Screening and Identification of Protease-producing Bacillus amyloliquefaciens from Pig's Intestines and the Preliminary Study of Its Enzymatic Properties

A Bacillus strains,MY-6 strain which secreted high amount of protease was isolated from healthy pig's intestines,and it was identified as Bacillus amyloliquefaciens by the methods of morphological observation, biochemical tests,16S rDNA sequence amplification and comparison, biolog identification. In order to get an initial understanding of the enzymatic properties of its protease, the dynamic relationship between protease activity and bacterial life cycle, effects of pH,temperature,metal ions and inhibitors on protease activity,and the protease hydrolysis of different substrates were investigated. The results showed that,the protease synthesis of MY-6 strain was synchronized with its spore formation which began in the early stage of the exponential growth phase. The optimum conditions for the protease was 40℃ and pH 7.0,Ca²⁺ and Mn²⁺ had a intensive effects on the protease activity, PMSF could completely inhibit the activity of protease, indicating that the protease was a serine protease.The content of TCA soluble peptides for protein hydrolyzate fish protein, soybean protein, corn protein and casein was 0.83,0.67,0.58 and 0.38 μmol/g,respectively. In conclusion,Bacillus amyloliquefaciens MY-6 strain could be used as an initial selection of probiotic Bacillus to improve protein digestion.     

水解真空干燥法处理禽畜废弃物

提出了一种畜禽废弃物的处理方法——水解真空干燥法。水解真空干燥法是采用高温、高压进行消毒灭菌，通过抽真空来充分干燥物料的一种处理废弃物方法，对禽畜废弃物的病菌、病毒灭活较为彻底，处理过程安全可靠，能做到无害化处理；同时还可以最大程度保留废弃物中营养成分，可作为饲料添加剂。     

海带粒蜂蜜口服液的研制

以蜂蜜和海带为主要原料开发研制了一款口服液新产品.海带经过浸泡、清洗、切丝、熟化、打浆、打磨、过滤、调pH、酶解、灭酶、调配等工艺处理.通过正交试验得出酶解海带浆的最佳工艺参数为pH4.5、温度50℃、酶用量0.06％、酶解时间2h;通过评价正交试验样品,得出最佳配方为海带粒浆体50％、蜂蜜20％、海藻糖15％、柠檬酸0.2％、β-环糊精0.08％.     

以豆粕为原料制备反刍动物饲用肽的研究

本文通过正交试验,选用碱性微生物蛋白酶,研究得出酶解法制备大豆肽的最佳工艺参数:豆粕预处理条件为90℃水浴加热10min,酶解条件为底物浓度5%(W/V)、加酶量5万单位/g蛋白质、温度50℃、pH值10、酶解时间5.5h。蛋白质水解率达到25%,平均肽链长度为4.0。制得大豆肽粗蛋白质含量66.83%(DM)。并对制得大豆肽和原料豆粕的氨基酸含量进行分析。     

复种饲料油菜对麦茬耕层土壤酶活性的影响

采用大田试验方法,研究了不同种植密度与施氮量条件下,复种饲料油菜对麦茬耕层(0~15 cm)土壤酶活性的影响。结果表明,麦收后复种饲料油菜能显著或极显著地提高土壤过氧化氢酶(CAT)、酸性磷酸酶(APHA)、纤维素酶(CEL)和脲酶(URA)的活性,与休闲熟化地(CK)比较,最大提高幅度分别为32.15%,35.61%,78.66%和127.36%。土壤酶活性的变化与复种饲料油菜的播种量和施氮量有关,随播种量的增加土壤CAT、CEL和URA活性表现为逐渐递增态势,而土壤APHA表现为先增后减,播种量为11.5 kg/hm2时,APHA活性可达0.7888mg/g。在油菜生长的苗期,土壤APHA、CEL和URA随施氮量的增加而增加,土壤CAT则随施氮量的增加先增后降,呈单峰曲线;在生长后期(收获期),低氮量有利于土壤APHA活性的增加,中氮量有利于土壤CAT和CEL活性的增加,高氮量有利于URA活性的增加。     

