Antioxidant and cytotoxic properties of protein hydrolysates obtained from enzymatic hydrolysis of Klunzinger’s mullet (Liza klunzingeri) muscle

Sana Rabiei,Samira Asgharzade,Mahmoud Rafieia-Kopai,Masoud Rezaei,Mehdi Nikoo

doi:10.1590/s2175-97902019000218304

Abstract

Today, consumers are looking for functional foods that promote health and prevent certain diseases in addition to provide nutritional requirements. This study aimed to evaluate the antioxidant and cytotoxic properties of Liza klunzingeri protein hydrolysates. Fish protein hydrolysates (FPHs) were prepared from L. klunzingeri muscle using enzymatic hydrolysis with papain at enzyme/substrate ratios of 1:25 and 1:50 for 45, 90 and 180 min. The antioxidant activities of the FPHs were investigated through five antioxidant assays. The cytotoxic effects on 4T1 carcinoma cell line were also evaluated. The amino acid composition and molecular weight distribution of the hydrolysate with the highest antioxidant activity were determined by HPLC. All six FPHs exhibited good scavenging activity on ABTS (IC50=0.60-0.12 mg/mL), DPPH (IC50= 3.18-2.08 mg/mL), and hydroxyl (IC50=4.13-2.07 mg/mL) radicals. They also showed moderate Fe+2 chelating capacity (IC50=2.12-12.60 mg/mL) and relatively poor ferric reducing activity (absorbance at 70 nm= 0.01-0.15, 5 mg/mL). In addition, all hydrolysates showed cytotoxic activities against the 4T1 cells (IC50=1.62-2.61 mg/mL). 94.6% of peptide in hydrolysate with the highest antioxidant activity had molecular weight less than 1,000 Da. L. klunzingeri protein hydrolysates show significant antioxidant and anticancer activities in vitro and are suggested to be used in animal studies.

Highlights

In biological systems, free radicals are typically derived from the oxygen, nitrogen, and sulfur molecules
2,2’-azinobios-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferrozine (3-(2-Pyridyl)-5,6-diphenyl-1,2,4-triazine4′,4′′-disulfonic acid sodium salt), Dimethyl sulfoxide (DMSO), 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2Htetrazolium bromide (MTT), Dulbecco’s Modified Eagle’s Medium (DMEM), Fetal Bovine Serum (FBS), Butylated hydroxytoluene (BHT), and Trypsin-EDTA solution were purchased from Sigma-Aldrich Chemical Co
Protein hydrolysates obtained by enzymatic hydrolysis of L. kludingeri muscle using two concentration of papain at 45, 90, and 180 min, exhibited good scavenging activity on DPPH (IC50=2.08-3.18 mg/mL), ABTS (IC50=0.12-0.60 mg/mL), and hydroxyl (IC50 = 2.07‐4.13 mg/mL) radicals, moderate chelating activities on Fe2+ (IC50=2.12-12.60 mg/mL), and relatively poor ferric reducing activities

Summary

Introduction

Free radicals are typically derived from the oxygen, nitrogen, and sulfur molecules. Due to their unpaired electrons, free radicals exhibit a great deal of combination desire for reaction with other molecules. The most important free radical including Reactive Oxygen and Nitrogen Species (ROS/RNS) are naturally produced by various metabolic pathways such as the aerobic metabolism in mitochondrial respiratory chain, and play numerous physiological roles such as intracellular signaling, regulation of cell proliferation. The ROS and RNS radicals react with nucleic acids, the side chains of amino acid in proteins, and double bonds of unsaturated fatty acids, triggering and developing oxidative stress, which plays an important role in the pathogenesis of many human diseases, including cancer (Nikoo, Benjakul, 2015).

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