纤维素酶、半纤维素酶降解膨化玉米秸秆工艺优化

利用纤维素酶和半纤维素酶协同降解经过膨化预处理的玉米秸秆,以提高玉米秸秆相比于单酶解的产糖量。在单因素实验的基础上,以还原糖产率为响应值,通过响应面来优化设计实验。实验数据分析得出,膨化玉米秸秆酶解的最佳工艺为:pH 4.8,液固比13∶1,酶解时间60 h,酶浓度6 g/L,温度51℃。对比纤维素酶单独作用于玉米秸秆的降解效果,双酶协同酶解使酶解液的还原糖产率提高到24%,还原糖产率提高了14.3%。协同酶解的研究为木质纤维素原料的降解提供了一种新的方式。     

降解聚丁二酸丁二醇酯菌株的诱变选育及降解性能

诱变选育是基因资源开发的重要手段之一,为了对降解聚丁二酸丁二醇酯(PBS)菌株的降解性能进行改良,选择具有一定降解PBS能力的尖孢镰刀菌(Fusarium oxysporum)DSCF-8为出发菌株,分别研究了其在5种诱变途径下的突变效应及最佳诱变条件,测定了突变菌株D8B14在不同条件下的降解性能。结果表明,复合诱变途径的高效降解菌株获得率大于单因子诱变途径,分别达到了4.55%和1.30%,但突变株的降解特性遗传不稳定;硫酸二乙酯单因子诱变效应大于紫外线诱变和微波诱变,且得到的突变株较复合诱变得到的菌株具有更强的遗传稳定性,最佳诱变条件为5%的浓度处理40 min。硫酸二乙酯诱变得到的突变株D8B14传代稳定且降解性能优异,其对PBS薄膜的降解率较出发菌株提高了34.49%,粗酶液酶活性提高率为23.86%。该突变株在35℃条件下仍表现出较高的降解能力,对不同的初始pH条件亦具有更强的耐受能力,尤其在pH 4.0或pH 9.0的酸性和碱性条件下,降解率分别提高了13.26%和27.04%。     

纤维素降解菌的筛选及其产酶特性

纤维素降解菌在纤维素类物质降解的过程中起着重要作用。通过滤纸条崩解和液态产酶实验,从腐烂的树叶中筛选出1株对纤维素有较强降解能力的菌株SY2,通过形态学特征以及生理生化反应,初步鉴定该菌株为荧光假单胞菌(Pseudomonas fluorescent)。产酶培养基及培养条件优化的实验结果表明:SY2的最佳产酶培养基为麸皮与秸秆粉的质量比1∶4、(NH4)2SO41%、pH8、固液比1∶2;最佳产酶条件为培养温度30℃、接种量7.5%、培养时间72 h;在酶液中添加Fe2+和VB12可使纤维素酶活力分别达到174.89μmol/(g.min)和165.99μmol/(g.min),提高了33.5%和26.3%。SY2对麸皮、滤纸的降解效果较好,降解率分别达到45.0%和33.3%。     

膨化预处理玉米秸秆的还原糖酶解工艺

应用膨化技术对玉米秸秆进行预处理.在单因素实验基础上,以还原糖转化率为影响值设计正交实验,研究温度、pH、液固比、酶浓度及酶解时间五因素对纤维素酶解过程的影响.得出玉米秸秆糖化酶解最佳工艺条件为:温度48℃,pH 4.5,液固比8:1,酶浓度36.0 U/g,酶解时间25 h；在此工艺条件下还原糖转化率达到28.98％.扫描电镜表征对比观察可看出,膨化后玉米秸秆纤维素酶解充分；结合红外光谱对各组分特征基团分析表明,膨化后的玉米秸秆酶解的纤维素基团特征峰变化更为明显.     

PVA降解菌的复配组合及其紫外诱变处理的研究

对已筛选出的4株PVA降解菌进行不同的组合后测定PVA降解率,发现Z5与Z8菌株混合后在1周内对PVA的去除率达到了所有组合中的最高值52.07%,比单菌株提高了约22%。采用紫外照射的方法对混合菌进行诱变并利用正交实验研究其特性,结果表明,诱变后的混合菌在1周内降解了91.46%的PVA,并且碳源浓度对酶活的影响最为显著。当pH为7,碳源、氮源含量分别为1.5 g/L、0.6 g/L,NaC l浓度为1.5 g/L时,诱变菌株处于最佳生长状态。     

木薯生物质酸解制取还原糖并用于软锰矿的还原浸出工艺

对木薯生物质酸解制备还原糖进行了详细研究,考察了硫酸浓度、温度及木薯生物质添加量的影响。结果表明:木薯淀粉、木薯干片和木薯渣经过硫酸处理,可以产生大量的还原糖。在适宜条件下,木薯淀粉的还原糖得率可以达到95%以上,木薯干片的还原糖得率在85%以上,木薯渣的还原糖得率在60%左右。成本分析表明木薯渣制备吨还原糖的原料成本最低。进行了木薯渣湿法还原浸出软锰矿实验,锰浸出率可达到90%以上。     

