黄淮白山羊瘤胃微生物Fosmid文库的构建与分析

采用CTAB法和蛋白酶K法提取黄淮白山羊瘤胃微生物总DNA,酶切获得DNA片段,大小为23～50 kb,以E.coli EPI300为宿主细胞,构建瘤胃微生物宏基因Fosmid文库。研究发现该文库共获得克隆28 800个,插入片段大小约35 kb,空载率小于2%,容量1 008 Mb。通过羧甲基纤维素酶和木聚糖酶(Xylanase)活性筛选,分别获得具有两种酶活性的阳性克隆60和32个,木聚糖酶和羧甲基纤维素酶(CMC,Carboxymethyl cellulase)共同作用阳性克隆15个。通过已测序MF3木聚糖酶阳性克隆酶学活性分析,该木聚糖酶在最适应pH 4.5, 40℃条件下,酶活力为79.287μ·mL~(-1)。为进一步研究黄淮白山羊瘤胃内纤维素酶(Cellulase)基因奠定基础。     

瘤胃厌氧真菌和细菌对木质素含量不同底物的发酵特性

利用体外发酵法研究了瘤胃厌氧真菌和细菌对木质素含量不同底物(黑麦草叶、黑麦草茎、稻草秸、花生壳,木质素含量依次上升)的发酵特性.结果表明:瘤胃厌氧真菌体外发酵不同底物时的96 h累积产气量、底物干物质消失率和纤维素消失率差异均不显著,瘤胃细菌体外发酵时的干物质消失率和纤维素消失率均随底物木质素含量的上升而显著下降(P<0.05).此外,各真菌培养组96 h发酵液中,稻草秸组的木聚糖酶酶活力显著高于黑麦草叶和黑麦草茎(P<0.05),发酵相同底物时,真菌产生的木聚糖酶酶活力是羧甲基纤维素酶酶活力的23-65倍.各细菌培养组96 h发酵液中,黑麦草叶和黑麦草茎的木聚糖酶和羧甲基纤维素酶酶活力显著高于稻草秸(P<0.05).     

特异腐质酶Humicola insolens Y1耐热型木聚糖酶基因的克隆及酶学性质分析

利用简并PCR和TAIL PCR从嗜热特异腐质霉Humicola insolens Y1中克隆得到一个新的耐热型木聚糖酶基因xynD。该基因全长1 104 bp, 共编码367个氨基酸,不含有内含子,N端的22个氨基酸为信号肽。成熟蛋白在毕赤酵母中进行了重组表达,重组蛋白经超滤和离子交换柱纯化后达到电泳纯。对其酶学性质分析表明,XynD的最适pH为6.0,在pH 4.0～7.0可以保持80%以上的酶活性,在pH 5.0～10.0具有良好的pH稳定性。最适作用温度为70℃,在60℃条件下保持稳定。多数金属离子对XynD酶活力没有明显的影响。麦芽汁降解实验表明,XynD与商用β 葡聚糖共同作用可以提高麦芽的过滤速率(45.7％)并降低粘度(81％)。因此XynD在酿酒业,特别是啤酒制造业具有潜在的应用价值。     

木聚糖酶对不同木聚糖含量的仔猪饲粮养分消化率的影响

试验设计3个消化试验分别研究在3个饲粮木聚糖水平下添加不同剂量的木聚糖酶对养分表观消化率的影响。饲粮木聚糖水平分别为5.33%、6.47%和8.44%。每个消化试验(即每个木聚糖水平下)设5个处理,分别饲喂不添加酶的基础饲粮和添加4个木聚糖酶水平的饲粮。随着饲粮木聚糖含量的提高,木聚糖酶的添加量相应增加。结果表明:当饲粮木聚糖含量低于6.47%时,添加木聚糖酶对养分消化率没有改进作用,而当木聚糖含量高达8.44%时,木聚糖酶的添加对养分消化率有一定的改进效果。与不加酶组比,加酶组粗蛋白、有机物、钙、磷、酸性洗涤纤维和能量的平均消化率分别提高了2.92%、1.05%、5.80%、5.15%、2.36%和0.84%。     

Effects of beta-glucanase and xylanase supplementation on gastrointestinal digestive enzyme activities of weaned piglets fed a barley-based diet

The effects of supplementing a barley-based diet for weaned piglets withexogenous beta-glucanase and xylanase on gastrointestinal digestiveenzyme activities were investigated. Thirty-six cross-bred weaned pigletswere randomly assigned to two groups with three pens based on sexand mass. Each group was fed on the diet based on barley with or withoutadded beta-glucanase and xylanase (0.15%) for a 4-week period. Theresults showed that enzyme supplementation improved growth performanceof piglets significantly (p < 0.05), but had no effect (p = 0.091)on average daily feed intake. The results also showed that supplementationof beta-glucanase and xylanase had no effect on pepsin activity in gastriccontents but slightly decreased (p = 0.092) the pepsin activity ingastric mucosa. Meanwhile, no effect of enzyme supplementation ontrypsin activity in duodenal contents was observed. However, the activitiesof amylase and lipase in duodenal contents were significantly(p < 0.05) decreased, whereas the activities of maltase, sucrase andgamma-glutamyl transpeptidase (gamma-GT) in jejunal and ileal mucosa wereenhanced significantly (p < 0.05). The improvement of disaccharidaseand gamma-GT activity may be attributed to the positive impacts of exogenousenzymes on digestion and absorption of the nutrients. In conclusion,the current results indicated that supplementation with enzymes in barley-based diets could improve the growth performance of piglets,decrease the activities of amylase and lipase in duodenal contents andincrease the activities of disaccharidase and gamma-GT in jejunal and ilealmucosa.     

