内蒙古地区传统发酵酸马奶不同发酵时期乳清蛋白SDS-PAGE分析  

SDS-PAGE Analysis of Whey Protein at Different Fermentation Stages of Traditional Koumiss from Inner Mongolia

作　　者：红梅[1] 包艳青 芒来[3] 布仁其其格[1] Hongmei;BAO Yanqing;DUGARJAVIIN Manglai;Burenqiqige(Inner Mongolia Medical University,Hohhot 010110,China;Agricultural and Animal Husbandry Technology Extension Center of Hinggan League of Inner Mongolia,Ulanhot 137400,China;Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古医科大学,内蒙古呼和浩特010110 [2]内蒙古兴安盟农牧技术推广中心,内蒙古乌兰浩特137400 [3]内蒙古农业大学,内蒙古呼和浩特010018

出　　处：《畜牧与饲料科学》2023年第5期83-92,共10页Animal Husbandry and Feed Science

基　　金：内蒙古医科大学博士启动基金(YKD2018BSJJ008);内蒙古医科大学大学生创新创业训练计划项目(202010132050);内蒙古医科大学大学生创新创业培育项目(SWYT2020027)。

摘　　要：[目的]探究内蒙古地区传统酸马奶发酵过程中乳清蛋白的变化情况。[方法]以从内蒙古锡林郭勒盟镶黄旗采集的马奶为原料制备传统发酵酸马奶,发酵温度为(22±2)℃,人工捣拌频率5次/d,每次300下;每隔12 h采集1次发酵0~96 h的酸马奶样品;离心提取酸马奶乳清蛋白样品,利用条件优化后的SDS-PAGE电泳技术分析酸马奶乳清蛋白在发酵过程中变化情况。[结果]优化后的SDS-PAGE电泳分析条件为:分离胶浓度12%,浓缩胶浓度5%,分离胶电泳电压180 V、电泳时间300 min,上样浓度0.5μg/μL,上样体积16μL,改良考马斯亮蓝法染色、脱色各12 h。在发酵0~12 h内,酸马奶乳清蛋白含量增加,在12 h时达到最高值(5.12μg/μL),之后随发酵时间延长呈下降趋势,发酵96 h时降低至3.92μg/μL。SDS-PAGE电泳图谱分析显示,在分子量10~70 kDa区间内共获得乳清蛋白有效条带13个。在发酵过程中,β-乳球蛋白、溶酶菌C和α-乳白蛋白3种蛋白质含量呈现不同程度的波动,发酵72 h时三者含量之和最高,占乳清蛋白总含量的85.73%,并且在发酵过程中始终保持较高水平,是传统发酵酸马奶发酵96 h内乳清蛋白的主要组分,其中,β-乳球蛋白含量最高,平均占乳清蛋白总含量的38.92%。分子量在20~40 kDa的酪蛋白含量在发酵24 h之后开始下降,至60 h时条带消失。分子量为10.33 kDa的蛋白质在发酵12 h内被消耗,60 h时再次出现且含量随发酵时间延长逐渐增加,在84 h时达到峰值,占乳清蛋白总含量的9.84%。分子量为49.80 kDa的蛋白质在发酵72 h之后开始出现,含量在96 h时达到乳清蛋白总含量的4.11%。与其他乳清蛋白组分相比,β-乳球蛋白和免疫球蛋白含量在发酵过程中变化较小。[结论]内蒙古地区传统发酵酸马奶在发酵96 h内,分子量为10~70 kDa的乳清蛋白含量表现出有规律的消长,主要被消化分解的蛋白质是酪蛋白。不同发酵阶段酸马奶乳清蛋白组成�[Objective]This study was conducted to characterize the changes in whey protein profile during the fermentation process of traditional koumiss from Inner Mongolia.[Method]Traditional koumiss was prepared using mare milk collected from Xianghuang Banner,Xilin Gol League,Inner Mongolia,with fermentation temperature at(22±2)℃,and manual mixing 5 times per day,300 beats each time.The koumiss samples were taken every 12 h during the 0-96 h fermentation process,and whey protein was collected by centrifugation.Variations in Koumiss whey protein profile during the fermentation process were analyzed by the optimized SDS-PAGE.[Result]The optimized SDS-PAGE conditions were as follows:the concentrations of separation gel and spacer gel were 12%and 5%,respectively;the electrophoresis voltage for separation gel was 180 V,and the electrophoresis time was 300 min;the sample loading amount and concentration were 16μL and 0.5μg/μL samples,respectively;the gel was stained with modified Coomassie brilliant blue for 12 h and then decolorized for 12 h.Within 0-12 h of fermentation,the koumiss whey protein contents increased and peaked(5.12μg/μL)at 12 h,and then fell with the extension of fermentation time,decreasing to 3.92μg/μL at 96 h of fermentation.SDS-PAGE analysis showed that a total of 13 effective bands of whey protein within the molecular weight range of 10-70 kDa were observed.During the fermentation process,the contents of 3 types of whey protein,β-lactoglobulin,lysozyme C,andα-lactalbumin,fluctuated to varying degrees.The sum of their contents peaked at 72 h of fermentation,accounting for 85.73%of the total whey protein contents and maintained high levels during the fermentation process.It was indicated that they were the main components of whey protein in traditional koumiss within the fermentation process of 96 h.Among them,the content ofβ-lactoglobulin was the highest,accounting for an average of 38.92%of the total whey protein contents.The content of casein with the molecular weight range of 20-40 kDa bega

关 键 词：酸马奶 传统发酵 乳清蛋白 SDS-PAGE凝胶电泳 内蒙古 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程] TS252.54[轻工技术与工程—食品科学与工程] TS252.7