一株耐盐柴油降解菌的分离鉴定及其降解性能

从某油田附近受石油污染土壤中分离出一株以柴油为惟一碳源的耐盐菌株LS1。通过对菌株的生理生化特性、菌体的形态观察及16S r DNA基因序列分析鉴定菌株LS1为假单胞菌属(pseudomonas)。该菌株可耐受的最高盐度(Na Cl)和柴油浓度分别为6%~8%和12 000 mg/L。菌株生长的适宜p H和温度条件分别为6.0~8.0和28~36℃。在盐度为6%、p H为7.0、温度为32℃、菌种投加量为10%的条件下,初始浓度为3 000 mg/L的柴油经6 d降解后,去除率可达78.3%,加入适量外加碳源葡萄糖和蔗糖,可使降解率分别提高至92%和90%左右。菌株LS1的耐盐机理可能是通过在细胞内积累甜菜碱以调节菌株细胞内外渗透压平衡。投加甜菜碱可提高耐盐菌LS1在高盐环境下对柴油的降解效率。     

巢湖蓝藻酸提取液提高玉米秸秆的酶解效率

资源化利用是避免富营养化水体打捞得到的蓝藻二次污染的有效方法途径。本研究以巢湖蓝藻为例,利用稀硫酸处理得到酸提取液并将其用于玉米秸秆的酶解过程。以主要成分还原糖和蛋白质的提取量为目标,优化了稀硫酸处理条件(处理时间、温度和硫酸浓度)。结果表明,从巢湖蓝藻提取还原糖的效率随着处理温度和硫酸浓度的升高而升高。在处理温度为125℃,处理时间为2 h,稀硫酸浓度为5%时,提取的还原糖最多为(152.8±12.6)mg/g。稀硫酸处理条件对蛋白质的提取效果影响并不显著。在处理温度为105℃,处理时间为1 h,稀硫酸浓度为5%时,最大提取量为(1.55±0.00)mg/g。提取的藻液中和后可以用于玉米秸秆的酶解过程,同对照相比酶解效率能够提升10%左右。结果表明,藻液中的小分子物质和钙离子是主要的作用组分,推测其和木质素表面官能团结合,从而降低了木质素对纤维素酶的非再生吸附。     

书虱伴生菌中纤维素酶产生菌的筛选、鉴定及最佳产酶发酵条件的优化

从以啃食树皮为生的书虱伴生菌中筛选得到5株产纤维素酶的菌株,编号分别为S2、S6、N10、N11和N12。结合菌株的形态学及16S r DNA序列分析等结果将这些菌株分别鉴定为Bacillus methylotrophicus,Streptomyces sp.,Pseudomans fluorescens,Bacillus sp.及Pseudomans sp.。分别使用单因素分析法和响应面分析法对纤维素酶活最高的S2菌株的产酶发酵条件进行了优化。单因素实验结果显示,B.methylotrophicus S2的最适产酶发酵条件为:0.01 g/m L的CMC碳源、10 g/L蛋白胨的氮源、48 h的发酵时间、28℃、初始pH为7.0,此时总纤维素酶活达到204.37 u/g。然后选取发酵时间、温度和初始pH作为3个因素,通过BBD实验,用响应面法对S2的发酵条件进行优化分析,最后得到一个拟合度良好的二次多项方程模型(R2=0.9948)。方差分析结果显示,发酵温度与培养基初始pH之间的交互作用极显著。响应面分析优化后的反应体系为:温度24℃、初始pH为7.7、发酵59 h,测得酶活力为303.18 u/g,比优化前单因素最佳纤维素酶活力提高45%。     

苯胺降解菌的分离和降解特性研究

通过驯化富集培养,从白洋淀底泥中分离筛选出数株能够有效降解苯胺的菌株,经过反复筛选,得到一株能够以苯胺为唯一碳源、高效降解苯胺的菌株BA-1-3.其利用苯胺的最适pH值为7.0,最适温度为30℃,在苯胺浓度为1000 mg/L,180 r/min条件下振荡培养60 h,降解率达到80%以上.经鉴定,菌株BA-1-3属苍白杆菌属(Ochrobactrumsp.).     

