纤维素酶产生菌灰白青霉HML0233的分离及系统分类研究

通过滤纸纤维素平板及发酵产酶鉴定,从腐木下土壤中分离出一株分解纤维素能力较强的真茵HM10233.该菌株的CMC酶活为31.04 U/mL,β-葡萄糖苷酶酶活为47.66 U/mL,滤纸酶活为5.28 U/mL.根据形态特征观察初步推测该菌株属于青霉属;经5.8S rDNA基因两侧的内转录间隙ITS序列同源性分析及构建系统进化树分析.该菌株和Penicillium canescens NKRL 35656同源性很高,推测该菌株为灰白青霉(Penicilliumcanescens).     

产纤维素酶菌株的筛选及酶学性质的研究

从牛粪有机肥中筛选获得1株产纤维素酶的菌种,刚果红法鉴定该菌株具有一定的水解纤维素的能力.显微镜镜检及菌落表型观察,初步鉴定该菌株为芽孢杆菌属细菌;对菌株所产纤维素酶进行酶学性质的研究发现该酶的酶促反应最佳作用温度是45℃,最佳作用pH为4.5～5.5.     

高产脂肪酶菌株的分离鉴定及其酶学性质研究

本研究旨在筛选出一株高产脂肪酶的菌株并进行鉴定,同时评定所产脂肪酶的酶学性质,为饲用脂肪酶的开发提供理论依据和实用参考。通过平板法筛选产酶菌株,根据菌株形态、培养特征、生理生化试验和ITS-r DNA序列分析确定其属种,并对产酶菌株进行初步的酶学性质研究。从富含油污的土壤中筛选出一株高产脂肪酶菌株FZ-4,并鉴定为白地霉(Galactomyces geotrichum),其初步酶学性质研究表明,该菌株所产脂肪酶为胞外酶,在28℃、250 r/min条件下培养96 h所产脂肪酶活力达16.7 U/m L,该酶最适反应p H为7.5,最适反应温度为40℃,在p H为7.0~9.0和20~45℃具有较好的稳定性,且该酶对C16脂肪酸有较强的底物专一性。分离获得的Galactomyces geotrichum FZ-4是一株高效的产脂肪酶菌株,具有潜在的研究和开发价值。     

纤维素酶高产菌株的选育

从土样中分离筛选出12株产纤维素酶能力较强的菌株。经摇瓶发酵复筛,获得产纤维素酶活较高且稳定的菌株F6,其CMC酶活为887 U/mL,滤纸酶活为210.72 U/mL。以该菌株为出发菌株,经紫外线诱变处理,获得纤维素酶高产菌株4-2-2。将菌株4-2-2摇瓶发酵4 d后,产CMC酶活达3171.598 U/mL,滤纸酶活达1827.372 U/mL,分别比出发菌株提高3.58倍和8.67倍。     

玉米秸秆高效分解菌株的筛选、鉴定及产酶条件优化

为了从瘤胃液中筛选出一株对玉米秸秆分解能力较强的纤维菌,试验采用稀释平板法分离奶牛瘤胃菌,通过刚果红平板法对产纤维素酶的菌株进行初筛,选取透明圈大的菌株进行玉米秸秆摇瓶复筛,得到一株对玉米秸秆分解率最高的菌株。根据菌株形态、生理生化特征对菌株进行初步鉴定,并对该菌株的产酶条件进行了优化。结果表明,该菌株为芽孢杆菌属,接种量为8%、氮源为NH3NO3、吐温80含量1%为该菌最适产酶条件。     

鹅肠道纤维素分解菌的分离鉴定及其产酶条件的优化

本研究拟在鹅肠道筛选1株高效降解纤维素的菌株,并对该菌株的产酶条件进行研究.通过对鹅肠道菌群进行富集培养、分离纯化,利用刚果红法(初筛)和摇瓶法(复筛)得到1株产酶活较高的纤维素分解菌E2.经16S rDNA核苷酸序列比对分析表明,该菌株为芽孢杆菌属的短小芽孢杆菌.单因素试验得出菌株E2的产酶最适碳源为玉米粉,最适氮源...     

B .methylotrophicus SWU6菌株产纤维素酶发酵条件的优化?

