Characterization of the effective cellulose degrading strain CTL-6

An efficient cellulose degrading bacteria exists in the thermophilic wheat straw-degrading community, WDC2. However, this strain cannot be isolated and cultured using conventional separation techniques under strict anaerobic conditions. We successfully isolated a strain of effective cellulose degrading bacteria CTL-6 using a wash, heat shock, and solid-liquid alternating process. Analysis of its properties revealed that, although the community containing the strain CTL-6 grew under aerobic conditions, the purified strain CTL-6 only grew under anaerobic culture conditions. The strain CTL-6 had a striking capability of degrading cellulose (80.9% weight loss after 9 days of culture). The highest efficiency value of the endocellulase (CMCase activity) was 0.404 μtmol/(min-mL), cellulose degradation efficiency by CTL-6 was remarkably high at 50-65°C with the highest degradation efficiency observed at 60°C. The 16S rRNA gene sequence analysis indicated that the closest relative to strain CTL-6 belonged to the genus Clostridium thermocellum. Strain CTL-6 was capable of utilizing cellulose, celiobiose, and glucose. Strain CTL-6 also grew with Sorbitol as the sole carbon source, whereas C.therrnocellum is unable to do so.     

一株高效降解菲的植物内生细菌筛选及其生长特性

从长期受PAHs污染的植物看麦娘中分离出1株可高效降解菲的内生细菌Pn2,经生理生化特征分析和16S rDNA序列同源性分析,初步鉴定为Naxibacter sp.,研究了菌株Pn2的生长特性及其对菲的降解作用.结果表明,菌株Pn2能以菲为唯一碳源生长,并对菲有良好的降解性能.菲浓度为49.92mg/L时,30℃下150r/min振荡培养72h,菲降解率高达98.78%.接种量和污染强度显著影响Pn2对菲的降解:接种量越大,菲降解率越高;随污染强度升高,菲降解率先增大后减小,最适污染强度为150mg/L.菌株Pn2有较强的环境适应能力.温度为25~37℃、环境pH值为6.0~8.0、盐浓度1%~2%范围内,菌株Pn2生长状况良好.菌株Pn2为好氧生长,通气量越大,菌株生长越旺盛.菌株Pn2对低浓度的氨苄青霉素和氯霉素有抗性.     

一株发酵纤维素产丁酸菌及其代谢动力学特性

为开发降解纤维素产丁酸菌的种子资源,从牛粪、猪粪堆肥、玉米地土壤和腐木混合物的富集样品中分离得到一株厌氧降解纤维素产丁酸菌.该菌株细胞呈杆状,长7.1~9.1μm,直径1.2μm左右,经鉴定为丁酸梭菌(Clostridium butyricum),命名为C. Butyricum DCB.在35℃条件下,菌株DCB在纤维二糖液体培养基中的最大比生长速率为0.6536h^-1,世代时间为1.06h,纤维二糖降解速率为0.1g/(L·h),丁酸生成速率为0.06g/(L·h).该菌株利用纤维素发酵产丁酸的转化率高达0.23g/g,具有良好的开发前景.     

多功能细菌复合系NSC-7的菌种组成多样性

NSC-7是一组具有降解纤维素和林丹双重功能的细菌复合系.为系统了解复合系的菌种组成,在有氧条件下,利用传统的平板画线法分离到11株单菌,将11株单菌按体积比1∶1重新组合并不具备分解纤维素的能力,利用单层和双层平板滤纸法检测NSC-7的分解能力,发现只有双层平板上的滤纸变黄且降解,说明复合系内纤维素降解的关键菌是厌氧或微好氧菌.利用现代分子生物学技术对NSC-7构建克隆文库,获得了195个16S rDNA片断,经DGGE筛选获得25个代表克隆,其序列数据库比对结果中有60％的近缘种为已知菌,分别归属于Clostridium、Petrobacter、Bacteria、Paenibacillus、Proteobacterium 5个属,其余40％的近缘种为难培养菌株.     

辛硫磷降解菌XSP-1的分离、鉴定及其降解特性研究

从农药厂污泥中分离到1株能以辛硫磷为唯一碳源生长的细菌, 命名为XSP-1.根据其生理生化特征和16S rRNA基因序列相似性分析, 将该菌株初步鉴定为戴尔福特菌属(Delftia sp.). 该菌株能在7 h内完全降解100　mg/L的辛硫磷. XSP-1降解辛硫磷的最适pH为7.0, 最适温度为35℃, 降解速率与初始接种量呈正相关, 该菌株对甲基对硫磷、毒死蜱、杀螟硫磷也有较好的降解能力. 根据已报道有机磷农药降解基因mpd的保守序列设计引物, 未从该菌株中扩增到目标条带, 但是该菌株是否带有新的有机磷农药降解基因仍需要进一步研究.     

