酶解制备鱼鳞蛋白降血压肽的工艺优化
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摘要
为有效利用鱼鳞制备降血压肽,以罗非鱼鱼鳞为原料,在121℃条件下进行热预处理15 min后,运用响应面分析法优化酶解制备鱼鳞蛋白ACE抑制肽的工艺条件。结果表明,以水解度和ACE抑制率为评价指标,筛选出碱性蛋白酶为最优酶。在单因素试验的基础上,根据Box-Behnken中心组合试验设计原理,最终确立最优的酶解工艺参数为:酶解时间2 h、酶解温度56.3℃、pH值8.0,酶底比1.1%,此条件下ACE抑制率理论值为87.95%,实际值为88.26%。最优条件下制得的酶解产物相对分子质量集中在300～3 000 Da之间,水解效果较好。本研究结果对酶解法制备鱼鳞蛋白降血压活性肽具有一定的实践参考价值。
Thermally hydrolyzed fish scale protein hydrolysates demonstrated noticeable angiotensin converting enzyme(ACE) inhibitory activity. In order to utilize the tilapia scale to obtain ACE inhibitory peptide more effectively, the enzymatic hydrolysis conditions of tilapia scale protein were optimized using response surface methodology(RSM) after treated at 121℃ for 15 min. The results showed that alcalase was the more suitable enzyme for protein hydrolysis by comparing the degree of hydrolysis(DH) and ACE inhibition rate. Box-Behnken center-united experimental design was carried out based on single-factor experiments. The optimal parameters obtained were as follows, 2 h of hydrolysis time, 56.3℃ of hydrolysis temperature, 1.1% of enzyme-substrate ratio, and pH value 8.0. Under the optimal conditions, the theoretical value of ACE inhibition rate was 87.95%, while the actual value was 88.26%. The relative molecular mass of the hydrolyzate mainly ranged from 300 to 3000 Da. The results of this study provide a reference for the utilization use of fish scale protein to obtain hypotensive active peptide through enzymatic hydrolysis.
Thermally hydrolyzed fish scale protein hydrolysates demonstrated noticeable angiotensin converting enzyme(ACE) inhibitory activity. In order to utilize the tilapia scale to obtain ACE inhibitory peptide more effectively, the enzymatic hydrolysis conditions of tilapia scale protein were optimized using response surface methodology(RSM) after treated at 121℃ for 15 min. The results showed that alcalase was the more suitable enzyme for protein hydrolysis by comparing the degree of hydrolysis(DH) and ACE inhibition rate. Box-Behnken center-united experimental design was carried out based on single-factor experiments. The optimal parameters obtained were as follows, 2 h of hydrolysis time, 56.3℃ of hydrolysis temperature, 1.1% of enzyme-substrate ratio, and pH value 8.0. Under the optimal conditions, the theoretical value of ACE inhibition rate was 87.95%, while the actual value was 88.26%. The relative molecular mass of the hydrolyzate mainly ranged from 300 to 3000 Da. The results of this study provide a reference for the utilization use of fish scale protein to obtain hypotensive active peptide through enzymatic hydrolysis.
引文
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[12] 陈伟杰, 耿丽晶, 王馨云, 陈博, 周围. 菠萝皮蛋白酶酶解草鱼鱼鳞提取胶原蛋白的工艺优化[J]. 饲料研究, 2016(13):1-7
[13] 吴靖娜, 许永安, 王茵, 苏捷. 罗非鱼鱼鳞胶制备降血压胶原肽的工艺及酶解物活性[J]. 广东海洋大学学报, 2012, 32(1):75-80
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[17] 黄丹丹, 马良, 蔡路昀, 刘轶, 杨晖, 韩霜, 张宇昊. 预处理对酶解过程中胶原ACE抑制肽释放的影响[J]. 现代食品科技, 2016, 32(7):85-90
