纤维素酶产生菌的选育及发酵条件优化

通过紫外线和化学诱变方法对一株纤维素酶产生菌--绿色木霉他进行了菌株选育,提高了产酶能力,变异株T211的CMCase和FPA分别达到19.17 IU/mL和1.94 IU/mL.进一步对其发酵条件进行研究,优化了培养基和培养条件.优化的培养基为:微晶纤维素10 g/L,麸皮10 g/L,蛋白胨0.4 g/L,尿素0.4 g/L,硫酸铵2.0 g/L,吐温-80 2.0 mL/L,其他同Mandel's营养盐液;培养条件包括温度、摇瓶装量、转速和接种量分别为:27～28℃、50 mL/250 mL、180 r/min和4%～6%.在上述条件下,CMCase达到25 IU/mL,FPA达到2.5 IU/mL.     

一株高产纤维素酶真菌的分离及产酶特性研究

一株高产纤维素酶真菌D1,经纤维素刚果红平板识别、滤纸崩解实验初筛以及液体发酵产酶复筛,从腐烂秸秆中被分离出来.经形态观察和18S rDNA基因片断分析,被鉴定为草酸青霉(Penicillium oxalicum).通过单因子及正交实验,D1液态发酵产酶条件得到优化.最佳产酶条件为:碳源为麸皮∶稻草=3∶7(w/w),...     

纤维素降解细菌筛选及降解特性分析

基于获得高效纤维素降解细菌的目的,通过LB培养基的培养以及刚果红培养基的筛选,从牛粪堆肥中筛选获得2株高效纤维素降解细菌。经鉴定,分别为枯草芽胞杆菌(Bacillus subtilis)和地衣芽胞杆菌(Bacillus licheniformis)。所筛选得到的菌种具有很高的滤纸降解能力,可在6d内使滤纸剧烈崩溃,振摇成均匀糊状;其中,地衣芽胞杆菌的羧甲基纤维素钠酶活峰值在发酵第4天达到峰值(237U/g)。     

碳氮比对猪粪和稻秆协同生物降解的影响

农业废弃物猪粪和稻秆产量巨大,因缺乏有效的处置方式,从而造成了巨大的污染,阻碍了现代农业的可持续发展。本研究以猪粪和稻秆的协同高效降解为研究导向,首次采用纤维素分解复合菌系预处理技术进行协同降解。通过考察碳氮比为10∶1、13∶1、16∶1和19∶1等4个梯度下的猪粪和稻秆混合物协同降解情况,并测定不同碳氮比不同培养时间下降解液的关键参数,探讨了不同碳氮比条件下复合菌系对农业废弃物协同降解效果的影响。结果表明,该菌系在碳氮比为16∶1、固形物含量为6%、接种量为10%的条件下于55℃下培养96 h,对猪粪和稻秆混合物的降解效果最佳。在此条件下其失重率高达26.2%,降解过程中的关键酶——滤纸酶和羧甲基纤维素酶(CMC酶)酶活可分别达到3.2和10.8 IU。该复合菌系在对猪粪和稻秆混合物协同降解过程中除了添加少许氯化钠外,无需再额外添加其他营养物质,展现出了良好的实际应用前景,可进一步为其他种类的农业废弃物的高效生物处置提供一定的理论依据。     

嗜麦芽寡养单胞菌DHHJ固态发酵羽毛工艺的研究

为了将微生物处理羽毛工艺产业化,利用课题组筛选得到的嗜麦芽寡养单胞菌,在液态发酵羽毛工艺的基础上进行固态发酵工艺的研究。通过发酵条件和培养基成分的优化,确定了嗜麦芽寡养单胞菌固态发酵羽毛的工艺为:初始pH为7.5,培养温度为35℃,培养时间为4 d,培养基组成是麸皮和干酪素,固液比是1∶1.5,照此工艺发酵,发酵液中蛋白质含量为23.78 mg/mL,角蛋白酶活为20.46 U/mL,菌株的产酶能力及降解羽毛能力显著高于液态发酵条件。     

