复合酶制剂体外预消化仔猪饲粮最佳条件的筛选以及对其品质的影响  

Screening of Optimal Conditions for Piglet Diet Predigested by Compound Enzyme Preparation in Vitro and Effects on Its Quality

作　　者：杨阳 王悦 武一帆 刘思均 赵燕[1] 李步高[1] 曹果清[1] 郭晓红[1] YANG Yang;WANG Yue;WU Yifan;LIU Sijun;ZHAO Yan;LI Bugao;CAO Guoqing;GUO Xiaohong(College of Animal Science,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学动物科学学院,太谷030801

出　　处：《动物营养学报》2022年第5期3329-3338,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：三晋学者支持计划专项经费资助(2016,2017);山西省“1331”工程(2021);山西省青年基金(201901D211369);山西省优秀博士奖励(SXYBKY2019030);山西农业大学科技创新(2020BQ01)。

摘　　要：本研究旨在筛选出体外预消化复合酶制剂的最佳作用条件,比较复合酶预消化前后饲粮的差异,为无抗饲粮的研发提供参考。试验选用蛋白酶、淀粉酶、非淀粉多糖(NSP)类酶(纤维素酶、果胶酶、木聚糖酶、β-葡聚糖酶和α-半乳糖苷酶)、葡萄糖氧化酶(GOD)、阿魏酸酯酶(FAE)和阿拉伯呋喃糖苷酶(AF)10种酶,组成复合酶制剂,根据酶的理化特性,探索酶的作用温度和时间对预消化仔猪饲粮的影响,应用扫描电镜和激光共聚焦拍照技术评定酶解前后饲粮的结构变化,并通过细菌生长曲线观察酶解预消化饲粮对肠道有害菌的影响。结果表明:1)与未添加酶相比,添加蛋白酶,在pH=3的缓冲液中预消化,50℃作用0.5 h时,氨基酸和小肽分子含量显著升高(P<0.05);添加淀粉酶和NSP酶,在pH=5的缓冲液中反应,预消化1.0 h后,二硝基水杨酸(DNS)法检测得到50℃时还原糖含量极显著升高(P<0.01),酶联免疫吸附试验(ELISA)检测发现50℃时部分抗营养因子含量也显著降低(P<0.05)。2)本试验分析木聚糖主链酶与其支链酶的协同作用,结果显示先添加AF和FAE,再添加木聚糖酶后酶解饲粮,还原糖的释放量比单独添加木聚糖酶提高了79.07%(P<0.01)。此外,再添加GOD,产生的葡萄糖酸含量为1.32μg/mL,并且降低了饲粮的pH。3)扫描电镜和罗丹明B染色结合激光共聚焦拍照检测发现,经过复合酶制剂预消化,饲粮的细胞壁损伤严重,完整结构遭到破坏。4)本试验选用从腹泻猪肠道中分离出的大肠杆菌和沙门氏菌来检验饲粮酶解产物的抑菌作用,结果表明酶解预消化后可以抑制大肠杆菌和沙门氏菌的生长。综上所述,酶制剂的最佳组合条件为:首先在pH=3、50℃下,加入蛋白酶,作用时间为0.5 h;接着上调pH为5,加入AF、FAE和GOD,40℃作用1.0 h;最后加入淀粉酶和NSP酶群50℃作用1.0 h。酶解后的饲粮,其可利用的营养物质含量提高,抗营养因子�The purpose of this study was to screen out the optimal conditions of compound enzyme preparation for in vitro predigestion and compare the difference of diets before and after compound enzyme predigestion,so as to provide reference for the development of non-resistant diet. Ten enzymes including protease,amylase,non-starch polysaccharide(NSP)enzymes(cellulase,pectinase,xylanase, β-glucanase andα-galactosidase),glucose oxidase(GOD),ferulic acid esterase(FAE)and arabinofuranosidase(AF)were selected to form a compound enzyme preparation. According to the physical and chemical properties of enzymes,the effects of enzyme action temperature and time on predigested diet for piglets were investigated,the structural changes of diets before and after enzymatic hydrolysis were evaluated by scanning electron microscopy and laser confocal photography,and the effects of predigested diet on intestinal harmful bacteria were observed by bacterial growth curve. The results showed as follows:1)compared with no enzyme adding,the contents of amino acids and small peptides were significantly increased by adding protease in buffer solution with pH = 3 and treated at 50 ℃ for 0.5 h(P<0.05);when amylase and NSP enzyme were added and reacted in buffer solution with pH = 5 and after pre-digestion for 1.0 h,the dinitrosalicylic acid(DNS)method showed that reducing sugar content was extremely significantly increased at 50 ℃(P<0.01),and enzyme-linked immunosorbent assay(ELISA)showed that the contents of some anti-nutritional factors were significantly decreased at 50 ℃(P<0.05). 2)The synergistic effect of xylanase and its branched enzyme was analyzed in this experiment.The results showed that the reducing sugar release content in diets adding AF and FAE first and then xylanase was increased by 79.07%(P<0.01)than that adding xylanase alone. In addition,adding GOD resulted in1.32μg/mL gluconic acid,and decreased the pH of the diet. 3)Scanning electron microscopy and rhodamine B staining combined with laser confocal photography showed

关 键 词：复合酶制剂 饲粮预消化 阿魏酸酯酶 阿拉伯呋喃糖苷酶 葡萄糖氧化酶 

分 类 号：S816[农业科学—饲料科学]