产纤维素酶菌群的筛选及其酶学特性研究

从常年堆放的腐质豆秆中分离到1组产纤维素酶菌群MO,并在50℃、pH8.0条件下培养,90h时达到最高酶活,活性为1.3611IU。该酶最适反应温度为60℃,pH为7.0,在60℃以下和pH3~8范围内具有良好的热稳定性和pH稳定性。在最适反应条件下,该酶的最高活性可达2.13IU。通过生长动力学研究了菌群内部不同生长阶段pH、酶活和失重率的相互关系,并通过非变性电泳对酶谱进行初步研究,得到7条活性条带,说明MO中具有多种产酶菌株。     

健康家鱼肠道细菌中的嗜水气单胞菌及其胞外酶分布

从健康鲢、鲤、草鱼和团头鲂肠道食物和肠壁样品中分离出１８５株细菌菌株，利用糊精一品红培养基、氧化酶测定法和淀粉酶测定法对其中的嗜水气单胞菌颁作了调查，鉴别出１９株嗜水气单胞菌、对这批菌株进一步进行了蛋白酶、ＣＭＣ酶、滤纸酶和溶血毒素调查，发现肠道嗜水气单胞菌分别产蛋白酶和ＣＭＣ酶，但均不产滤纸酶和溶血毒素。本文结果证明了肠道嗜水气单胞是肠道正常微生物鲜落之一，对鱼体有一定的有利作用。     

皱马鞍菌液体培养过程中胞外酶及还原糖的动态变化

对皱马鞍菌(Helvella crispa)液体培养过程中不同胞外酶活性和还原糖含量的动态变化进行了研 究。结果表明,在28 ℃、110 r,／min恒温振荡培养条件下,皱马鞍菌菌丝体生物量在第13天达峰值(9．40 g／L);发酵 液pH值逐渐上升,从第9天(8．24)后基本保持平稳;胞外蛋白含量于第6天达峰值(8．30 mg／mL);胞外还原糖 含量急剧上升,至第3天达峰值(20．43 mg／mL),之后迅速下降,到第8天降为1．38 mg／mL,此后一直维持在1 mg／mL水平。皱马鞍菌对碳水化合物的利用顺序为淀粉、纤维素、木质素。淀粉酶的活性高峰出现在第3天,并一 直保持在较高水平;CMC酶6 d后急剧分泌,第7天达峰值,之后又急速下降;愈创木酚氧化酶和多酚氧化酶活性 高峰出现最晚,均在第14天。酸性蔗糖转化酶活性在培养期间始终很低,而中性蔗糖转化酶活性呈阶梯上升趋势。     

碱性纤维素酶产生菌的筛选及产酶条件的研究

[目的]筛选碱性纤维素酶产生菌,并优化其产酶条件。[方法]对从土壤样品中分离到的100余株菌进行平板筛选,获得1株产碱性纤维素酶的菌株ZJJ-1,并对其进行液体培养基成分及发酵产酶条件优化。[结果]培养基最佳配方为：麸皮0.5%,大豆粉2%,KH2PO40.2%,NaCl 0.7%;最优产酶条件为：37℃、175 r/min培养48 h,初始pH值为7。在此条件下,最高酶活水平达51.20 U/ml。[结论]为碱性纤维素酶的后续研究提供了优良的菌种资源。     

里氏木霉产纤维素酶条件的优化

研究采用里氏木霉菌株30911、40358和40359,设计了9个影响里氏木霉产纤维素酶活性因素,对里氏木霉的纤维素酶活性进行了液体摇瓶发酵试验。结果表明,培养基中微晶纤维素和小麦麸皮的最适添加量分别为:微晶纤维素20 g·L-1,小麦麸皮80 g.L-1,微晶纤维素与小麦麸皮最适配比为1:4;接种孢子悬液浓度1×107个·mL-1,培养温度28～30℃,pH 5.5,培养时间72 h,摇瓶转速180 r·min-1,250 mL三角瓶中装液量为50～75 mL。     

自然条件下纤维素分解真菌的分离筛选

以自然界中腐烂的枝条、朽木为材料,经过多代分离、筛选,获得4株可降解利用纤维素的真菌,分别标记为F-1、F-2、F-3和F-4.生长速率和纤维素酶活性检测结果表明,4株真菌都能较好地利用培养基中的纤维素类物质,在以羧甲基纤维素钠（CMC—Na）或滤纸为唯一碳源的平板培养基上长势良好,能使滤纸快速裂解;4株真菌对蔗糖的利用效果好于葡萄糖,适宜的富集增殖培养基为马铃薯蔗糖琼脂培养基（PSA）。在MS无机盐添加羧甲基纤维素钠的发酵培养基中,各菌株具有不同的产纤维素酶能力,其中F-1的羧甲基纤维素（CMCase）酶活最高,F-4的滤纸酶活力（FPase）最高;在单独的杂草、木屑培养基上,4株真菌均能迅速生长并大量增殖,其中以F-1和F-3生长速率最快。因此,初步认为4株真菌分解纤维素类物质的能力都较强。     

In vitro Evaluation of Feed Enzyme Compound Produced by Aspergillus sulphureus in Solid-state Fermentation