一株木材腐朽菌及其粗酶液对七氯的降解

Phlebia acanthocystis TMIC34875是一株具有七氯降解能力的木材腐朽菌。为利用微生物技术去除环境中的七氯残留提供理论依据,研究了该菌株及其粗酶液对七氯的降解性能及其动力学特性。结果表明,菌株在七氯的初始浓度为50μmol/L时具有最大降解速率,为0.3031μmol/（L·h）;而菌体接种量为15%时,降解速率达到最高,为0.2045μmol/（L·h）。降解酶定位研究表明,七氯的降解主要是胞内酶在起作用。七氯胞内酶降解的酶促反应最适温度是35℃,在30-40℃之间有较高的催化活性;最适pH值为5.0,在pH 4.5-6.0之间有较高的催化活性,最适条件下反应1 h后七氯的降解率为65%。胞内粗酶液降解七氯的米氏常数K m为5.42μmol/L,最大反应速率V max为4.55μmol/min。胞内酶处理体系的GC/MS图谱显示,主要降解产物为1-羟基六氯、1-羟基-2,3-环氧六氯和环氧七氯,表明胞内酶对七氯的初始代谢机理同菌株相似,均是通过环氧化和置换反应来完成的。     

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。     

影响原油生物降解因素的实验研究

以原油为唯一碳源,从长期被石油污染土壤中筛选出6株原油降解菌SY1~SY6,通过单因素实验研究初始pH、温度、充氧量(摇床转数)、盐浓度、氮源和磷源等环境因素和营养条件对各菌株生物降解原油的影响。实验结果表明:6株原油降解菌在初始pH7~9,温度30℃,摇床转速180 r/min时生长良好,且能有效地降解石油类污染物,其平均降解率为50%以上。6菌株在盐浓度在1%时生长良好,SY3菌和SY4菌能在盐浓度10%以上生长,具有一定的耐盐能力。同时,6菌株以氯化铵(NH4Cl)为氮源,磷酸氢二钾(K2HPO4)和磷酸二氢钾(KH2PO4)的混合物(2∶1)为磷源时生长良好,因此可作为各菌生长的最适氮源和磷源。研究结果可以为含油废水的处理提供微生物基础。     

两步法处理含油污水的研究

利用微生物法,分絮凝和降解两步处理含油污水。第1步优化了高效产絮凝剂菌絮凝条件。结果表明,培养40h的产絮凝剂菌液具有最佳的絮凝活性,菌液质量浓度为0.25g/L,投加量为6 mL/L,pH为5~14,絮凝助剂为CaCl2时絮凝效果最佳。第2步优化了高效降解菌的降解条件。结果表明,以原油为碳源,鱼粉为氮源,碳氮比(质量比)20∶1,初始pH=8,含油率9.0%,含水率60%时,降解菌降解率最高。在以上优化条件下,含油污水的原油去除率可达77.4%。因此,全程利用微生物,分絮凝富集、固体培养两步无害高效处理含油污水是可行的。     

聚合物驱采出水中聚丙烯酰胺的微生物联合降解作用研究

通过对2株细菌的培养降解实验研究聚丙烯酰胺(hydrolyzed polyacrylamide,HPAM)降解菌对水环境下聚丙烯酰胺的降解作用,讨论协同降解机理。2株降解聚丙烯酰胺的菌株假单胞菌CJ419、枯草芽孢杆菌FA16在初始30℃废水样品上培养,定期测量细菌生物量和HPAM降解率。培养30 d后CJ419和FA16对聚合物的降解率最大值分别达到30.4%和25%,而以1∶1比例的混合菌降解率最大值达到80.3%。对2株菌胞外各组分研究表明:混合菌降解HPAM的机理主要由胞外降解酶系水解聚合物侧链基团导致HPAM降解为小分子物质,同时生长过程中降解菌还会释放非蛋白还原性物质引发氧化反应共同参与HPAM降解。     

Ultraviolet light-emitting diodes in water disinfection

Background, aim, and scope  The novel system of ultraviolet light-emitting diodes (UV LEDs) was studied in water disinfection. Conventional UV lamps, like mercury vapor lamp, consume much energy and are considered to be problem waste after use. UV LEDs are energy efficient and free of toxicants. This study showed the suitability of LEDs in disinfection and provided information of the effect of two emitted wavelengths and different test mediums to Escherichia coli destruction. Materials and methods  Common laboratory strain of E. coli (K12) was used and the effects of two emitted wavelengths (269 and 276 nm) were investigated with two photolytic batch reactors both including ten LEDs. The effects of test medium were examined with ultrapure water, nutrient and water, and nutrient and water with humic acids. Results  Efficiency of reactors was almost the same even though the one emitting higher wavelength had doubled optical power compared to the other. Therefore, the effect of wavelength was evident and the radiation emitted at 269 nm was more powerful. Also, the impact of background was studied and noticed to have only slight deteriorating effect. In the 5-min experiment, the bacterial reduction of three to four log colony-forming units (CFU) per cubic centimeter was achieved, in all cases. Discussion  When turbidity of the test medium was greater, part of the UV radiation was spent on the absorption and reactions with extra substances on liquid. Humic acids can also coat the bacteria reducing the sensitivity of the cells to UV light. The lower wavelength was distinctly more efficient when the optical power is considered, even though the difference of wavelengths was small. The reason presumably is the greater absorption of DNA causing more efficient bacterial breakage. Conclusions  UV LEDs were efficient in E. coli destruction, even if LEDs were considered to have rather low optical power. The effect of wavelengths was noticeable but the test medium did not have much impact. Recommendations and perspectives  This study found UV LEDs to be an optimal method for bacterial disinfection. The emitted wavelength was found to be an essential factor when using LEDs; thus, care should be taken in selecting the proper LED for maximum disinfection.     