[18] Bougatef A, Nedjar-Arroume N, Ravallec-Plé R, Leroy Y, Guillochon D, Barkia A, Nasri M. Angiotensin Ⅰ-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases[J]. Food Chemistry, 2008, 111(2):350-356
[19] Wu Q Y, Du J J, Jia J Q, Kuang C. Production of ACE inhibitory peptides from sweet sorghum grain protein using alcalase: hydrolysis kinetic, purification and molecular docking study[J]. Food Chemistry, 2016, 199:140-149
[20] Qu W J, Ma H L, Pan Z L, Luo L,Wang Z B, He R H. Preparation and antihypertensive activity of peptides from Porphyra yezoensis[J]. Food Chemistry, 2010, 123(1):14-20
[21] 于丽娜, 齐宏涛, 张初署, 毕洁, 王明清, 杨伟强, 孙杰, 徐同城. 响应面法优化超声波辅助酶解制备花生蛋白抗菌肽[J]. 核农学报, 2018, 32(4):740-750
[22] 张周莉,李诚,刘爱平,徐晔,杨迪,朱娜,蔡婷婷. 响应面优化酶解法制备猪肩胛骨抗氧化肽工艺[J]. 核农学报, 2017, 31(12): 2358-2366
[23] 崔楠,陶晓赟,李娟,陈健,赵立仪,孙爱东. 响应面法优化黄粉虫蛋白制备ACE抑制肽的条件[J].食品科学, 2014, 35(15):156-160
[24] 温建丰,杨文鸽,徐大伦. 响应面法优化花蟹肉制备抗氧化肽的酶解工艺[J].核农学报, 2013, 27(12):1881-1886
[25] 胡杨,杨莉莉,熊善柏,刘友明,尤娟,尹涛. 不同预处理方法对酶法制备的草鱼鳞胶原蛋白肽特性的影响[J]. 华中农业大学学报, 2016, 35(5):105-111
[26] 王喜波, 王健, 张泽宇, 陈爽, 高婷婷, 江连洲. 物理改性对大豆蛋白柔性与乳化性的影响及其相关性分析[J]. 农业机械学报, 2017, 48(7):339-344
[27] Yang J Ⅰ, Ho H Y, Chu Y J, Chow C J. Characteristic and antioxidant activity of retorted gelatin hydrolysate from cobia (Rachycentron canadum) skin[J]. Food Chemistry, 2008, 110: 128-136
[28] Yang J Ⅰ, Liang W S, Chow C J, Siebert K J. Process for the production of tilapia retorted skin gelatin hydrolysates with optimized antioxidative properties[J]. Process Biochemistry, 2009, 44(10):1152-1157
[29] Min S G, Jo Y J, Hee Park S. Potential application of static hydrothermal processing to produce the protein hydrolysates from porcine skin by-products[J]. LWT-Food Science and Technolog, 2017, 83:18-25
[30] 王彦蓉, 崔春, 赵谋明, 欧阳珊, 肖如武. 罗非鱼鱼皮鱼鳞蛋白的酶解及超滤分离[J]. 食品与发酵工业, 2011, 37(9): 133-136
[31] 沙小梅, 郝君晖, 涂宗财, 胡姿姿, 王振兴, 张露, 李鑫, 王辉, 黄涛. 基于亚临界水技术的鱼骨软化及其在鱼糜中的应用[J].食品与发酵工业, 2018, 44(2):154-159
[32] 贺江, 赵自龙, 彭磊, 杨浩, 黄冰. 草鱼鱼鳞胶原蛋白提取及活性肽制备研究[J]. 食品研究与开发, 2017, 38(5): 52-55
[33] 张丰香, 李清华, 王霞, 周健. 鱼鳞血管紧张素转化酶(ACE)抑制肽的制备工艺研究[J]. 食品工业科技, 2013, 34(24): 199-204
[2] 操德群, 何艳丽, 余虹, 徐晓婷, 徐年军. 海洋生物ACE抑制肽研究进展[J]. 核农学报, 2017, 31(5): 927-937
[3] Honest N, Kessy E, Wang K, Zhao L, Zhou M L, Hu Z Y. Enrichment and biotransformation of phenolic compounds from litchi pericarps with angiotensin Ⅰ-converting enzyme (ACE) inhibition activity[J].LWT-Food Science and Technology,2018,87:301-309
[4] 王晓丹,薛璐,胡志和,张琦,李艳军,夏磊. ACE抑制肽构效关系研究进展[J].食品科学,2017, 38(5):305-310
[5] Lafarga T, Hayes M. Bioactive protein hydrolysates in the functional food ingredient industry: overcoming current challenges[J].Food Reviews International,2017, 33(3):217-246
[6] 林凯,韩雪,张兰威,谯飞,程大友. ACE抑制肽构效关系及其酶法制备的研究进展[J].食品科学, 2017, 38(3):261-270
[7] 曾健辉,朱宝君,王娟,孔繁茂,马广智,黄儒强. 响应面法优化酶解鱼鳞制备ACE抑制肽的研究[J].华南师范大学学报(自然科学版), 2016, 48(4):57-61
[8] 王南平,何兰,曹俊,郭休玉. 鱼鳞综合开发利用工艺与设备综述[J].渔业现代化, 2014, 41(1):46-52
[9] Gu R Z, Li C Y, Liu W Y, Yi W X, Cai M Y. Angiotensin Ⅰ-converting enzyme inhibitory activity of low-molecular-weight peptides from Atlantic salmon (Salmo salar L.) skin[J]. Food Research International, 2011, 44(5):1536-1540
