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摘要[目的] 为鱿鱼肝脏的高值化利用提供基础研究数据。[方法] 以鱿鱼肝脏蛋白为原料，通过蛋白酶水解来制备生物活性肽，研究了多种蛋白酶水解鱿鱼肝脏制得水解液的抗凝血活性、抑制血管紧张素I转换酶（ACE）活性及抗氧化活性。[结果] 当鱿鱼肝脏水解液浓度为5 mg/ml时，中性蛋白酶的水解液对ACE抑制率为68.0%；APTT值为59.8 s，TT值为34.2 s，比空白对照组分别延长26.7和20.6 s，表明鱿鱼肝脏的中性蛋白酶水解液具有抗凝血活性。中性蛋白酶解液对Fe3+的还原力为0.588，羟自由基清除能力为645%，说明鱿鱼肝脏中性蛋白酶水解液具有较好的抗氧化活性；而其余的木瓜蛋白酶、酸性蛋白酶、碱性蛋白酶、胰蛋白酶的鱿鱼肝脏水解液的抗凝血活性和ACE抑制活性均低于中性蛋白酶水解液的酶解液活性。[结论]鱿鱼肝脏蛋白质具有较好的综合利用价值。
关键词鱿鱼肝脏；生物活性肽；血管紧张素I转换酶；抑制活性；抗凝血活性
中图分类号文献标识码A文章编号0517-6611（2014）15-04666-03
Abstract[Objective] The research aimed to provide basic research data for highvalue utilization of squid liver. [Method] Taking protein of squid liver as materials， the bioactive peptide was prepared by protease hydrolysis. The anticoagulant activity， inhibitory activity and antioxidant activity of angiotensin converting enzyme I of the squid liver hydrolyzed with many kinds of protease were studied. [Result] When the hydrolyzate concentration of squid liver was 5 mg/ml， the inhibition rate of neutral protease hydrolyzate to ACE was 68.0%. APTT and TT value was 59.8 and 34.2 s respectively， which were 26.7 s and 20.6 s longer than the blank control respectively. This indicated that the neutral protease hydrolyzate of squid liver had anticoagulant activity. The reducing power of neutral protease hydrolyzate to Fe3+ was 0.588， the scavenging capacity of hydorxyl radical was 64.5%， which indicated neutral protease hydrolyzate of squid liver had higher antioxidant activity. The anticoagulant activity and ACE inhibitory activity of hydrolyzate of squid liver by papain， acid protease， alkaline protease and trypsase were lower than the activity of neutral protease hydrolyzate. [Conclusion] Squid liver protein had better comprehensive utilization value.
Key wordsSquid liver； Bioactive peptide； Angiotensin converting enzyme I； Inhibitory activity； Anticoagulant activity
近年来，我国鱿鱼渔获量占总渔获量的比重呈不断上升的趋势。在加工过程中产生大量副产物，这部分肝脏大都用于加工鱼粉、鱿鱼浆等初级产品或直接丢弃，这不仅造成资源浪费还对环境造成污染。鱿鱼肝脏约占鱿鱼湿重的15%左右，其中约含19%的蛋白质[1]。目前对于鱿鱼肝脏的研究主要集中在脂肪和蛋白质方面，其中蛋白质主要是制备酱油等调味品和酶制剂[2-3]，对于活性肽的研究报道较为少见。笔者以鱿鱼肝脏蛋白为原料，通过酶解技术制备生物活性肽，并对其生物活性进行研究，以期为生物活性肽的制备提供新途径。
1材料与方法
1.1试验材料
1.1.1原料与试剂。鱿鱼内脏购于辽宁大连海洋渔业总公司；木瓜蛋白酶、酸性蛋白酶、中性蛋白酶、碱性蛋白酶、胰蛋白酶，购于天津诺奥科技发展有限公司；胃蛋白酶，购于上海莲冠生物化学有限公司；α葡萄糖苷酶、对硝基苯麦芽戊糖（pNPG），购于Sigma公司；TT试剂、PT试剂、APTT试剂，购于上海太阳生物技术有限公司。