前期从自然界土壤中分离筛选到一株纤维素酶产生菌 Bacillus methylotrophicus SWU6菌株,为提高该菌株发酵产酶能力,本研究采用3,5二硝基水杨酸显色法(DNS)对该菌株产酶所需碳源、氮源、温度、pH 值、装瓶量、接种量等发酵条件进行优化,并比较优化前后等量发酵上清液中 CMCase 酶活大小。结果表明：SWU6菌株产纤维素酶的最适碳源为淀粉,最适氮源为牛肉膏,最适培养温度为50℃,培养基最适初始 pH 值为5.0,最适装瓶量为20%,最适接种量为2%;在最优发酵产酶条件下,SWU6菌株发酵上清液中的 CMCase 酶活达到454.69 U/mL,明显高于优化前利用基础培养基发酵所产的 CMCase 酶活,且酶活提高约3倍。通过发酵条件优化后,SWU6菌株单位体积发酵液显示出了更强的纤维素酶活性。     

高产淀粉酶枯草芽胞杆菌的诱变选育和酶学性质研究

为了提高产淀粉酶枯草芽胞杆菌Bacillus subtilis N21的酶活,试验采用紫外线对该菌株进行诱变,并对该淀粉酶的部分酶学性质进行了研究。结果表明,淀粉酶酶活由出发株的32.96 U/mL增加到86.24 U/mL,比原菌株酶活提高了161.65%。该酶的最适反应温度为35℃,最适pH值为7.0,在30~40℃,pH值为5.0~8.0条件下较稳定。Ca2+和Mn2+对酶有激活作用,Cu2+、Zn2+对酶有抑制作用,而Mg2+、Fe2+对其影响较小。说明筛选出1株产酶活性较强的突变菌株。     

饲用酸性蛋白酶产生菌肉桂色曲霉9824菌株的筛选研究

试验研究了饲用酸性蛋白酶产生菌分离筛选的方法,筛选得到肉桂色曲霉R-24、黑曲霉H-17和宇佐美曲霉Y-19三株酶活力较高的菌株。三株菌所产酶的最适作用温度为40℃和pH值为3。经综合比较,选择肉桂色曲霉R-24为产酶菌株,将其诱变得到肉桂色曲霉9824变异株,并研究了该菌株的产酶条件和酶热稳定性。     

青藏高原低温纤维素降解菌的筛选与酶学特性

以青藏高原冷地早熟禾草地土壤为样品,筛选低温纤维素降解菌株。采用刚果红染色法(水解圈法)和3,5-二硝基水杨酸(DNS)法进行菌株筛选与酶学特性研究,结合形态学与分子生物学方法对其进行鉴定。筛选出一株低温纤维素降解菌株(LD19),经鉴定为钉斑链霉菌(Streptomyces clavifer),同时该菌还具有降解半纤维素的能力。该菌产纤维素酶的最适温度为40℃,最适pH值为6,在30～70℃、pH值4～10时相对酶活力较稳定,Zn~(2+)对酶有抑制作用。该菌产半纤维素酶的最适温度为50℃,最适pH值为6,在30～40℃、pH值4～9时相对酶活力较稳定,Cu~(2+)等对该酶有促进作用。该菌株在培养102 h后两种酶的相对酶活力均达到最高值。菌株LD19可以同时分泌低温纤维素酶和半纤维素酶,在饲用酶制剂的应用中具有较大潜力。     

不同来源木聚糖酶对黄羽肉鸡屠宰性能和免疫功能的影响

为研究小麦型日粮中添加不同来源木聚糖酶对黄羽肉鸡屠宰性能和免疫功能的影响,选择22日龄体重相近的健康黄羽肉鸡375羽,随机分成3个处理,每处理5个重复,每重复25羽。对照组饲喂小麦型基础日粮,试验Ⅰ、Ⅱ组分别饲喂在基础日粮中添加250 mL/t真菌性和细菌性木聚糖酶的试验日粮。试验期为28 d。结果表明,小麦型日粮中分别添加250mL/t真菌性和细菌性木聚糖酶显著提高了黄羽肉鸡腹脂率（P〈0.01）,而对全净膛率、胸肌率和腿肌率没有影响（P〉0.05）;显著提高了黄羽肉鸡法氏囊指数（P〈0.01）以及血清中T3（P〉0.05）、T4、IgA（P〈0.01）和IgM（P〈0.01）水平,且均表现出试验Ⅱ组〉试验Ⅰ组〉对照组的规律,但脾脏指数却相反。结果提示,两种来源木聚糖酶可提高黄羽肉鸡腹脂率和增强机体免疫功能,且细菌性木聚糖酶的提高幅度均高于真菌性木聚糖酶。     