[10] Thuanthong M, Gobba C D, Sirinupong N, Youravong W, Otte J. Purification and characterization of angiotensin-converting enzyme-inhibitory peptides from Nile tilapia (Oreochromis niloticus) skin gelatine produced by an enzymatic membrane reactor[J]. Journal of Functional Foods, 2017, 36:243-254
[11] 曾健辉,朱宝君,王娟,孔繁茂,马广智, 黄儒强. 响应面法优化酶解鱼鳞制备ACE抑制肽的研究[J].华南师范大学学报(自然科学版), 2016, 48(4):57-61
[12] 陈伟杰, 耿丽晶, 王馨云, 陈博, 周围. 菠萝皮蛋白酶酶解草鱼鱼鳞提取胶原蛋白的工艺优化[J]. 饲料研究, 2016(13):1-7
[13] 吴靖娜, 许永安, 王茵, 苏捷. 罗非鱼鱼鳞胶制备降血压胶原肽的工艺及酶解物活性[J]. 广东海洋大学学报, 2012, 32(1):75-80
[14] 中华人民共和国卫生部.GB/T 5009.5-2016食品中蛋白质的测定[S].北京:中国标准出版社,2016
[15] Spellman D, McEvoy E, O′Cuinn B G, FitzGerald R J. Proteinase and exopeptidase hydrolysis of whey protein: Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis[J]. International Dairy Journal, 2003, 13(3):447-453
[16] Wu J P, Aluko R E, Muir A D. Improved method for direct high performance liquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions[J]. Journal of Chromatography A, 2002, 950(1): 125-130
[17] 黄丹丹, 马良, 蔡路昀, 刘轶, 杨晖, 韩霜, 张宇昊. 预处理对酶解过程中胶原ACE抑制肽释放的影响[J]. 现代食品科技, 2016, 32(7):85-90
[18] Bougatef A, Nedjar-Arroume N, Ravallec-Plé R, Leroy Y, Guillochon D, Barkia A, Nasri M. Angiotensin Ⅰ-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases[J]. Food Chemistry, 2008, 111(2):350-356
[19] Wu Q Y, Du J J, Jia J Q, Kuang C. Production of ACE inhibitory peptides from sweet sorghum grain protein using alcalase: hydrolysis kinetic, purification and molecular docking study[J]. Food Chemistry, 2016, 199:140-149
[20] Qu W J, Ma H L, Pan Z L, Luo L,Wang Z B, He R H. Preparation and antihypertensive activity of peptides from Porphyra yezoensis[J]. Food Chemistry, 2010, 123(1):14-20
[21] 于丽娜, 齐宏涛, 张初署, 毕洁, 王明清, 杨伟强, 孙杰, 徐同城. 响应面法优化超声波辅助酶解制备花生蛋白抗菌肽[J]. 核农学报, 2018, 32(4):740-750
[22] 张周莉,李诚,刘爱平,徐晔,杨迪,朱娜,蔡婷婷. 响应面优化酶解法制备猪肩胛骨抗氧化肽工艺[J]. 核农学报, 2017, 31(12): 2358-2366
[23] 崔楠,陶晓赟,李娟,陈健,赵立仪,孙爱东. 响应面法优化黄粉虫蛋白制备ACE抑制肽的条件[J].食品科学, 2014, 35(15):156-160
[24] 温建丰,杨文鸽,徐大伦. 响应面法优化花蟹肉制备抗氧化肽的酶解工艺[J].核农学报, 2013, 27(12):1881-1886
[25] 胡杨,杨莉莉,熊善柏,刘友明,尤娟,尹涛. 不同预处理方法对酶法制备的草鱼鳞胶原蛋白肽特性的影响[J]. 华中农业大学学报, 2016, 35(5):105-111
[26] 王喜波, 王健, 张泽宇, 陈爽, 高婷婷, 江连洲. 物理改性对大豆蛋白柔性与乳化性的影响及其相关性分析[J]. 农业机械学报, 2017, 48(7):339-344
[27] Yang J Ⅰ, Ho H Y, Chu Y J, Chow C J. Characteristic and antioxidant activity of retorted gelatin hydrolysate from cobia (Rachycentron canadum) skin[J]. Food Chemistry, 2008, 110: 128-136
[28] Yang J Ⅰ, Liang W S, Chow C J, Siebert K J. Process for the production of tilapia retorted skin gelatin hydrolysates with optimized antioxidative properties[J]. Process Biochemistry, 2009, 44(10):1152-1157
[29] Min S G, Jo Y J, Hee Park S. Potential application of static hydrothermal processing to produce the protein hydrolysates from porcine skin by-products[J]. LWT-Food Science and Technolog, 2017, 83:18-25
[30] 王彦蓉, 崔春, 赵谋明, 欧阳珊, 肖如武. 罗非鱼鱼皮鱼鳞蛋白的酶解及超滤分离[J]. 食品与发酵工业, 2011, 37(9): 133-136
[31] 沙小梅, 郝君晖, 涂宗财, 胡姿姿, 王振兴, 张露, 李鑫, 王辉, 黄涛. 基于亚临界水技术的鱼骨软化及其在鱼糜中的应用[J].食品与发酵工业, 2018, 44(2):154-159
[32] 贺江, 赵自龙, 彭磊, 杨浩, 黄冰. 草鱼鱼鳞胶原蛋白提取及活性肽制备研究[J]. 食品研究与开发, 2017, 38(5): 52-55
[33] 张丰香, 李清华, 王霞, 周健. 鱼鳞血管紧张素转化酶(ACE)抑制肽的制备工艺研究[J]. 食品工业科技, 2013, 34(24): 199-204