[1]李越,颜龙杰,翁凌,等.圆二色谱法分析金属离子对鲍鱼脯氨酰内肽酶的作用[J].集美大学学报(自然版),2019,24(3):167-173.
 LI Yue,YAN Longjie,WENG Ling,et al.Effect of Metal Ions on the Activity of a Prolyl Endopeptidase from Haliotis discus hannai Using Circular Dichroism Spectrum[J].Journal of Jimei University,2019,24(3):167-173.
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圆二色谱法分析金属离子对鲍鱼脯氨酰内肽酶的作用()
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《集美大学学报(自然版)》[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、食品与生物技术
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12章骞12曹敏杰12
(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 Yue1YAN Longjie1WENG Ling12ZHANG Qian12CAO Minjie12
(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-endopeptidaseHaliotis discus hannaimetal ionscircular dichroism spectruminhibitors
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.

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更新日期/Last Update: 2019-06-17