豆粕酶解发酵物对仔猪生长性能、血清指标及肠道菌群结构的影响

本试验旨在研究豆粕酶解发酵物对仔猪生长性能、血清指标及肠道菌群结构的影响。选用144头49日龄健康的杜×长×大三元杂交育肥仔猪,随机分为3组,每组6个重复,每个重复8头猪(公母各占1/2)。阴性对照组饲喂无抗生素添加的基础饲粮,抗生素对照组饲喂基础饲粮+60 g/t吉他霉素,试验组饲喂试验饲粮(基础饲粮中10%豆粕被豆粕酶解发酵物替代)。试验期为42 d。结果表明:1)试验组仔猪的末重、平均日增重、平均日采食量分别比阴性对照组高5.06%、10.07%、5.03%(P0.05),腹泻率比阴性对照组低39.59%(P0.05),与抗生素对照组相比各指标差异均不显著(P0.05)。2)试验组仔猪血清碱性磷酸酶和谷丙转氨酶活性、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性均显著高于阴性对照组(P0.05),丙二醛(MDA)含量显著低于阴性对照组(P0.05);试验组和抗生素对照组仔猪血清指标差异不显著(P0.05)。3)试验组仔猪盲肠、大肠中乳酸杆菌数量显著高于阴性对照组和抗生素对照组(P0.05),十二指肠中总细菌数显著低于阴性对照组和抗生素对照组(P0.05)。综上,豆粕酶解发酵物能提高仔猪生长性能,增强机体抗氧化能力,利于乳酸杆菌在肠道内形成优势菌群,降低腹泻率,生长性能达到使用抗生素水平。     

糖蜜酒精废液处理过程中生成的微生物蛋白的最佳水解条件研究

通过实验选择复合蛋白酶水解糖蜜酒精废液处理过程中生成的微生物蛋白,并通过正交试验分析得到最佳水解条件为水解酶加入量5.5g/150mL,水解温度65℃,pH值5和水解时间5h。     

Serum bile acids: reference values in healthy dogs and comparison of two kit methods

Serum bile acid (SBA) reference intervals were established by use of a radioimmunoassay method for fasting dogs to be 0.2 to 4.3 micro mol/L (n = 60) and for 2 hour postprandial samples to be 0.6 to 24.2 micro mol/L (n = 37). The SBA reference intervals estimated using an enzymatic method were 0 to 8.6 micro mol/L for fasting (n = 26) and 0 to 29.8 micro mol/L for 2 hour postprandial samples (n = 36). The correlation between the two methods including samples from healthy dogs and clinical cases is good (n = 128, r = 0.82, p < 0.0001). The radioimmunoassay method is linear to 50 micro mol/L and the enzymatic method is linear to 100 micro mol/L, thus both methods require serum dilutions to be made in many cases of primary liver disease. The enzymatic method is less expensive and more convenient for use in a clinical laboratory but requires a greater sample volume (400 micro I) than the RIA method (50 micro I). Both methods have adequate precision and accuracy to be useful as diagnostic tests of liver function in dogs.     

鹿血清蛋白抗氧化肽体外抗氧化性及其机理研究

在碱性蛋白酶最适条件下水解鹿血清蛋白,水解时间为0~5 h,鹿血清蛋白抗氧化肽的水解度(DH)随着水解时间增加而增加(P0.05),鹿血清蛋白抗氧化肽还原能力、清除自由基的作用和Cu2+螯合能力随着水解时间的增加而显著提高(P0.05),鹿血清蛋白抗氧化肽能有效抑制硫代巴比妥酸反应物质(TBARS)生成(P0.05)。结果表明,鹿血清蛋白抗氧化肽的抗氧化活性取决水解时间,鹿血清蛋白抗氧化肽是有效的抗氧化剂。