木聚糖酶对小麦中非淀粉多糖体外酶解效果的影响研究

为了探究木聚糖酶对小麦非淀粉多糖的降解效果。试验采用单因子设计,用气相色谱法测定不同木聚糖酶水平,在一定水分(料水比为1:10)、温度(40℃)和pH值(6.0)条件下酶解2 h,小麦非淀粉多糖含量变化。在酶水平为每克小麦60 000 IU时,降解效果最大;以可溶性非淀粉多糖、不可溶性非淀粉多糖、总非淀粉多糖含量变化为指标,每千克小麦的适宜木聚糖酶添加量分别为0.718 g、0.732 g、0.717 g。添加木聚糖酶对小麦非淀粉多糖有显著的降解效果。     

不同来源木聚糖酶及其组合对木聚糖水解产物组成、细菌增殖与大肠杆菌黏附性的影响

本试验旨在研究不同来源木聚糖酶及其组合对木聚糖的水解效果,并研究不同木聚糖水解产物对细菌增殖及大肠杆菌对肠道上皮细胞黏附性的影响。试验用木聚糖酶A和木聚糖酶B分别来源于毕赤酵母和米曲霉。采用木聚糖酶A、木聚糖酶B、组合酶1(木聚糖酶A∶木聚糖酶B=3∶7)、组合酶2(木聚糖酶A∶木聚糖酶B=1∶1)、组合酶3(木聚糖酶A∶木聚糖酶B=7∶3)分别水解木聚糖,然后测定木聚糖水解产物对大肠杆菌、枯草芽孢杆菌和乳酸杆菌增殖和大肠杆菌对肠道上皮细胞黏附性的影响。结果表明:1)2种木聚糖酶有一定的组合效应,木聚糖酶A、木聚糖酶B、组合酶1、组合酶2、组合酶3组木二糖和木三糖的总含量分别为95.70%、86.79%、93.11%、94.55%和87.55%,其中木聚糖酶A组木二糖含量最高,组合酶2组木三糖含量最高。2)培养20 h时,5种木聚糖水解产物对大肠杆菌的增殖(以菌液吸光度值表示)没有产生显著影响(P0.05);培养20和30 h时,5种木聚糖水解产物显著促进枯草芽孢杆菌的增殖(P0.05);培养13和17 h时,5种木聚糖水解产物显著促进乳酸杆菌的增殖(P0.05)。3)5种木聚糖水解产物均可以显著降低大肠杆菌对肠道上皮细胞的黏附率(P0.05)。由此可见,通过不同来源木聚糖酶及其组合水解木聚糖,可以产生以木二糖和木三糖为主要组分的水解产物,从而起到促进枯草芽孢杆菌和乳酸杆菌增殖、减少大肠杆菌对肠道上皮细胞黏附的作用。     

变温操作对黑曲霉A-25产木聚糖酶的影响

试验研究不同温度对黑曲霉生物量和木聚糖酶产量的影响,并通过单位质量菌丝的产酶量和酶谱探究木聚糖酶产量提高的原因。结果显示:黑曲霉A-25在33℃培养时获得最大的生物量,而在29℃时获得最高胞外木聚糖酶分泌量。因此,变温操作采用0~48 h在33℃下培养、之后降为29℃,木聚糖酶活力可达721 U/mL,比29℃恒温发酵提高18%,同时发酵周期缩短8 h。进一步研究发现,0~48 h阶段内变温发酵条件下黑曲霉菌丝体生物量明显增加,并且单位质量菌丝体的产酶能力也有不同程度的提高;木聚糖酶的酶谱检测发现,变温发酵促进木聚糖酶XynⅢ的提前表达,同时提高木聚糖酶XynⅠ和木聚糖酶XynⅡ的表达量。     

1株产碱性木聚糖酶的芽胞杆菌的分离鉴定及其相关研究

分离到1株产碱性木聚糖酶的革兰氏阳性菌株,通过菌落形态及16srDNA序列同源性分析,确定该菌株为短小芽孢杆菌。生长条件和产酶条件研究结果显示,最适温度和pH分别为50℃和pH8·0。培养基优化试验显示在有机和无机混合氮源条件下(NH4NO30·57%;牛肉膏1%;蛋白胨0·5%;酵母提取物0·5%;木聚糖0·5%)木聚糖酶产量达到最高(180U/mL)。酶学试验表明最适反应条件为50℃,pH8~9;在pH9的条件下,孵育120min时仍具有75%的活力,表明该酶具有较强的碱耐受性。     

