南美白对虾丝氨酸蛋白酶抑制剂的筛选及其对自溶的控制研究
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- 英文论文题名：Inhibitors against SP in southern white shrimp litopenaeus vannamei:screening and effect on autolysis
- 论文作者：季晓彤 ; 王玲 ; 王婷 ; 崔钰婷 ; 年益莹 ; 薛鹏 ; 孙黎明
- 英文论文作者：Ji Xiaotong ; Wang Ling ; Wang Ting ; Cui Yuting ; Nian Yiying ; Xue Peng ; 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：南美白对虾 ; 丝氨酸蛋白酶 ; 自溶 ; 抑制剂
- 英文论文关键词：southern white shrimp ; serine protease ; autolysis ; inhibitor
- 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：106k
- 原文格式：O
- 会议级别：全国
摘要
本文基于南美白对虾体内丝氨酸蛋白酶(SP)抑制剂的筛选,为控制对虾自溶提供有效方法。首先采用特异性荧光底物法(Boc-Phe-Ser-Arg-MCA),对SP的酶学性质进行研究,结果表明,其最适温度为50℃,在20-50℃之间稳定性较好;最适pH为9.0,在pH7.0-9.0之间稳定性较好;Zn~(2+)可强烈抑制SP的活性;马铃薯、大蒜、黄豆水提物和新鲜蛋清对SP的活性均有显著抑制作用;山梨酸钾、柠檬酸、EDTA也能够在一定程度上抑制SP的活性。利用SDS-PAGE考察了南美白对虾虾糜的自溶情况以及上述抑制剂对其自溶的影响。25℃孵育2h,虾糜发生明显自溶,肌球蛋白重链和肌动蛋白几乎全部降解。马铃薯水提物、柠檬酸可显著抑制虾糜自溶,山梨酸钾、蛋清对虾糜自溶也有一定抑制作用,而黄豆水提物、Zn~(2+)、大蒜水提物、EDTA对虾肉自溶没有明显的抑制效果。结果提示,丝氨酸蛋白酶抑制剂的筛选,是一种快速筛选南美白对虾自溶控制剂的有效方法;另外,对虾体内除了SP发挥自溶作用,很可能存在其他类型参与自溶的蛋白酶。
Basing on inhibitors screening of the serine protease(SP) from southern white shrimps,effective method was discussed to control autolysis of shrimp.Firstly,SP's enzymatic property of Litopenaeus vannamei was studied by specificitysubstrate fluorescence method(Boc-Phe-Ser-Arg-MCA).Results showed that a maximum activity was observed at pH9.0 and50℃,the activity kept stableat 20-60℃ and.pH 7.0-9.0.The SP's activity was significantly inhibited by Zn~(2+) ions.The aqueous extract of potatoes,garlic,yellow bean and fresh egg white had certain inhibition effect on SP's activity.Potassium sorbate,citric acid,Ethylene Diamine Tetraacetic Acid(EDTA) also inhibited the SP's activity to some extent.Autolysis degree of shrimp mince and the effect of the above factors which could restrain SP's activivy were examined by SDS-PAGE.Result indicated that the serious autolysis occurred after 2h's incubation at 25℃,both myosin heavy chain and actin chain degraded obviously.Both potatoes aqueous extract and citric acid could inhibit autolysis ofshrimp meat obviously.Also,potassium sorbate and egg white also demonstrated a weak inhibitory effect.However,soybean aqueous extract,Zn~(2+),garlic aqueous extract and EDTA had no evident effect on autolysis.Above all,it might be an effective and quick method to find method to control autolysis of shrimp by screening inhibitors of SPs.In addition,there are probably other typesof protease participating in autolysis besides SPs
Basing on inhibitors screening of the serine protease(SP) from southern white shrimps,effective method was discussed to control autolysis of shrimp.Firstly,SP's enzymatic property of Litopenaeus vannamei was studied by specificitysubstrate fluorescence method(Boc-Phe-Ser-Arg-MCA).Results showed that a maximum activity was observed at pH9.0 and50℃,the activity kept stableat 20-60℃ and.pH 7.0-9.0.The SP's activity was significantly inhibited by Zn~(2+) ions.The aqueous extract of potatoes,garlic,yellow bean and fresh egg white had certain inhibition effect on SP's activity.Potassium sorbate,citric acid,Ethylene Diamine Tetraacetic Acid(EDTA) also inhibited the SP's activity to some extent.Autolysis degree of shrimp mince and the effect of the above factors which could restrain SP's activivy were examined by SDS-PAGE.Result indicated that the serious autolysis occurred after 2h's incubation at 25℃,both myosin heavy chain and actin chain degraded obviously.Both potatoes aqueous extract and citric acid could inhibit autolysis ofshrimp meat obviously.Also,potassium sorbate and egg white also demonstrated a weak inhibitory effect.However,soybean aqueous extract,Zn~(2+),garlic aqueous extract and EDTA had no evident effect on autolysis.Above all,it might be an effective and quick method to find method to control autolysis of shrimp by screening inhibitors of SPs.In addition,there are probably other typesof protease participating in autolysis besides SPs
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