菲律宾蛤仔足肌肌肉组织学特性及蛋白分离机制  

The Muscle Histology and Protein Separation Mechanism on Foot Muscle of Ruditapes philippinarum

作　　者：张龙 刘俊荣 田元勇 郑尧 刘慧慧 宋扬 姜梦云 刘金洋 Zhang Long;Liu Junrong;Tian Yuanyong;Zheng Yao;Liu Huihui;Song Yang;Jiang Mengyun;Liu Jinyang(College of Food Science and Engineering,Dalian Ocean University,Dalian 116023,Liaoning)

机构地区：[1]大连海洋大学食品科学与工程学院,辽宁大连116023

出　　处：《中国食品学报》2018年第7期90-98,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金面上项目(041314030)

摘　　要：为探索菲律宾蛤仔平滑肌中蛋白质的分离特性,以足肌为研究对象,从肌肉组织学出发,以肌球和副肌球蛋白为重点,对肌肉组织形态、肌原纤维蛋白(Mf)分布特性以及肌球和副肌球蛋白分离特性等进行分析。对足肌进行组织学观察,分析肌纤维的形态与分布;利用SDS-PAGE电泳对Mf分布特性进行定性和定量分析;通过调节KCl、ATP、硫酸铵(A.S.)浓度,以ATPase活性为蛋白质变性指示参数,分析各肌原纤维蛋白组分的解离与析出规律,并确定肌球和副肌球蛋白的分离纯化条件。研究结果表明:(1)肌纤维呈三维网状分布,肌肉层可分为横向肌、纵向肌和环向肌;Mf组分中存在平滑肌标志性蛋白Myorod,其中肌球蛋白和副肌球蛋白分别占15%和30%左右。(2)Mf的溶解性依赖于KCl和ATP浓度,Ca2+-ATPase活性在低离子强度下较高,确定KCl浓度为0.1 mol/L;肌球蛋白最佳解离和析出条件分别为:5~10 mmol/L ATP缓冲液梯度溶解,45%~60%饱和A.S.分级沉淀;副肌球蛋白最佳解离和析出条件分别为:10~20 mmol/L ATP缓冲液梯度溶解,30%~40%饱和A.S.分级沉淀。In this study, the foot muscle morphology, myofibrillar protein（Mf） distribution and the separation profiles of myosin and paramyosin were analyzed with emphasis on myosin and paramyosin, regarding the muscle histology as the starting point, for exploring the protein separation profiles on smooth muscle of Ruditapes philippinarum. Firstly,the morphology and distribution of foot muscle fibers were explored through histological observation; Secondly, The qualitative and quantitative analysis on Mf were performed by SDS-PAGE; Thirdly, the regulation of dissociation and precipitation in Mf was investigated, then the separation and purification conditions of myosin and paramyosin were determined through adjusting the concentration of potassium chloride, adenosine triphosphate and ammonium sulfate with regarding the ATPase as indicator parameters of protein denaturation. The results were indicated that（1） The muscle layers arranged in three dimensional network could be divided into transverse, longitudinal and circular fibers. Myorod as the symbolic protein of smooth muscle is present in Mf, the proportion of myosin and paramyosin in Mf is about 15% and 30%, respectively.（2） The solubility of Mf is depended on ionic strength and ATP. The higher values of Ca2＋-ATPase activity are present in the low ionic strength then 0.1 mol/L is determined. The optimal conditions of dissociation and precipitation of myosin is 5-10 mmol/L ATP buffer solution to dissolve and 45%-60% saturation A.S. to precipitate, as well as,paramyosin is 10-20 mmol/L ATP buffer solution to dissolve and 30%-40% saturation A.S. to precipitate, respectively.

关 键 词：菲律宾蛤仔 平滑肌 肌球蛋白 副肌球蛋白 分离机制 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]