Bioaugmentation with butane-utilizing microorganisms to promote in situ cometabolic treatment of 1,1,1-trichloroethane and 1,1-dichloroethene

A field study was performed to evaluate the potential for in-situ aerobic cometabolism of 1,1,1-trichloroethane (1,1,1-TCA) through bioaugmentation with a butane enrichment culture containing predominantly two Rhodococcus sp. strains named 179BP and 183BP that could cometabolize 1,1,1-TCA and 1,1-dicholoroethene (1,1-DCE). Batch tests indicated that 1,1-DCE was more rapidly transformed than 1,1,1-TCA by both strains with 183BP being the most effective organism. This second in a series of bioaugmentation field studies was conducted in the saturated zone at the Moffett Field In Situ Test Facility in California. In the previous test, bioaugmentation with an enrichment culture containing the 183BP strain achieved short term in situ treatment of 1,1-DCE, 1,1,1-TCA, and 1,1-dichloroethane (1,1-DCA). However, transformation activity towards 1,1,1-TCA was lost over the course of the study. The goal of this second study was to determine if more effective and long-term treatment of 1,1,1-TCA could be achieved through bioaugmentation with a highly enriched culture containing 179BP and 183BP strains. Upon bioaugmentation and continuous addition of butane and dissolved oxygen and or hydrogen peroxide as sources of dissolved oxygen, about 70% removal of 1,1,1-TCA was initially achieved. 1,1-DCE that was present as a trace contaminant was also effectively removed ( 80%). No removal of 1,1,1-TCA resulted in a control test leg that was not bioaugmented, although butane and oxygen consumption by the indigenous populations was similar to that in the bioaugmented test leg. However, with prolonged treatment, removal of 1,1,1-TCA in the bioaugmented leg decreased to about 50 to 60%. Hydrogen pexoxide (H₂O₂) injection increased dissolved oxygen concentration, thus permitting more butane addition into the test zone, but more effective 1,1,1-TCA treatment did not result. The results showed bioaugmentation with the enrichment cultures was effective in enhancing the cometabolic treatment of 1,1,1-TCA and low concentrations of 1,1-DCE over the entire period of the 50-day test. Compared to the first season of testing, cometabolic treatment of 1,1,1-TCA was not lost. The better performance achieved in the second season of testing may be attributed to less 1,1-DCE transformation product toxicity, more effective addition of butane, and bioaugmentation with the highly enriched dual culture.     

一株高效脱硫菌的分离鉴定和脱硫特性研究

从污水净化厂二沉池回流污泥中分离到一株硫杆菌菌株dj-5，该菌株是一种兼性厌氧菌,通过生理生化特性鉴定,并结合16S rDNA序列分析及鉴定,该菌株可以确定为脱氮硫杆菌（Thiobacillus denitrificans）。该菌株的生长曲线表明菌体生长迅速,延滞期约为8 h,然后细菌进入对数生长期,这一阶段持续时间大约20～24 h,稳定期较短,细菌很快进入衰亡期。通过脱硫工艺实验考察了pH值、温度和搅拌速度对脱硫率和菌体生长的影响,结果表明，在进气量180 L/h、pH=6～8、温度为25～35℃和搅拌速度为270 r/min时,该菌株能正常生长,对进气浓度高达2 500 mg/m³的硫化氢脱除率在91%以上。     

