摘要
肌纤维类型是决定肉及肉制品品质特性的内因,肌球蛋白亚型决定了肌纤维类型。为突破不同肌球蛋白亚型难以从混合肌纤维中分离纯化的难点,以牛咬肌(100%的Ⅰ型肌纤维)和牛躯干皮肌(93%的Ⅱ型肌纤维)为原料,确立一种可同时适用于两种肌球蛋白亚型的提取及纯化方法。从已报道的4种不同物种源肌球蛋白提取方法中筛选出较适宜的一种,进而对其组织破碎程度、肌浆蛋白去除效果、硫酸铵分级沉淀效果进行优化,通过十二烷基硫酸钠聚丙烯酰胺电泳和纳升液相色谱-四极杆静电场轨道阱高分辨质谱联用技术分析提取物的蛋白质组成及肌球蛋白亚型纯度。结果表明:分别从牛咬肌和躯干皮肌中获得纯度高达95%的Ⅰ型肌球蛋白和93%的Ⅱ型肌球蛋白。本研究确立的方法可为促进肉品品质研究深入到肌球蛋白亚型水平提供技术可行性,为推动肌球蛋白亚型与肉品品质特性关系的研究奠定基础。
Muscle fiber type is the internal factor that determines the quality characteristics of meat and meat products,and myosin subtype determines muscle fiber type.In order to overcome the difficulty of separating and purifying different myosin subtypes from mixed muscle fibers,this study took bovine masseter(100%typeⅠmuscle fibers)and bovine cutaneous trunci(93%typeⅡmuscle fibers)as objects to establish a method for extraction and purification of both myosin subtypes.Among the four reported extraction methods for myosin from different species,a more suitable method was selected,and then the degree of tissue fragmentation,removal efficiency of sarcoplasmic proteins,and fractional precipitation effects of ammonium sulfate were optimized.The protein composition and the purity of myosin subtypes were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and nano-liquid chromatography coupled with quadrupole orbitrap high-resolution mass spectrometry.The results showed that myosin with a purity of up to 95%typeⅠand 93%typeⅡwas obtained from bovine masseter muscle and cutancous trunci,respectively.The method established in this study provides technical feasibility for promoting the research of meat quality to the level of myosin subtypes and lays a foundation for promoting the study of the relationship between myosin subtypes and meat quality characteristics.
作者
高宏敏
宋尚新
刘安凤
秦怡
王纪
高雨琳
周峰
曹崇江
GAO Hongmin;SONG Shangxin;LIU Anfeng;QIN Yi;WANG Ji;GAO Yulin;ZHOU Feng;CAO Chongjiang(College of Engineering,China Pharmaceutical University,Nanjing 211198,China;School of Food Science,Nanjing Xiaozhuang University,Nanjing 211171,China)
出处
《保鲜与加工》
CAS
北大核心
2024年第10期58-66,共9页
Storage and Process
基金
国家重点研发计划项目(2022YFF1100600)
国家自然科学青年基金项目(32202099)
江苏省高等学校基础科学(自然科学)面上项目(21KJB550008)。
关键词
肌纤维类型
肌球蛋白亚型
咬肌
躯干皮肌
提取
纯化
muscle fiber type
myosin subtype
masseter
cutancous trunci
extraction
purification