二次纤维稀酸水解糖化的研究

研究了稀酸作用下二次纤维的水解糖化,探讨了水解各因素温度、H2SO4浓度、时间、液固比以及不同酸、催化剂对水解效果的影响。实验结果表明,在H2SO4作用下最佳工艺为:H2SO4质量分数4%,水解温度190℃,水解时间40min,液固比15(mL/g);还原糖得率为33.26%,水解率为56.83%。不同稀酸对OCC的水解,HCl比H2 SO4、H3 PO4、HNO3具有明显的优势;催化剂FeSO4与CuSO4对还原糖得率有显著的促进作用,还原糖得率分别提高了39.66%与22.03%,而Al2(SO4)3对OCC水解没有促进作用。最后,对不同水解时间后的OCC残渣的红外结晶指数以及基团结构变化进行了分析,分析结果与实验结果相一致。     

烟草下脚料发酵制取乙醇

通过单因素实验考察了硫酸浓度、固液比和水解时间对硫酸水解的影响。结果显示最优条件为:硫酸浓度为50%(w/w),固液比为10%(w/v),时间为100 min。烟草下脚料在最佳硫酸水解条件下,经5倍稀释,中和pH值至5～6。取经过滤后的水解液(FH)用酿酒酵母(Sacchharomyces cerevisiae)发酵产生乙醇,最大的乙醇浓度和乙醇产量分别为1.09g/L和54.5 g/kg。未过滤水解液(UFH,包括水解残渣)加入纤维素酶(70 U/100 mL)和酿酒酵母(Sacchharomyces cerevisiae)进行发酵,最大的乙醇浓度和乙醇产量分别为1.23 g/L和61.5 g/kg。     

烟草下脚料发酵制取乙醇

通过单因素实验考察了硫酸浓度、固液比和水解时间对硫酸水解的影响。结果显示最优条件为：硫酸浓度为50%（w/w）,固液比为10%（w/v）,时间为100 min。烟草下脚料在最佳硫酸水解条件下,经5倍稀释,中和pH值至5～6。取经过滤后的水解液（FH）用酿酒酵母（Sacchharomyces cerevisiae）发酵产生乙醇,最大的乙醇浓度和乙醇产量分别为1.09g/L和54.5 g/kg。未过滤水解液（UFH,包括水解残渣）加入纤维素酶（70 U/100 mL）和酿酒酵母（Sacchharomyces cerevisiae）进行发酵,最大的乙醇浓度和乙醇产量分别为1.23 g/L和61.5 g/kg。     

Use of fungi for the bioconversion of rice straw into cellulase enzyme

Cellulase production was carried out by solid state bioconversion (SSB) method using rice straw, a lignocellulosic material and agricultural waste, as the substrate of three Trichoderma spp. and Phanerochaete chrysosporium in lab-scale experiments. The results were compared to select the best fungi among them for the production of cellulase. Phanerochaete chrysosporium was found to be the best among these species of fungi, which produced the highest cellulase enzyme of 1.43 IU/mL of filter paper activity (FPase) and 2.40 IU/mL of carboxymethylcellulose activity (CMCase). The "glucosamine" and "reducing sugar" parameters were observed to evaluate the growth and substrate utilization in the experiments. In the case of Phanerochaete Chrysosporium, the highest glucosamine concentration was 1.60 g/L and a high concentration of the release of reducing sugar was measured as 2.58 g/L obtained on the 4th day of fermentation. The pH values were also recorded. The range of the pH was about 5.15 to 5.56 in the case of Phanerochaete Chrysosporium.     

Use of fungi for the bioconversion of rice straw into cellulase enzyme

Cellulase production was carried out by solid state bioconversion (SSB) method using rice straw, a lignocellulosic material and agricultural waste, as the substrate of three Trichoderma spp. and Phanerochaete chrysosporium in lab-scale experiments. The results were compared to select the best fungi among them for the production of cellulase. Phanerochaete chrysosporium was found to be the best among these species of fungi, which produced the highest cellulase enzyme of 1.43 IU/mL of filter paper activity (FPase) and 2.40 IU/mL of carboxymethylcellulose activity (CMCase). The “glucosamine” and “reducing sugar” parameters were observed to evaluate the growth and substrate utilization in the experiments. In the case of Phanerochaete Chrysosporium, the highest glucosamine concentration was 1.60 g/L and a high concentration of the release of reducing sugar was measured as 2.58 g/L obtained on the 4th day of fermentation. The pH values were also recorded. The range of the pH was about 5.15 to 5.56 in the case of Phanerochaete Chrysosporium.     