木聚糖酶对肉鸡能量饲料养分利用率和表观代谢能值的影响

本试验通过肉鸡代谢试验考察木聚糖酶对玉米、小麦、麦麸和米糠的养分利用率和表观代谢能(AME)值的影响,确定木聚糖酶与不同能量饲料的定量关系.试验包括4个单因子试验,分别用玉米、小麦、麦麸、米糠进行代谢试验,每种饲料设5个处理,添加不同水平木聚糖酶,每个处理8个重复,每个重复1只鸡.试验结果表明:对于玉米,当木聚糖酶添加水平为1 500 IU/kg时,与对照组相比,干物质利用率提高了0.93%(P<0.05).对于小麦,木聚糖酶水平为3 000 IU/kg时,干物质、能量和蛋白质利用率提高幅度最大,分别提高了7.02%(P<0.01)、6.78%(P<0.01)和22.31%(P<0.05).对于麦麸,在木聚糖酶为6 000 IU/kg时,干物质和能量利用率提高幅度最大,分别提高10.14%和14.73%(P<0.01);在木聚糖酶为4 000 IU/kg时,蛋白质利用率提高157.31%(P<0.01).对于米糠,在酶水平为1 500 IU/kg时,米糠的蛋白质利用率提高了27.92%(P<0.05).适宜水平的木聚糖酶显著提高了小麦、麦麸的AME值.结果表明,玉米、小麦、麦麸、米糠中每克总木聚糖的适宜木聚糖酶添加量分别为35.38、27.01、30.21和18.27 IU,每克水溶性木聚糖的适宜木聚糖酶添加量分别为1 250、273.56、753.33和937.5 IU.     

Effect of dietary xylanase on energy, amino acid and mineral metabolism, and egg production and quality in laying hens

1. The aim was to examine the effect of dietary xylanase on the availability of nutrients for laying hens when fed on wheat-rye-soy-based diets. The basal diet was formulated to contain 11.03 MJ/kg apparent metabolisable energy (AME), and the experimental diets were formulated by supplementing the basal diet with four different activities of xylanase (400, 800, 1200 and 1600 xylanase units (XU)/kg). 2. The AME and nitrogen metabolisability coefficients of xylanase-supplemented diets were 1.2% and 7.1%, respectively, greater than in the control diet. 3. Supplementary xylanase significantly improved the coefficients of metabolisability of indispensable, dispensable and total amino acids by 8.2%, 6.9% and 7.8%, respectively, and led to a significantly linear response of total amino acid metabolisability coefficient to xylanase. There was a range of effects within the indispensable amino acids with xylanase supplementation (1600 XU/kg) significantly improving the metabolisability of threonine by 4.9%, but having no have effect on lysine. The response of total amino acid retention to added xylanase was a significant quadratic function and suggests that 800 XU/kg is the optimum supplementary dosage. 4. Supplementary xylanase significantly improved sulphur metabolisability in a linear fashion to a maximum of 2.3% higher than that of the control diet. In terms of daily retention, most of the minerals responded in a quadratic manner to dietary xylanase, as the suggested optimal supplementary level was between 800 and 1200 XU/kg. 5. The yolk colour of the birds receiving 1200 and 1600 XU/kg was 0.33 and 0.28 units (Roche score); these were 4.1% and 3.5%, respectively, darker than the yolk of the birds given the control diet. 6. Birds receiving xylanase had a significantly higher weight gain than those fed on the unsupplemented diet. Feed intake, the number of eggs per hen per d, dirty and cracked eggs, and feed conversion ratio for egg production were not affected by xylanase. These data suggest that use of a xylanase may improve the metabolisability of many nutrients, but that such effects may not always benefit production parameters.     

Influence of phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digestive tract measurements and gut morphology in broilers fed wheat-based diets containing adequate level of phosphorus