Three trials were conducted to analyze a multi-enzyme compound produced by Aspergillus sulphureus in solid-state fermentation (SSF) as a potential feed additive.The results of the first trial showed that there were at least 5 non-starch polysaccharide enzymes:xylanase,β-glucanase,pectinase,mannase and carboxy methyl cellulase (CMCase) contained in the compound.Xylanase and β-glucanase showed good activities at pH 2.5-7.0,which were in the range of 649-1046 U/g and 444-648 U/g,respectively.Pectinase showed good activity in acidic solution (pH 2.5-3.0),which ranged from 195 to 917 U/g.Mannase showed high activity of 235-298 U/g at pH 3.5-4.5 and the activity of CMCase was relatively constant at pH 2.5-7.0,which was in the range of 38.2-78.6 U/g.The second trial was aimed to test the stability of the enzymes in gastric liquor (pH 2.6) of finishing pigs and Na2HPO4-gastric liquor (pH 5.5).After 6 h incubation at 40℃ in gastric liquor,the retained activity of xylanase,β-glucanase,pectinase,mannase and CMCase was 26.3%,65.0%,71.0%,74.8% and 85.6%,respectively.While after 6 h incubation at 40℃ in Na2HPO4-gastric liquor,the retained activity of xylanase,β-glucanase,pectinase,mannase and CMCase was 87.9%,91.1%,92.3%,95.0%,and 97.5%,respectively.The third trial was carried out in a jejunum liquor (pH 5.8,200 mL),which contained 0.2 g of the multi-enzyme compound and 10 g of soybean hull or wheat bran,respectively.After 8 h incubation at 40℃,18.7% of soybean hull and 20.1% of wheat bran could be degraded to soluble saccharide,respectively.Compared with the traditional methods for feed enzyme testing which involve feeding animals for 1-3 months,enzyme assay in this way was relatively convenient.     

姬松茸菌株的胞外酶活性及与产量的关系

研究了姬松茸(Agaricus blazei Murrill)菌株(出发菌株AbM9,AbM9经离子束注入选育的菌株AbML2、AbML7和AbML11,及AbM9单孢分离所得菌株AbMD3)在菌丝生长期、菇蕾期、一潮菇期和二潮菇期培养料的胞外羧甲基纤维素酶、淀粉酶和蛋白酶活性的动态变化及与产量的关系。试验结果表明,姬松茸菌株AbML11产量最高,与AbML2、AbMD3、AbML7和AbM9的差异达极显著水平,但AbML11的产量与菌丝长速相关性不显著;5个供试菌株的淀粉酶活性在菌丝生长期均最高,与菌丝长速的相关性不显著;蛋白酶活性在菇蕾期最高,与产量的相关性不显著;羧甲基纤维素酶活性在一潮菇期最高,与产量呈正相关,相关系数为0.928,达显著水平。本研究可为姬松茸高产菌株的筛选提供一定的理论依据。     

绵羊瘤胃微生物CMCase酶促反应最适条件研究

试验系统地测定了绵羊瘤胃微生物细胞内和细胞外羧甲基纤维素酶(CMCase)酶促反应的最佳条件,包括最适pH值、底物用量、酶量、反应时间和反应温度。结果发现:当pH值介于4.5～6.0之间时,CMCase活力无显著差异,其中胞外CMCase在pH值为5.5时活力最高,胞内CMCase在pH值为5.0时活力最高;随着底物用量的增加,酶活力极显著增加(P<0.01),并在10mg时达到最大值;酶量在0.25ml时,胞内CMCase活力达到最大值,而胞外CMCase则在0.5ml时最高;随着反应时间的延长,CMCase活力均逐渐下降,无论是胞外还是胞内CMCase,反应10min时即达到最高活力,且在胞外同其它组比较差异极显著(P<0.01),在胞内显著高于反应30～60min时的活力;胞内和胞外CMCase活力均在45℃时最高。     

Utilization of pretreated bagasse for the sustainable bioproduction of cellulase by Aspergillus nidulans MTCC344 using response surface methodology

Response surface methodology (RSM) was used to optimize the conditions for the production of endo β-1,4 glucanase, a component of cellulase by Aspergillus nidulans MTCC344 under solid state fermentation, using bagasse as the chief substrate. A four-factor-five-level central composite design was employed for experimental design and analysis of the results. Maximum cellulase activity (CMCase was 28.96 U g⁻¹) can be attained at the optimum conditions, 16.8 mm bagasse bed height, 60% moisture content, pH 4.25 and temperature 40 °C in the solid state fermenter. These data were rather close to the experimental results obtained (CMCase was 28.84 U g⁻¹). A. nidulans MTCC344 was able to hydrolyze pretreated bagasse completely after 8 days of incubation with significant endo β-1,4 glucanase activities. The results of Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) of bagasse showed structural changes through pretreatment, in favor of enzymatic hydrolysis. Bagasse with alkali pretreatment using sodium hydroxide is a source of lignocelluloses able to improve the yield of endo β-1,4 glucanase by the strain of A. nidulans. The endo β-1,4 glucanase produced during the bioconversion of cellulose to glucose by A. nidulans MTCC344 is strongly dependent on the pretreatment given before hydrolysis.