基于亲和色谱的海蜇生殖腺ACE抑制肽分离及其活性研究
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- 英文论文题名：Preparation of angiotensin converting enzyme(ACE) inhibitory peptides from jellyfish gonad hydrolysate based on affinity chromatography
- 论文作者：孙美玲 ; 张钦 ; 年益营 ; 马倩 ; 付颖寰
- 英文论文作者：Sun Meiling ; Zhang Qin ; Nian Yiying ; Ma Qian ; Fu Yinghuan ; School of Food Science and Technology ; Dalian Polytechnic University ; National Engineering Research Center of Seafood ; Dalian Polytechnic University ; School of Light Industry and Chemical Engineering ; Dalian Polytechnic University
- 年：2016
- 作者机构：大连工业大学食品学院;国家海洋食品工程技术研究中心;大连工业大学轻工与化学工程学院;
- 论文关键词：SBA-15 ; Zn-SBA-15 ; 血管紧张素转化酶 ; 亲和分离
- 英文论文关键词：SBA-15 ; Zn-SBA-15 ; angiotensin converting enzyme ; affinity separation
- 会议召开时间：2016-11-09
- 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
- 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
- 语种：中文
- 分类号：TS254.9;TS201.2
- 学会代码：ZGSP
- 会议名称：中国食品科学技术学会第十三届年会
- 会议地点：中国北京
- 主办单位：中国食品科学技术学会
- 学会名称：中国食品科学技术学会
- 页数：2
- 文件大小：106k
- 原文格式：O
- 会议级别：全国
摘要
以三嵌段共聚物P123为模板剂,通过水热法了制备介孔分子筛SBA-15并进行了XRD、N_2吸附表征。然后在其表面吸附Zn~(2+)制备亲和介质Zn~(2+)-SBA-15。采用硫酸铵分级沉淀法从猪肺中粗提得到血管紧张素转化酶ACE。运用亲和吸附原理对粗提血管紧张素转化酶进行固定化,并对影响固定化酶活的因素进行优化。得出最佳反应条件为:ACE蛋白质量浓度为55 g/L、温度为50℃、pH=8.8、反应60 rnin,在此条件下固定化ACE酶活为0.317 U/g。采用制得的固定化ACE对海蜇性腺酶解液中的ACE抑制肽(ACEIP)进行亲和分离,然后获得吸附肽的固定化酶载体,再通过提高盐浓度将ACEIP洗脱下来。测定海蜇性腺酶解液的蛋白浓度为25.1195g/L时的ACE抑制率是82.25%,所分离得到的ACEIP提取液蛋白浓度为0.5368g/L的ACE抑制率时31.58%。说明采用该固定化ACE对酶解液中的ACEIP进行分离纯化有明显的效果。
Mesoporous molecular sieve SBA-15 was prepared through hydrothermal method with three block copolymer P123 as the template agent and characterized by FTIR,N2-adsorption,XRD,TEM.Then adsorption of Zn~(2+) was done on the surface of the affinity medium to obtain Zn~(2+)-SBA-15.Angiotensin converting enzyme(ACE) was extracted by ammonium sulfate precipitation from pig's lung.Using Zn~(2+)-SBA-15 as a carrier,ACE was immobilized by adsorption and chelation.The factors influencing the activity of immobilized enzyme were optimized.The optimal reaction conditions were obtained as:ACE mass protein concentration of 55 g/L,the temperature 50 C,pH=8.8,immobilization time of 1 h,with an optimal specific activity of 0.317 U/g.ACE inhibitory peptides(ACEIP) were separated from jellyfish gonad hydrolysate by using immobilized ACE affinity chromatography,contain first adsorbed on the carrier and then eluted by improving the salt concentration.The ACE inhibitory activity of jellyfish gonad hydrolysate was 82.25%(which protein concentration is 25.1195 g/L),after separation,the ACE inhibitory activity was 31.58%(which protein concentration is 0.5368 g/L).These data preliminary display that using immobilized ACE for affinity separation and purification of ACEIP in marine protein hydrolysate has obvious effect and prospect.
Mesoporous molecular sieve SBA-15 was prepared through hydrothermal method with three block copolymer P123 as the template agent and characterized by FTIR,N2-adsorption,XRD,TEM.Then adsorption of Zn~(2+) was done on the surface of the affinity medium to obtain Zn~(2+)-SBA-15.Angiotensin converting enzyme(ACE) was extracted by ammonium sulfate precipitation from pig's lung.Using Zn~(2+)-SBA-15 as a carrier,ACE was immobilized by adsorption and chelation.The factors influencing the activity of immobilized enzyme were optimized.The optimal reaction conditions were obtained as:ACE mass protein concentration of 55 g/L,the temperature 50 C,pH=8.8,immobilization time of 1 h,with an optimal specific activity of 0.317 U/g.ACE inhibitory peptides(ACEIP) were separated from jellyfish gonad hydrolysate by using immobilized ACE affinity chromatography,contain first adsorbed on the carrier and then eluted by improving the salt concentration.The ACE inhibitory activity of jellyfish gonad hydrolysate was 82.25%(which protein concentration is 25.1195 g/L),after separation,the ACE inhibitory activity was 31.58%(which protein concentration is 0.5368 g/L).These data preliminary display that using immobilized ACE for affinity separation and purification of ACEIP in marine protein hydrolysate has obvious effect and prospect.
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