周勰 叶捷娜 张林佳 黄芳芳 杨最素

摘要 [目的]探讨胰蛋白酶制备乌贼缠卵腺抗氧化活性多肽的工艺及体外抗氧化活性。[方法]以乌贼缠卵腺为原料，用胰蛋白酶进行酶解，采用单因素和正交试验优化乌贼缠卵腺抗氧化活性肽的提取工艺;将酶解液超滤分成4个分子段，通过体外检测DPPH清除率、·OH清除率和还原力评价其抗氧化活性。[结果]胰蛋白酶酶解乌贼缠卵腺在加酶量2 000 U/g、pH为8.5、料液比1∶3、酶解时間8 h条件下得到的酶解物对DPPH的清除率最大。酶解液经超滤得到的<3 kD分子段的多肽具有良好的DPPH清除能力、·OH清除能力和还原能力。[结论]胰蛋白酶酶解制备的乌贼缠卵腺多肽具有较好的体外抗氧化活性，可作为抗氧化肽的理想来源。

关键词 乌贼缠卵腺;多肽;胰蛋白酶;酶解;抗氧化活性

中图分类号 TS 254.9  文献标识码 A

文章编号 0517-6611（2021）16-0183-04

doi：10.3969/j.issn.0517-6611.2021.16.049   开放科学（资源服务）标识码（OSID）：

Study on Preparation of Antioxidant Activity Polypeptide from Cuttlefish Ovary Gland by Trypsin

ZHOU Xie，YE Jie-na，ZHANG Lin-jia et al （Zhejiang Engineering Research Center for Marine Biomedical Products，School of Food and Pharmacy，Zhejiang Ocean University，Zhoushan，Zhejiang 316022）

Abstract [Objective] To explore the conditions for the preparation of antioxidant peptides from the nidamental glands of cuttlefish by trypsin and study the antioxidant activities of the enzymatic hydrolysates  in vitro. [Method] The nidamental glands of cuttlefish were used as raw materials and hydrolyzed by trypsin.Single-factor and orthogonal tests were used to optimize the extraction process for anti-oxidant peptides from the nidamental glands of cuttlefish.The enzyme hydrolysate was ultrafiltered into 4 molecular segments，and its antioxidant activity was evaluated by  in vitro  detection of DPPH clearance rate，·OH clearance rate and reducing power.[Result] The optimized conditions were as follows： volume of enzyme of 2 000 U/g，pH of 8.5，solid-liquid ratio of 1∶3 and hydrolysis time of 8 h.Peptides with a Mw of less than 3 kD exhibited appreciable antioxidant activities，including DPPH scavenging，·OH scavenging and reducing activities.[Conclusion]The peptides from the nidamental glands of cuttlefish by trypsin had excellent antioxidant activities  in vitro  and could be an ideal source of antioxidant peptides.

Key words Nidamental gland of cuttlefish;Peptides;Trypsin;Enzymatic hydrolysis;Antioxidant activity

人体内自由基的生成和清除一般处于动态平衡状态，过多的自由基会干扰这种平衡，从而产生氧化应激反应并引起炎症、糖尿病等疾病[1]。化学合成的抗氧化剂已普遍应用于生物医学产品、日用化妆品和食品等行业[2]。然而化学合成的抗氧化剂对人体健康有潜在的毒性，因此，开发安全有效的天然抗氧化剂已引起人们的广泛关注[3]。近年来，越来越多动植物来源的多肽已被证实具有清除自由基的能力[4]，这为开发新型抗氧化剂提供了新的思路。研究表明，通过酶解大分子蛋白得到的抗氧化肽通常具有抗氧化活性强、生物安全性等优势[5]。海洋蛋白是制备抗氧化活性肽的高质量原料。鲍鱼内脏、鲤鱼鱼鳞等海洋生物蛋白已被报道用于制备具有良好抗氧化活性的多肽[6-7]。……