Hu Xiao, Liu Jing, Li Jun, Song Yuqiong, Chen Shengjun, Zhou Shaobo, Yang Xianqing
Key Laboratory of Aquatic Product Processing, Ministry of Agriculture and Rural, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.
Co-innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Ocean University, Lianyungang, China.
Front Nutr. 2022 Jul 22;9:971419. doi: 10.3389/fnut.2022.971419. eCollection 2022.
() protein was hydrolyzed with alkaline protease to obtain antioxidant peptides. The enzymatic hydrolysis conditions were optimized through single-factor and orthogonal experiments. The results showed that the optimal process parameters were using 2% of alkaline protease, and substrate concentration of 1 g/100 mL and hydrolyzed 2 h at pH 8.0. Gel filtration chromatography and RP-HPLC were adopted for isolating and purifying the antioxidant peptides from the protein hydrolysate (GLPH). Three novel antioxidant peptides were identified as LSPGEL (614.68 Da), VYFDR (698.76 Da), and PGPTY (533.57 Da) by nano-HPLC-MS/MS. The results of ABTS free radical scavenging rate demonstrated PGPTY exhibited the best antioxidant activity (IC = 0.24 mg/mL). Moreover, LSPGEL, VYFDR, and PGPTY were docked with Keap1, respectively. The molecular docking results suggested PGPTY had smaller docking energy and inhibition constants than the other two peptides. Finally, the cell viability assay evidenced the protective effect exerted by the antioxidant peptide on HO-induced oxidative damage. Above findings showed the potential of using antioxidant peptides from GLPH as antioxidants.
（）蛋白用碱性蛋白酶水解以获得抗氧化肽。通过单因素和正交实验优化酶解条件。结果表明，最佳工艺参数为使用2%的碱性蛋白酶，底物浓度为1 g/100 mL，在pH 8.0下酶解2 h。采用凝胶过滤色谱和反相高效液相色谱从蛋白水解物（GLPH）中分离纯化抗氧化肽。通过纳米高效液相色谱-串联质谱鉴定出三种新型抗氧化肽，分别为LSPGEL（614.68 Da）、VYFDR（698.76 Da）和PGPTY（533.57 Da）。ABTS自由基清除率结果表明PGPTY表现出最佳的抗氧化活性（IC = 0.24 mg/mL）。此外，LSPGEL、VYFDR和PGPTY分别与Keap1进行对接。分子对接结果表明PGPTY的对接能量和抑制常数比其他两种肽小。最后，细胞活力测定证明了抗氧化肽对HO诱导的氧化损伤具有保护作用。上述研究结果表明GLPH中的抗氧化肽具有作为抗氧化剂的潜力。
