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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2014, Vol. 34 Issue (3): 103-108    DOI: 10.13523/j.cb.20140315
技术与方法     
具有ACE抑制活性的霞水母酶解肽制备条件优化
贾梦蛟1,2, 刘睿1,2, 陆杰霖1,2, 朱蕴菡1,2, 王令充1,2, 王欣之1,2, 吴皓1,2, 鲁明明3
1. 南京中医药大学药学院 南京 210023;
2. 江苏省海洋药用生物资源研究与开发重点实验室 南京 210023;
3. 江苏南星药物有限责任公司 南京 210023
Study on Preparation of ACE Inhibitory Enzymatic Peptides of Cyanea
JIA Meng-jiao1,2, LIU Rui1,2, LU Jie-lin1,2, ZHU Yun-han1,2, WANG Ling-chong1,2, WANG Xin-zhi1,2, WU Hao1,2, LU Ming-ming3
1. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China;
2. Jiangsu Key Laboratory of Research and Development in Marin Bio-resource Pharmaceutics, Nanjing 210023, China;
3. Jiangsu Nanxing Pharmaceutical Co., Ltd, Nanjing 210023, China
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摘要: 目的:采用胰蛋白酶制备霞水母ACE抑制肽,并以响应面法优化酶解肽制备工艺,通过超滤分离获得具有ACE抑制作用的酶解肽活性部位。方法:单因素实验以ACE抑制率为指标,考察温度、酶解时间、pH、加酶量,进一步通过响应面法优化酶解工艺,以不同截留分子量的纤维膜进行分离。结果:胰蛋白酶水解霞水母制备ACE抑制肽的最佳工艺条件为:酶解温度:49.6℃,加酶量为0.69%,pH为7.8,酶解时间2.0h,酶解产物的平均ACE抑制率为56.79%,与预测值57.20%的相对误差为0.41%,经过超滤分离获得分子量<1k Da的酶解产物,ACE抑制活性最高,其IC50为 66.58μg/ml。结论:确定霞水母酶解肽的制备工艺,其中分子量<1kDa的部分为主要效应物质基础。
关键词: 霞水母ACE抑制肽响应面超滤酶解工艺    
Abstract: Objective: Trypsin was used to prepare ACE inhibitory peptides from Cyanea. Response surface method was used to optimize the ACE inhibitory peptides. The ultratiltration membrane was subjected to obtain ACE inhibitory segments of Cyanea enzymatic hydrolysate. Methods: According to the ACE inhibitiory rate, temperature,reaction time,pH and enzyme dosage were chosen to optimize the hydrolysis conditions,then response surface design was furtherly used to optimize the enzymatic technology.Results: The best conditions were as follows: hydrolysis temperature 49.6℃, enzyme dosage 0.69%, hydrolysis pH 7.8, hydrolysis time 2 hours. The ACE inhibitory rate of hydrolysis peptides at best conditions was 56.79%, compared with the theoretical value of 57.20%, the relative error was 0.41%. The mass segment of Cyanea enzymatic hydrolysate obtained by ultratiltration that molecular weight less than 1kDa had the highest ACE inhibitory activity,which IC50 value is 66.58μg/ml.Conclusion: The preparation of ACE inhibitory enzymatic peptides of Cyanea was determined and its material basis was the mass segment that molecular weight less than 1kDa.
Key words: Cyanea    ACE inhibitory peptides    Response surface analysis    Ultrafiltration    enzymatic hydrolysis
收稿日期: 2013-11-11 出版日期: 2014-03-25
ZTFLH:  Q815  
基金资助: 国家海洋公益性行业科研专项(201305007);江苏高校优势学科建设工程(ysxk-2010);国家高新技术研究发展计划(863计划)课题实施方案(SS2013AA090303)资助项目
通讯作者: 吴皓     E-mail: whao5795@vip.sina.com
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引用本文:

贾梦蛟, 刘睿, 陆杰霖, 朱蕴菡, 王令充, 王欣之, 吴皓, 鲁明明. 具有ACE抑制活性的霞水母酶解肽制备条件优化[J]. 中国生物工程杂志, 2014, 34(3): 103-108.

JIA Meng-jiao, LIU Rui, LU Jie-lin, ZHU Yun-han, WANG Ling-chong, WANG Xin-zhi, WU Hao, LU Ming-ming. Study on Preparation of ACE Inhibitory Enzymatic Peptides of Cyanea. China Biotechnology, 2014, 34(3): 103-108.

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https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140315        https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I3/103

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