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《集美大学学报（自然科学版）》[ISSN:1007-7405/CN:35-1186/N]

卷:

第24卷

期数:

2019年第3期

页码:

167-173

栏目:

水产、食品与生物工程

出版日期:

2019-05-28

文章信息/Info

Title:

Effect of Metal Ions on the Activity of a Prolyl Endopeptidase from Haliotis discus hannai Using Circular Dichroism Spectrum

作者:

李越1; 颜龙杰1; 翁凌1; 2; 章骞1; 2; 曹敏杰1; 2

（1.集美大学食品与生物工程学院，福建 厦门 361021；2.水产品深加工技术国家地方联合工程研究中心，福建 厦门 361021）

Author(s):

LI Yue1; YAN Longjie1; WENG Ling1; 2; ZHANG Qian1; 2; CAO Minjie1; 2

(1．College of Food and Biological Engineering，Jimei University，Xiamen 361021，China;2．National & Local Joint Engineering Research Center of Processing Technology for Aquatic Products，Xiamen 361021，China)

关键词:

脯氨酰内肽酶; 皱纹盘鲍; 金属离子; 圆二色谱; 抑制剂

Keywords:

prolyl-endopeptidase; Haliotis discus hannai; metal ions; circular dichroism spectrum; inhibitors

分类号:

-

DOI:

-

文献标志码:

A

摘要:

使用原核表达系统表达了皱纹盘鲍（Haliotis discus hannai）脯氨酰内肽酶（prolyl endopeptidase,PEP)，并对其进行高度纯化。SDS-PAGE结果表明，该酶的相对分子质量约为85 ku，其二级结构主要由α-螺旋（28.3%）、反向平行结构（17.4%）、平行结构（8.70%）、β转角（19.0%）、无规卷曲（28.5%）组成。选取PEP特异性荧光底物Suc-Gly-Pro-MCA测定其活性，并选取了多种常见金属盐溶液对其进行抑制作用分析。结果发现，Al3+、Cu2+、 Zn2+、Ag+ 和 Pb2+能够抑制PEP的活性。利用圆二色谱法，对金属离子作用下PEP的二级结构变化进行分析和计算。分析结果表明，不同金属离子对PEP的作用效果不同：Al3+、Cu2+和Zn2+ 在影响PEP结构时也影响其活性；Ca2+、Mg2+对PEP的结构和活性均不产生明显影响，而Ag+ 和 Pb2+可在不影响PEP二级结构的前提下抑制其活性。

Abstract:

Purified recombinant PEP from abalone (Haliotis discus hannai) was obtained using a prokaryotic expression system,with a relative molecular mass of 85 ku.The secondary structure of PEP mainly contained of alpha-helix-28.3%,antiparallel-17.4%,parallel-8.70%,beta-turn-19.0% and random coil-28.5%.The measurement of enzyme activity was based on a specific fluorogenic substrate Suc-Gly-Pro-MCA and a variety of common metal salts solution was selected for the inhibition assay.The results showed that the enzyme activity of PEP was strongly inhibited by Al3+,Cu2+,Zn2+,Ag+ and Pb2+.The secondary structure of PEP treated with metal ions was analyzed using circular dichroism spectrum.The influence of various metal ions on the PEP activity was different.In comparison,Al3+,Cu2+ and Zn2+ had an impact on both enzyme activity and the secondary structure,while Ca2+ and Mg2+ did not influence the secondary structure and activity obviously.Ag+ as well as Pb2+ revealed a strong inhibition on enzyme activity while less influence on the secondary structure.

参考文献/References:

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相似文献/References:

[1]赵春暖,严正凛.皱纹盘鲍与九孔鲍杂交的研究[J].集美大学学报(自然科学版),2014,19(5):339.
　ZHAO Chun-nuan,YAN Zheng-lin.Hybridization Between Haliotis discus hannai and Haliotis diversicolor aquatilis[J].Journal of Jimei University,2014,19(3):339.
[2]段雪昆,杜翠红,胡健健,等.皱纹盘鲍Hdh-MMP-1基因cDNA的克隆及原核表达[J].集美大学学报(自然科学版),2016,21(5):321.
　DUAN Xue-kun,DU Cui-hong,HU Jian-jian,et al.cDNA Cloning and Prokaryotic Expression of Matrix Metalloproteinase-1 from Haliotis discus hannai[J].Journal of Jimei University,2016,21(3):321.
[3]胡健健,陈玉磊,段雪昆,等.皱纹盘鲍亮氨酸氨肽酶基因cDNA克隆及表达分析[J].集美大学学报(自然科学版),2017,22(5):31.
　HU Jianjian,CHEN Yulei,DUAN Xuekun,et al.Fulllength cDNA Cloning and Expression Analysis of a Leucine Aminopeptidase from Haliotis discus hannai[J].Journal of Jimei University,2017,22(3):31.
[4]欧俊新,严正凛.南方鲍藻混养技术研究及其效益分析[J].集美大学学报(自然科学版),2011,16(3):172.
　OU Jun-xin,YAN Zheng-lin.The Technology and Efficiency of MacrophyteHaliotis Mixed Breeding Mode in South China[J].Journal of Jimei University,2011,16(3):172.
[5]万楚君,谢雪琼,张凌晶,等.温度和时间对鲍鱼消化的影响及产物的ACE抑制活性[J].集美大学学报(自然科学版),2019,24(4):258.
　WAN Chujun,XIE Xueqiong,ZHANG Lingjing,et al.Effects of Freezing Temperature and Duration on Digestion Characteristics of Abalone (Haliotis discus hannai) and Its ACE Inhibitory Activity[J].Journal of Jimei University,2019,24(3):258.
[6]陈玉磊,李婉玉,吴裕玲,等.皱纹盘鲍基质金属蛋白酶基因的克隆与表达分析[J].集美大学学报(自然科学版),2020,25(1):16.
　CHEN Yulei,LI Wanyu,WU Yuling,et al.Molecular Cloning and Expression Analysis of Matrix Metalloproteinases from Abalone(Haliotis discus hannai)[J].Journal of Jimei University,2020,25(3):16.

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更新日期/Last Update: 2019-06-17