1.1.2仪器与设备。Multiskan Ascent酶标仪，购于巩义市予华仪器有限责任公司；凝血仪为德国TECO产品；高效液相色谱为大连依利特科学仪器有限公司产品。
1.2方法
1.2.1肝脏基本营养成分的测定。将-18 ℃冷冻的鱿鱼内脏，于室温解冻，分离鱿鱼肝脏洗净后对其基本营养成分（水分、脂肪、蛋白质含量、灰分）进行测定。
关键词鱿鱼肝脏；生物活性肽；血管紧张素I转换酶；抑制活性；抗凝血活性
中图分类号文献标识码A文章编号0517-6611（2014）15-04666-03
Abstract[Objective] The research aimed to provide basic research data for highvalue utilization of squid liver. [Method] Taking protein of squid liver as materials， the bioactive peptide was prepared by protease hydrolysis. The anticoagulant activity， inhibitory activity and antioxidant activity of angiotensin converting enzyme I of the squid liver hydrolyzed with many kinds of protease were studied. [Result] When the hydrolyzate concentration of squid liver was 5 mg/ml， the inhibition rate of neutral protease hydrolyzate to ACE was 68.0%. APTT and TT value was 59.8 and 34.2 s respectively， which were 26.7 s and 20.6 s longer than the blank control respectively. This indicated that the neutral protease hydrolyzate of squid liver had anticoagulant activity. The reducing power of neutral protease hydrolyzate to Fe3+ was 0.588， the scavenging capacity of hydorxyl radical was 64.5%， which indicated neutral protease hydrolyzate of squid liver had higher antioxidant activity. The anticoagulant activity and ACE inhibitory activity of hydrolyzate of squid liver by papain， acid protease， alkaline protease and trypsase were lower than the activity of neutral protease hydrolyzate. [Conclusion] Squid liver protein had better comprehensive utilization value.
Key wordsSquid liver； Bioactive peptide； Angiotensin converting enzyme I； Inhibitory activity； Anticoagulant activity
近年来，我国鱿鱼渔获量占总渔获量的比重呈不断上升的趋势。在加工过程中产生大量副产物，这部分肝脏大都用于加工鱼粉、鱿鱼浆等初级产品或直接丢弃，这不仅造成资源浪费还对环境造成污染。鱿鱼肝脏约占鱿鱼湿重的15%左右，其中约含19%的蛋白质[1]。目前对于鱿鱼肝脏的研究主要集中在脂肪和蛋白质方面，其中蛋白质主要是制备酱油等调味品和酶制剂[2-3]，对于活性肽的研究报道较为少见。笔者以鱿鱼肝脏蛋白为原料，通过酶解技术制备生物活性肽，并对其生物活性进行研究，以期为生物活性肽的制备提供新途径。
1材料与方法
1.1试验材料
1.1.1原料与试剂。鱿鱼内脏购于辽宁大连海洋渔业总公司；木瓜蛋白酶、酸性蛋白酶、中性蛋白酶、碱性蛋白酶、胰蛋白酶，购于天津诺奥科技发展有限公司；胃蛋白酶，购于上海莲冠生物化学有限公司；α葡萄糖苷酶、对硝基苯麦芽戊糖（pNPG），购于Sigma公司；TT试剂、PT试剂、APTT试剂，购于上海太阳生物技术有限公司。
1.1.2仪器与设备。Multiskan Ascent酶标仪，购于巩义市予华仪器有限责任公司；凝血仪为德国TECO产品；高效液相色谱为大连依利特科学仪器有限公司产品。
1.2方法
1.2.1肝脏基本营养成分的测定。将-18 ℃冷冻的鱿鱼内脏，于室温解冻，分离鱿鱼肝脏洗净后对其基本营养成分（水分、脂肪、蛋白质含量、灰分）进行测定。