TiO2光催化氧化降解乳酸的工艺及机理研究

以TiO2为催化剂、紫外灯为光源对乳酸进行光催化降解实验,考察了乳酸初始浓度、TiO2用量、反应时间、曝气方式等因素对乳酸降解率的影响,并在此基础上应用正交实验对降解条件进行优化,同时对乳酸的降解机理进行了探索研究。实验结果表明：以300 W紫外汞灯为光源,在乳酸初始浓度为0.5 g/L、TiO2量为0.20 g/L、反应时间120 min、持续通入空气鼓泡的条件下,乳酸降解效果最佳,乳酸降解率为99.9%;降解12 h总有机碳去除率达91.2%。乳酸光催化降解的反应途径为：乳酸脱羧生成乙醇,乙醇被氧化生成乙醛,进而氧化为乙酸,所有的中间产物被继续降解,最终矿化为CO2和H2O等小分子物质。     

Fumaric acid production using renewable resources from biodiesel and cane sugar production processes

The microbial production of fumaric acid by Rhizopus arrhizus NRRL 2582 has been evaluated using soybean cake from biodiesel production processes and very high polarity (VHP) sugar from sugarcane mills. Soybean cake was converted into a nutrient-rich hydrolysate via a two-stage bioprocess involving crude enzyme production via solid state fermentations (SSF) of either Aspergillus oryzae or R. arrhizus cultivated on soybean cake followed by enzymatic hydrolysis of soybean cake. The soybean cake hydrolysate produced using crude enzymes derived via SSF of R. arrhizus was supplemented with VHP sugar and evaluated using different initial free amino nitrogen (FAN) concentrations (100, 200, and 400 mg/L) in fed-batch cultures for fumaric acid production. The highest fumaric acid concentration (27.3 g/L) and yield (0.7 g/g of total consumed sugars) were achieved when the initial FAN concentration was 200 mg/L. The combination of VHP sugar with soybean cake hydrolysate derived from crude enzymes produced by SSF of A. oryzae at 200 mg/L initial FAN concentration led to the production of 40 g/L fumaric acid with a yield of 0.86 g/g of total consumed sugars. The utilization of sugarcane molasses led to low fumaric acid production by R. arrhizus, probably due to the presence of various minerals and phenolic compounds. The promising results achieved through the valorization of VHP sugar and soybean cake suggest that a focused study on molasses pretreatment could lead to enhanced fumaric acid production.     

城市生活垃圾中纤维素含量测定方法优化

对传统硫酸法进行改进，建立适用于城市生活垃圾纤维素含量的测定方法。以脂肪抽提时间、硫酸浓度、水解反应温度、水解反应时间、稀释硫酸浓度、回流时间为考察因素进行6因素5水平正交实验。结果表明，测定纤维素含量的最佳条件为：脂肪抽提时间4h、硫酸浓度70％、水解反应温度30℃、水解反应时间5h、稀释硫酸浓度5％、回流时间3h。优化后纤维素测定条件RSD均小于1．69％，平均回收率98．76％，表明该方法的精度高，可靠行强。     

Solvent tolerant marine bacterium Bacillus aquimaris secreting organic solvent stable alkaline cellulase

The organic solvent tolerant bacteria with their physiological abilities to decontaminate the organic pollutants have potentials to secrete extracellular enzymes of commercial importance. Of the 19 marine bacterial isolates examined for their solvent tolerance at 10 vol.% concentration, one had the significant tolerance and showed a relative growth yield of 86% for acetone, 71% for methanol, 52% for benzene, 35% for heptane, 24% for toluene and 19% for ethylacetate. The phylogenetic analysis of this strain using 16S rDNA sequence revealed 99% homology with Bacillus aquimaris. The cellulase enzyme secreted by this strain under normal conditions showed an optimum activity at pH 11 and 45 °C. The enzyme did show functional stability even at higher pH (12) and temperature (75 °C) with residual activity of 85% and 95% respectively. The enzyme activity in the presence of different additives were in the following order: Co⁺² > Fe⁺² > NaOCl₂ > CuSO₄ > KCl > NaCl. The enzyme stability in the presence of solvents at 20 vol.% concentration was highest in benzene with 122% followed by methanol (85%), acetone (75%), toluene (73%) and heptane (42%). The pre-incubation of enzyme in ionic liquids such as 1-ethyl-3-methylimidazolium methanesulfonate and 1-ethyl-3-methylimidazolium bromide increased its activity to 150% and 155% respectively. The change in fatty acid profile with different solvents further elucidated the physiological adaptations of the strain to tolerate such extreme conditions.     

剩余污泥水解酸化液磷去除的影响因素研究

城市污水厂剩余污泥水解酸化后可产生高浓度挥发性有机酸(VFAs),其中的乙酸和丙酸是增强生物除磷(EBPR)工艺的有利基质.但水解酸化液中含有大量的磷,如不进行处理就作为碳源回用到污水处理工艺中,势必增加除磷负荷.利用鸟粪石沉淀法可以去除污水中的磷.对城市污水厂剩余污泥水解酸化液形成鸟粪石的影响因素进行了试验研究.结果表明,在最佳工艺条件下,正磷和总磷的去除率分别可达92.5%和83.8%.