桂牧一号杂交象草在人工瘤胃培养液中蛋白质及氨基酸的消化规律研究  

Study on digestibility of protein and amino acid of Pennisetum purpureum cv.guimu-1 in artificial rumen culture medium

作　　者：杨膺白[1] 李菁华 赵雅丽 叶添梅 蔡小艳 于凯 YANG Yingbai;LI Jinghua;ZHAO Yali;YE Tianmei;CAI Xiaoyan;YU Kai(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;The Sericultural Research Institute of Hunan Province,Changsha 410127,China;College of Agriculture,Ningxia University,Yinchuan 750021,China)

机构地区：[1]广西大学动物科学技术学院,南宁530004 [2]湖南省蚕桑科学研究所,长沙410127 [3]宁夏大学农学院,银川750021

出　　处：《黑龙江畜牧兽医》2021年第22期103-108,152,153,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：广西科技基地和人才专项(桂科AD17129043)。

摘　　要：为了研究桂牧一号杂交象草在人工瘤胃培养液中蛋白质和氨基酸的消化规律,筛选消化受阻蛋白并分析其功能,试验将3只黑山羊成年羯羊的瘤胃液与人工唾液以1∶2的体积比混合,配制成人工瘤胃培养液,采用产气法消化10 g桂牧一号杂交象草,测定桂牧一号杂交象草原料及在人工瘤胃培养液中不同消化时间点(12,24,48小时)的干物质、蛋白质(粗蛋白、真蛋白、可溶性粗蛋白)相对含量,计算其消化率;采用定量蛋白组学筛选不同消化时间点的消化受阻蛋白,并对消化12 h的消化受阻蛋白进行亚细胞定位、基因本体(GO)注释和酶活性注释;测定不同消化时间点桂牧一号杂交象草样品中氨基酸含量及消化率。结果表明:桂牧一号杂交象草样品随消化时间的延长,粗蛋白和真蛋白的消化率逐渐升高,消化24小时时的可溶性粗蛋白消化率低于12小时时;与0小时对比,消化12小时时鉴定到66个消化受阻蛋白,消化24小时时鉴定到105个消化受阻蛋白,消化48小时时鉴定到112个消化受阻蛋白;消化12,24,48小时时的消化受阻蛋白都主要定位于叶绿体中;消化12小时时的消化受阻蛋白主要分布在RNA代谢过程、细胞组分发生、大分子复合物亚基组成等生物学过程,类囊体部分、类囊体、叶绿体类囊体、质体类囊体等细胞组分,结合、杂环化合物结合、有机环状化合物结合等分子功能;消化12小时时的消化受阻蛋白具有水解酶功能的蛋白数量最多,其次是转移酶、氧化还原酶等;桂牧一号杂交象草氨基酸含量整体呈0~24 h上升、24~48 h降低的趋势,消化12小时时,丙氨酸、缬氨酸、异亮氨酸、亮氨酸、赖氨酸、组氨酸、精氨酸的消化率较低(<10%),其中组氨酸消化率最低(0.41%)。说明在人工瘤胃培养液条件下,桂牧一号杂交象草的蛋白质和氨基酸的消化存在阻滞现象。This experiment was conducted to investigate the digestibility of protein and amino acid of Pennisetum purpureum cv.Guimu-1 in artificial rumen,screen digestive block-related proteins(DRPs)and analyze its function.The rumen fluid of three adult Capricorn black goats was mixed with artificial saliva at the volume ratio of 1∶2 to prepare artificial rumen culture medium,10 g Pennisetum purpureum cv.Guimu-1 was digested by gas production method procedure.The relative contents of dry matter and protein(crude protein,true protein and soluble crude protein)of Pennisetum purpureum cv.Guimu-1 at different digestion time points(12,24 and 48 h)in artificial rumen culture medium were measured,and its digestibility was calculated.Quantitative proteomics was used to screen the DRPs at different digestion time points.Subcellular localization,Gene Ontology(GO)annotation and enzyme activity annotation were performed for the DRPs digested at 12 h.The content and digestibility of amino acid of Pennisetum purpureum cv.Guimu-1 were tested.The results showed that the digestibility of crude protein and true protein of pennisetum purpureum cv.guimu-1 increased with digestion time,while the digestibility of soluble crude protein at 24 h was lower than 12 h.66 DRPs were identified at 12 h,105 DRPs at 24 h and 112 DRPs at 48 h compared to 0 h.At 12,24 and 48 hours of digestion,the DRPs were mainly located in chloroplasts.The DRPs at 12 h were mainly involved in biological processes including RNA metabolism process,cellular component biogenesis,subunit composition of macromolecular complexes;and distributed in cellular components including thylakoid part,thylakoid,chloroplast thylakoid and plastid thylakoid;and related to molecular functions including binding,heterocyclic compounds binding,organic cyclic compound binding.In addition,DRPs at 12 h were most involved in hydrolase function,followed by transferase,oxidoreductase and so on.Moreover,the amino acid content of Pennisetum purpureum cv.Guimu-1 increased at 0-24 h and decreased at 24

关 键 词：桂牧一号杂交象草 消化受阻蛋白 蛋白质组学 人工瘤胃液 氨基酸 

分 类 号：S815[农业科学—畜牧学]