脱氮副球菌YF1微生物燃料电池生物阴极脱氮和产电

以脱氮副球菌YF1构建纯种生物阴极微生物燃料电池(microbial fuel cell,MFC)进行脱氮和产电机理的研究。研究结果发现,阴极碳氮比、pH值对产电和脱氮效率有明显影响。当MFC的阴极运行条件pH值为8.0,碳氮比为20时,运行时间15 h时,脱氮率高达100%,输出电压为150 mV。上述结果表明,微生物燃料电池运行过程中,细菌降解硝酸根的机理为将硝酸根还原为N2或者直接将其作为自身的营养物质而利用。循环伏安(CV)与扫描电镜(SEM)的结果表明,在微生物燃料电池运行中,副球菌YF1通过接触导电作为产电的电子供体。     

对氨基苯磺酸降解菌的分离及其特性研究

从温州地区受污染的河水中分离到一株能降解对氨基苯磺酸的菌株WZR-3，该菌株能以对氨基苯磺酸为惟一碳源、能源生长。经对其形态特征、生理生化以及16S rDNA序列分析，该菌株初步鉴定为人苍白杆菌(Ochrobactrum anthropi)。该菌株利用对氨基苯磺酸生长时最适生长温度和pH值分别为30℃和7。该菌在10 g/L对氨基苯磺酸时仍能生长，最适生长浓度为300 mg/L对氨基苯磺酸。降解底物广谱性测试表明，该菌株还能降解多种芳香类化合物。     

城市生活污水中志贺氏菌ipaH毒力基因的定量PCR检测

基于ipaH毒力基因的实时荧光定量PCR检测,建立适合城市污水中志贺氏菌的定量检测方法。使用从临床分离出的志贺氏菌构建重组质粒作为实时荧光定量PCR的标准品。在ipaH基因模板量2.58×100～2.58×106copy范围内具有良好的线性关系,每100 mL水样中含有2.58×101copy以上的ipaH基因即可被检出。从西安市生活污水分离得到2株野生型志贺氏菌,分析ipaH基因数量和菌体数量的关系,从而确定ipaH基因定量检测志贺氏菌的可行性。该方法灵敏、快速、特异性好,适用于城市生活污水中志贺氏菌的检测。     

汞、铅、铬污染土壤的微生物修复

利用裂褶菌(Schizophyllum commune)GGHN08-116菌株,以棉籽壳、玉米秸等为固体发酵底物修复受汞、铅、铬污染的土壤。通过菌丝穿透重度重金属土壤实验,研究了菌丝在穿透土壤过程对交换态重金属的影响以及该菌株子实体对重金属离子的富集能力,同时,通过盆栽实验研究了在重度重金属污染土壤上,施用不同比例的固体发酵料对污染土壤中汞、铅、铬及其胡萝卜根茎质量、产量的影响,研究结果表明,该菌株能穿透厚度为5 cm的土壤,并有子实体生成,土壤pH值略有下降,与对照差异不显著;与对照相比,土壤中交换态汞、铬含量均显著下降,而交换态铅差异不显著,子实体中除汞含量符合标准外,铅、铬均超出了GB 7096-2003,GB 2762-2005规定标准。在固体发酵料处理下土壤中交换态汞、铅、铬含量均显著下降,胡萝卜根茎中均未检测到汞、铅含量,铬含量也符合GB 2762-2005规定标准。GGHN08-116菌株及其固体发酵产物具有修复受重金属污染土壤的能力。     

高效脱硫菌的筛选及其性能研究

从高硫污染的活性污泥中富集培养，分离纯化得到一株可以降解噻吩的菌株s_4，并对该菌株的形态特征进行观察。应用Design—Expert8．0．5b软件进行响应面优化实验，研究了反应时间、噻吩浓度、微生物浓度3个因素的组合对菌株s_4脱硫效果的影响，并拟合得到多元二次回归方程，得出最佳实验条件。拟合结果表明，当反应时间27．46h，噻吩浓度为1．04％，微生物浓度4．04％时，预测噻吩降解率为14．8％，通过验证得最佳条件下的降解率为14．3％，与预测值相符。     

高效脱硫菌的筛选及其性能研究简

从高硫污染的活性污泥中富集培养，分离纯化得到一株可以降解噻吩的菌株S-4，并对该菌株的形态特征进行观察。应用Design-Expert8.0.5b软件进行响应面优化实验，研究了反应时间、噻吩浓度、微生物浓度3个因素的组合对菌株S-4脱硫效果的影响，并拟合得到多元二次回归方程，得出最佳实验条件。拟合结果表明，当反应时间27.46 h，噻吩浓度为1.04%，微生物浓度4.04%时，预测噻吩降解率为14.8%，通过验证得最佳条件下的降解率为14.3%，与预测值相符。