海参组织蛋白酶K的酶学性质及其对海参自溶的影响
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- 英文论文题名：Cathepsin K of sea cucumber stichopus japonicus:properties and effect on autolysis
- 论文作者：季晓彤 ; 王玲 ; 王婷 ; 薛鹏 ; 年益莹 ; 孙黎明
- 英文论文作者：Ji Xiaotong ; Wang Ling ; Wang Ting ; Xue Peng ; Nian Yiying ; Sun Liming ; School of Food Science and Technology ; Dalian Polytechnic University ; National Engineering Research Center of Seafood
- 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;
- 论文关键词：海参 ; 组织蛋白酶K ; 自溶
- 英文论文关键词：sea cucumber ; cathepsin K ; autolyze
- 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：TS254.1
- 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：111k
- 原文格式：O
- 会议级别：全国
摘要
本文对海参体壁组织蛋白酶K(CatK)的酶学性质及其对海参自溶的影响进行了初步探讨。采用特异性荧光底物法(以Z-Gly-Pro-Arg-MCA为底物)考察了CatK的基本酶学性质。结果表明CatK的最适pH为5.0,最适温度为50℃,在20-40℃之间稳定性较好。Ca~(2+)、Mg~(2+)、Mn~(2+)、Zn~(2+)、Fe~(2+)、Cu~(2+)均对该酶产生一定抑制作用,后四种离子对CatK的抑制率均超过87%。CA-074、Z-Leu-Leu-Leu-H(ZLLL)、碘乙酸、E-64、抗痛素、PMSF对CatK的抑制率均在85%以上,1,10-菲啰啉和EDTA对该酶的抑制率分别为16%和39.7%,DTT和L-Cys可将该酶活力分别提高至127.46%和238.3%。在海参肉中加入抑制剂CA-074和ZLLL,于25℃孵育使海参肉自溶,用SDSPAGE研究CatK对海参蛋白降解的影响,结果显示ZLLL能够在一定程度上抑制海参蛋白的降解。上述结果表明海参CatK是一种巯基氨基酸含量较高的半胱氨酸蛋白酶,具有一定的金属离子依赖性,但又易被多种金属离子抑制;CatK有可能直接参与海参自溶过程中蛋白质的降解。
Properties of cathepsin K(CatK) in sea cucumber body wall and its influence on autolysiswere studied.Specificity substrate fluorescence method(using Z-Gly-Pro-Arg-MCA as substrate) was used to investigate someproperties of CatK.Results showed that a maximum activity was observed at pH5.0 and 50 t,the activity kept stable at 20-40℃.CatK activity was inhibited by Ca~(~(2+)),Mg~(2+),Mn~(2+),Zn~(2+),Fe~(2+),Fe~(3+),Cu~(2+),more than 87%of the activity was the inhibited by the latter four ions.CA-074,Z-Leu-Leu-Leu-H(ZLLL),iodoacetate,E-64,antipain,PMSF could inhibit CatK activity with an inhibitory rate of over 85%.1,10-phenanthroline and EDTA showed inhibition rateof 16%and 39.7%,respectively.Compared with the control group,DTT and L-Cys CatKcould promotethe activity up to percentages of 127.46%and 238.3%,respectively.CA-074 and ZLLL were then added into sea cucumber meat,the mixture was incubated at 25℃ for the meat todevelop autolysis.The effect of CA-074 and ZLLLon protein degradation of sea cucumber was examined by SDS-PAGE.Result indicated thatZLLLcould inhibit the degradation of protein to a certain degree.Above results indicated that the CatK in sea cucumber was a typical cysteine protease,ion dependent but also sensitive tomany metal ions.It might directly participate in the protein degradation during autolysis of sea cucumber.
Properties of cathepsin K(CatK) in sea cucumber body wall and its influence on autolysiswere studied.Specificity substrate fluorescence method(using Z-Gly-Pro-Arg-MCA as substrate) was used to investigate someproperties of CatK.Results showed that a maximum activity was observed at pH5.0 and 50 t,the activity kept stable at 20-40℃.CatK activity was inhibited by Ca~(~(2+)),Mg~(2+),Mn~(2+),Zn~(2+),Fe~(2+),Fe~(3+),Cu~(2+),more than 87%of the activity was the inhibited by the latter four ions.CA-074,Z-Leu-Leu-Leu-H(ZLLL),iodoacetate,E-64,antipain,PMSF could inhibit CatK activity with an inhibitory rate of over 85%.1,10-phenanthroline and EDTA showed inhibition rateof 16%and 39.7%,respectively.Compared with the control group,DTT and L-Cys CatKcould promotethe activity up to percentages of 127.46%and 238.3%,respectively.CA-074 and ZLLL were then added into sea cucumber meat,the mixture was incubated at 25℃ for the meat todevelop autolysis.The effect of CA-074 and ZLLLon protein degradation of sea cucumber was examined by SDS-PAGE.Result indicated thatZLLLcould inhibit the degradation of protein to a certain degree.Above results indicated that the CatK in sea cucumber was a typical cysteine protease,ion dependent but also sensitive tomany metal ions.It might directly participate in the protein degradation during autolysis of sea cucumber.
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