12 %) and FRAP (A700 nm=0.300) assays, α-glucosidase (61.80 %) inhibition and cytotoxic effect (82.26 %) on HT-29 cell line at 550 μg/mL. Hydrolysates obtained with trypsin and bromelain had significantly highest (p<0.05) hydroxyl radical scavenging (92.70 %) and Fe2+ metal chelating (63.29 %) activities, respectively. The highest α-amylase (76.89 %) inhibition was recorded when using hydrolysates obtained with bromelain and Flavourzyme.

Novelty and scientific contributionEnzymatic hydrolysates of Chinese pond turtle muscle protein had high antioxidant, cytotoxic and antidiabetic activities. The findings of this study indicated that the bioactive hydrolysates or peptides from Chinese pond turtle muscle protein can be potential ingredients in pharmaceuticals and functional food formulations.

*Corresponding author: +(086)51085917795

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