|本期目录/Table of Contents|

[1]张玉梅,李鹏鹏,王晶晶,等.麻鸭钙非依赖性磷脂酶A2ε(iPLA2ε)的克隆和表达纯化[J].江苏农业科学,2018,46(17):170-175.
 Zhang Yumei,et al.Cloning,expression and purification of sheldrake calcium-independent phospholipase A2ε(iPLA2ε)[J].Jiangsu Agricultural Sciences,2018,46(17):170-175.
点击复制

麻鸭钙非依赖性磷脂酶A2ε(iPLA2ε)
的克隆和表达纯化
(PDF)
分享到:

《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第46卷
期数:
2018年第17期
页码:
170-175
栏目:
畜牧兽医与水产蚕桑
出版日期:
2018-09-05

文章信息/Info

Title:
Cloning,expression and purification of sheldrake calcium-independent phospholipase A2ε(iPLA2ε)
作者:
张玉梅12 李鹏鹏2 王晶晶12 王道营2 徐为民23
1.南京农业大学肉品加工与质量控制教育部重点实验室,江苏南京 210095; 2.江苏省农业科学院农产品加工研究所,江苏南京 210014;
3.江苏省肉类生产与加工质量安全控制协同创新中心,江苏南京 210095
Author(s):
Zhang Yumeiet al
关键词:
麻鸭iPLA2ε克隆原核表达纯化
Keywords:
-
分类号:
Q786;Q785
DOI:
-
文献标志码:
A
摘要:
肉品风味与肌内磷脂的水解密切相关,磷脂酶A2对肌内磷脂的水解起主导作用,其中钙非依赖性磷脂酶A2ε(iPLA2ε)是重要的一种。采用逆转录PCR技术,从麻鸭肌肉中克隆到iPLA2ε的编码基因。该基因开放阅读框长1 425 bp,编码474个氨基酸,分子量约为54 ku,理论等电点为7.86。生物信息学分析表明,iPLA2ε蛋白含有2个明显的跨膜区,为疏水性膜蛋白;iPLA2ε的氨基酸序列包含1个Patatin样磷脂酶结构域,该结构域中含有保守的丝氨酸-天冬氨酸催化水解二元体。采用大肠杆菌pCold1表达系统构建iPLA2ε基因的重组表达质粒,并在iPLA2ε基因的5′端连接大肠杆菌malE基因,该基因编码的麦芽糖结合蛋白能提高在原核中过量表达的真核融合蛋白的溶解性,最终在大肠杆菌中实现了iPLA2ε的可溶性表达。结果显示,利用蛋白C端的组氨酸标签亲和纯化法及凝胶过滤层析法获得了iPLA2ε的纯化蛋白,表明该研究为进一步研究肉品中iPLA2ε的功能奠定了工作基础。
Abstract:
-

参考文献/References:

[1]Liu L,Bartke N,van Daele H,et al. Higher efficacy of dietary DHA provided as a phospholipid than as a triglyceride for brain DHA accretion in neonatal piglets[J]. Journal of Lipid Research,2014,55(3):531-539.
[2]Ekeberg D,Storrustlkken L,Devle H M,et al. Lipid degradation and sensory characteristics of M. biceps femoris in dry cured hams from Duroc using three different processing methods[J]. International Journal of Food Science & Technology,2014,50(2):522-531.
[3]Wang D,Zhang M,Bian H,et al. Changes of phospholipase A2 and C activities during dry-cured duck processing and their relationship with intramuscular phospholipid degradation[J]. Food Chemistry,2014,145(7):997-1001.
[4]Gandemer G. Lipids in muscles and adipose tissues,changes during processing and sensory properties of meat products[J]. Meat Science,2002,62(3):309-321.
[5]Alasnier C,Gandemer G. Activities of phospholipases A and lysophospholipases in glycolytic and oxidative skeletal muscles in the rabbit[J]. Journal of the Science of Food and Agriculture,2000,80(6):698-704.
[6]Jenkins C M,Mancuso D J,Yan W,et al. Identification,cloning,expression,and purification of three novel human calcium-independent phospholipase A2 family members possessing triacylglycerol lipase and acylglycerol transacylase activities[J]. The Journal of Biological Chemistry,2004,279(47):48968-48975.
[7]Kim S J,Gershov D,Ma X J,et al. I-PLA2 activation during apoptosis promotes the exposure of membrane lysophosphatidylcholine leading to binding by natural immunoglobulin M antibodies and complement activation[J]. Journal of Experimental Medicine,2002,196(5):655-665.
[8]Xiao S,Zhang W A,Yang Y,et al. Changes of hormone-sensitive lipase (HSL),adipose tissue triglyceride lipase (ATGL) and free fatty acids in subcutaneous adipose tissues throughout the ripening process of dry-cured ham[J]. Food Chemistry,2010,121(1):191-195.
[9]Saarela J,Jung G,Hermann M,et al. The patatin-like lipase family in Gallus gallus[J]. BMC Genomics,2008,9(1):281.
[10]吴一凡,张双全,高秀玉,等. 乳糖诱导pET载体表达重组蛋白的研究[J]. 南京师大学报(自然科学版),2002,25(1):89-93.
[11]Kisel M A,Kuchuro S V,Litvinko N M. Influence of ethanol and phosphatidylethanol on the activity of pancreatic phospholipase A2[J]. Biochemistry,2001,66(2):168-172.
[12]Larsson P K,Claesson H E,Kennedy B P. Multiple splice variants of the human calcium-independent phospholipase A2 and their effect on enzyme activity[J]. The Journal of Biological Chemistry,1998,273(1):207-214.
[13]Liu X,Pan H,Yang G,et al. Cloning,expression and biochemical characterization of a basic-acidic hybrid phospholipase A2-Ⅱ from Agkistrodon halys Pallas[J]. Biochimica et Biophysica Acta,1999,1431(1):157-165.
[14]徐为民,卞欢,王道营,等. 鸭肉肌内磷脂酶解反应条件优化及其产物鉴定[J]. 江苏农业学报,2012,28(5):1130-1134.
[15]王道营. 肌内磷脂酶在传统板鸭加工中对磷脂的降解作用研究[D]. 南京:南京农业大学,2014.
[16]李海珍,李斌,张丽君,等. 竹叶青蛇毒磷脂酶A2的克隆与表达[C]//全国酶学学术讨论会.杭州,2011.
[17]Barroncasella E A,Kickler T S,Rogers O C,et al. Expression and purification of functional recombinant epitopes for the platelet antigens,PlA1 and PlA2[J]. Blood,1994,84(4):1157-1163.
[18]Premzl A,Kotnik V,Kveder R,et al. Urine analysis suggests association complement activation and apoptosis with membranous glomerulo patients nephritis[J]. Molecular Immunology,1999,36(4/5):306.
[19]Upreti G C,Hall E L,Koppens D,et al. Studies on the measurement of phospholipase A2 (PLA2) and PLA2 inhibitor activities in ram semen[J]. Animal Reproduction Science,1999,56(2):107-121.
[20]Terpe K. Overview of tag protein fusions:from molecular and biochemical fundamentals to commercial systems[J]. Applied Microbiology and Biotechnology,2003,60(5):523-533.

相似文献/References:

[1]王进,陈文强,邓百万,等.汉中麻鸭肠道定殖菌的生活状态和分布[J].江苏农业科学,2014,42(02):176.
 Wang Jin,et al.Living conditions and distribution of intestinal colonization bacteria in Hanzhong duck[J].Jiangsu Agricultural Sciences,2014,42(17):176.

备注/Memo

备注/Memo:
收稿日期:2017-04-11
基金项目:江苏省自然科学基金(编号:BK20161378)。
作者简介:张玉梅(1985—),女,河南永城人,硕士研究生,研究方向为肉品加工与质量控制研究。E-mail:2015808107@njau.edu.cn。
通信作者:徐为民,博士,研究员,主要从事肉品加工与质量控制研究,E-mail:weiminxu2002@aliyun.com;李鹏鹏,博士,主要从事肉品加工与质量控制研究,E-mail:lipp0514@gmail.com。
更新日期/Last Update: 2018-09-05