绿汁发酵液、纤维素酶和木聚糖酶对灵芝菌糟发酵饲料品质的影响

本试验旨在研究绿汁发酵液、纤维素酶和木聚糖酶对灵芝菌糟发酵饲料品质的影响,为利用菌糟生产优质发酵饲料提供理论依据.试验分为7个处理组,以添加蒸馏水为对照(CON组),灵芝菌糟中分别添加2 mL/kg绿汁发酵液(FGJ组)、2000 U/kg纤维素酶(CEL组)、5000 U/kg木聚糖酶(XYL组)、2 mL/kg绿汁发酵液+2000 U/kg纤维素酶(MIX1组)、2 mL/kg绿汁发酵液+5000 U/kg木聚糖酶(MIX2组)、2 mL/kg绿汁发酵液+2000 U/kg纤维素酶+5000 U/kg木聚糖酶(MIX3组),每个处理组3个重复,常温下发酵60 d开封,测定灵芝菌糟发酵饲料的化学成分和发酵品质.结果表明,与CON组相比,FGJ组、CEL组和XYL组可溶性碳水化合物(WSC)含量极显著升高(P<0.01),pH、气体损失率(GLR)及中性洗涤纤维(NDF)、半纤维素(HC)、氨态氮(AN)含量均极显著降低(P<0.01);与FGJ组相比,MIX1组和MIX2组乳酸(LA)、乙酸(AA)含量显著或极显著升高(P<0.05;P<0.01),WSC含量极显著升高(P<0.01),AN含量极显著降低(P<0.01).与CEL组相比,MIX1组的干物质回收率(DMR)及NDF含量极显著降低(P<0.01),WSC、AN含量极显著升高(P<0.01).与MIX1组和MIX2组相比,MIX3组pH、GLR极显著降低(P<0.01),酸性洗涤纤维(ADF)含量显著降低(P<0.05).综上所述,添加纤维素酶和木聚糖酶的效果均优于添加绿汁发酵液,但复合添加效果更佳,其中以MIX3组的添加效果最佳.     

体外酶解法评价饲用木聚糖酶的应用研究

在体外环境下,应用酶解法评价了来源不同的饲用木聚糖酶对不同饲料原料的作用效果。试验结果表明,4种酶中酶Ⅱ的作用效果最好。结果表明,体外酶解试验可以有效的评价饲用酶制剂的作用效果,为针对不同饲料配方合理选择饲用酶制剂提供指导。     

Assessment of Dietary Rovabio Excel in Practical United States Broiler Diets

The focus of this study was to evaluate a feed enzyme (Rovabio Excel) in diets fed to commercial broilers reared in environments typical to that observed in the US broiler industry and destined to be marketed at BW at or near 2.5 kg. The feed enzyme was tested in diets differing in amino acid and energy composition. The main effect of altered nutrient levels, mainly reduced amino acids, was higher abdominal fat at processing. The formulation addition of the exogenous feed enzyme to diets differing in nutrient contents and energy did not affect broiler live performance or carcass traits. However, the exogenous feed enzyme decreased 14-d mortality in experiment 1 and 41-d mortality in experiment 2 but was without effect on mortality in experiment 3. Future research elucidating any benefits on mortality from the exogenous feed enzyme may aid commercial poultry producers. Further, reducing dietary nutrient levels beyond that considered in this study may allow for exogenous enzyme benefits in broiler live production and carcass traits to be observed as mediated through nutrient liberation.     

仔猪饲粮木聚糖水平与添加木聚糖酶对养分消化率的影响

本研究包括2个消化试验,分别研究仔猪对饲粮木聚糖的耐受能力和添加木聚糖酶的效果。试验1:设计5个饲粮木聚糖水平。试验2:以含7.25%木聚糖饲粮为基础,分别添加0、720、1 440、1 880、5 760 U/kg木聚糖酶。结果表明:随着饲粮木聚糖含量的增加,能量、粗蛋白、酸性洗涤纤维、有机物、钙和磷的消化率下降;当木聚糖含量为8.44%时,养分消化率的降低达到了显著或极显著水平。回归分析表明,养分消化率与饲粮木聚糖含量之间呈较强的负相关,木聚糖酶的添加可以提高饲粮的养分消化率。由此得出:约15 kg体重的仔猪对饲粮木聚糖具有较强的耐受力,随着木聚糖含量的增加,饲粮养分的消化率呈不同幅度的下降;当木聚糖含量达到或高于7.25%时,添加木聚糖酶对养分消化率有一定的改进效果。     

玉米-杂粕型日粮添加木聚糖酶对肉鸡免疫功能、肠道形态和微生物菌群的影响

试验研究在玉米-杂粕型日粮中添加木聚糖酶对肉鸡免疫功能、肠道形态及微生物菌群的影响。试验将480只1日龄艾维因肉公鸡分为4个处理组,分别在基础日粮中添加0、709、1 419、2 128 IU/kg的木聚糖酶,每个处理6个重复,每个重复20只肉鸡,试验期44 d。结果表明:在玉米-杂粕型日粮中添加木聚糖酶对肉鸡免疫器官指数和免疫功能有一定影响1,419 IU/kg组44日龄肉鸡血液E花环和淋巴细胞转化率显著高于对照组(P<0.05);添加木聚糖酶可不同程度提高22日龄肉鸡空肠和回肠肠道肠绒毛长度,降低十二指肠和回肠隐窝深度,显著增加肉鸡肠绒毛长度和隐窝深度的比值(P<0.05);显著降低十二指肠固有层厚度(P<0.05);显著增加44日龄肉鸡乳酸杆菌的数量。结果显示,在玉米-杂粕型日粮中添加1 419 IU/kg水平的木聚糖酶对改善肉鸡免疫指标、肠道形态及微生物菌群效果最佳。