1. The aim of the present study was to examine the influence of microbial phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digesta viscosity, digestive tract measurements and gut morphology in broilers fed on wheat-soy diets containing adequate phosphorus (P). The wheat-soy basal diet was formulated to contain 4.5 g/kg non-phytate P and the experimental diets were formulated by supplementing the basal diet with xylanase (1000 xylanase units/kg diet), phytase (500 phytase units/kg diet) or a combination of phytase and xylanase. 2. Supplemental phytase improved the weight gains and feed efficiency by 17.5 and 2.9%, respectively. Corresponding improvements due to the addition of xylanase were 16.5 and 4.9%, respectively. The combination of phytase and xylanase caused no further improvements in broiler performance. 3. Individual additions of xylanase or phytase resulted in numerical improvements in apparent metabolisable energy (AME), but the differences were not significant. The combination of the two enzymes significantly increased AME. Addition of xylanase and the combination of the two enzymes reduced the viscosity of digesta in all sections of the intestine. Phytase supplementation reduced digesta viscosity in the duodenum and ileum, but not in the jejunum. 4. Enzyme supplementation lowered the relative weight and length of the small intestine. Additions of xylanase and phytase reduced the relative weight of the small intestine by 15.5 and 11.4%, respectively, while the corresponding reductions in the relative length of the small intestine were 16.5 and 14.1%, respectively. The combination of phytase and xylanase had no further effects on the relative weight and length of the small intestine compared with the xylanase group. 5. The addition of phytase increased villus height in the duodenum and decreased the number of goblet cells in the jejunum compared with those on the unsupplemented basal diet. Xylanase supplementation tended to increase goblet cell numbers in the duodenum and decreased crypt depth in thejejunum. The combination of phytase and xylanase increased villus height in the ileum and crypt depth in thejejunum and ileum. 6. In summary, the present results showed that the addition of a microbial phytase, produced by solid state fermentation and containing significant activities of beta-glucanase and xylanase, was as effective as xylanase in improving the performance of broiler chickens fed on wheat-based diets containing adequate levels of P. Improved performance with enzyme supplementation was generally associated with reduced digesta viscosity, increased AME, and reduced relative weight and length of small intestine.     

饲用木聚糖酶活力的综合评价

试验采用综合分析方法评价了1种饲用木聚糖酶的活力。评价内容包括耐酸性试验、耐胃液试验、热稳定性试验和实际底物降解试验。在pH5.4～6.8条件下,该酶活力较稳定,保持在5000U/g以上,但随着pH的降低,活力迅速下降;在胃液中作用一段时间后该酶的活力损失很快,1.5h后其活力就下降到原有的37.4%;热稳定性试验中,该酶在90℃的环境中作用5min后,酶活损失不超过5%;实际底物降解试验中,反应4h后,无淀粉麦麸的降解率可以达到5%以上。试验结果表明,以上几个指标能够较为充分、准确的评价该木聚糖酶的可应用性,从而为饲料酶活性的系统评价提供了参考,为饲料加工和动物生产提供了更为可靠的数据。     

小麦基础饲粮中添加木聚糖酶对生长猪生长性能、血清生化指标及肠道微生物菌群的影响

本试验旨在研究在小麦基础饲粮中添加木聚糖酶对生长猪生长性能、营养物质表观消化率、血清生化指标及肠道微生物菌群的影响,探讨适宜的木聚糖酶添加水平。选择80头健康、平均体重40 kg的"杜×长×大"生长猪,随机分为5个处理,每个处理4个重复,每个重复4头猪。正对照组饲喂玉米-豆粕型基础饲粮,负对照组饲喂小麦-豆粕型基础饲粮,木聚糖酶组分别饲喂在负对照基础饲粮上添加500、1 000和2 000 U/kg木聚糖酶的试验饲粮。试验期28 d。结果表明:1)小麦基础饲粮中添加木聚糖酶可极显著提高生长猪平均日增重(P<0.01),显著或极显著提高平均日采食量和营养物质表观消化率(P<0.05或P<0.01),其中2 000 U/kg木聚糖酶组效果最好,平均日增重和平均日采食量分别提高了26.0%和16.5%(P<0.01);2)小麦基础饲粮中添加木聚糖酶显著或极显著提高了生长猪血清葡萄糖、三碘甲腺原氨酸、四碘甲腺原氨酸和D-木糖含量(P<0.05或P<0.01),极显著降低血清尿素氮含量(P<0.01);3)饲粮组成和木聚糖酶可显著影响猪肠道微生物菌群,添加木聚糖酶可维持动物肠道微生物菌群的平衡,2 000 U/kg木聚糖酶组肠道菌群结构和玉米-豆粕饲粮饲喂条件下接近。由此可见,添加2 000 U/kg的木聚糖酶可显著改善饲喂小麦基础饲粮生长猪的生长性能和营养物质表观消化率,维持肠道微生物菌群的平衡。在木聚糖含量为11.2%的生长猪小麦基础饲粮中,木聚糖酶的适宜添加水平为2 000 U/kg。