毒死蜱降解细菌XZ-3的分离及降解特性研究

从某农药厂排污口的污泥中采样,经驯化富集后筛选到1株能高效降解毒死蜱的细菌XZ-3,经过对其形态、生理生化特征及16S rDNA的分析,鉴定该菌株为节杆菌属（Arthrobactersp.）.测定了不同碳源、pH、温度及毒死蜱浓度对细菌降解能力和生长量的影响,以培养液在波长400 nm下的光密度值表示细菌生长量,液体介质中毒死蜱的提取采用漩涡振荡提取法,提取溶剂为石油醚,毒死蜱的测定采用紫外分光光度法.结果表明,该菌株24 h内对100 mg·L^-1毒死蜱的降解率高达86.8%;在外加碳源浓度为0.3%时降解率最大;细菌的生长量随着外加碳源浓度的升高而增加;pH在偏酸和偏碱性的条件下降解率较大,pH 9.0时达到最大,细菌的生长量在pH 8.0～10.0偏碱性的条件下较大;在毒死蜱浓度为100　mg·L^-1时降解率最大,该菌具有较强的抗药性,当毒死蜱浓度达到1?000　mg·L^-1时仍能生长,细菌的生长量在800　mg·L^-1时达到最大,绝对去除量随毒死蜱浓度的提高而增加;细菌的生长和降解需要适宜的温度,30℃培养时,降解率和生长量最大.本研究提出了细菌XZ-3生长和降解毒死蜱的最佳培养条件,可为农药污染治理及生产污水处理提供理论依据.     

丁二腈高效降解菌的筛选及其降解性

以丁二腈为唯一碳源和氮源,从石化腈纶废水及其处理构筑物的生物膜中,分离、筛选出2株高效降解丁二腈的菌株:J-1-3和J-13-1.经形态学观察和生理生化特征研究,两者均被鉴定为假单胞菌(Pseudomonas spp.).通过摇瓶试验得出2菌株的最适生长条件:温度为30℃,摇床转速(间接反映通气量)为250 r/min,接种量为0.1%,初始pH为6.在最适生长条件下,分别对不同初始浓度丁二腈进行降解率试验.结果表明,2菌株对丁二腈的降解能力强,尤以J-13-1更为显著.当丁二腈的初始浓度约为6000mg/L、8000mg/L和10000mg/L时,J-13-1菌株对丁二腈的降解率分别在12.5h、14h和16h时达到100%.     

耐盐石油烃降解菌的筛选鉴定及其特性研究

为得到高效耐盐石油烃降解菌,从黄河三角洲石油污染盐渍化土壤中分离出39株细菌,经液体培养初筛和土壤培养复筛实验,得到1株高效耐盐石油烃降解菌BM38.通过形态特征、生理生化特征和16S rDNA序列分析,确定该菌为恶臭假单胞菌（Pseudomonas putida）.通过液体培养实验,研究了BM38的耐盐和产生物表面活性剂特性以及对不同烃的利用能力.结果表明,在含0.5%～6.0%NaCl液体培养基中BM38生长良好,属中度耐盐菌.在高盐环境下BM38具有较强的分解石油烃能力,其中在含1.0%NaCl液体培养基中,降解7 d后,原油降解率达到73.5%;在含盐量0.22%和0.61%土壤中添加BM38,降解40 d后,土壤总石油烃降解率达到40%以上.BM38能产生一种生物乳化剂,盐浓度对这种乳化剂的乳化能力影响较大,当NaCl浓度增加到1.0%,乳化值（EI24）开始迅速降低,但在NaCl浓度为2.0%时,EI24仍达到61.0%.BM38能够利用环己烷、甲苯、异辛烷、菲和正十六烷为唯一碳源生长,其中对正构烷烃和芳烃具有较强的利用能力.     

汽油降解菌的分离及降解研究

从泄漏污染的加油站土壤中筛选出对汽油具有较强降解能力的菌株,研究该菌株最适宜的生长条件,探讨紫外线诱导及投加表面活性剂等强化手段对该菌株降解汽油的影响.结果表明:①通过富集培养的方法分离得到的菌株Q18,经形态特征及生理生化特征鉴定,初步确定其为红球菌(Rhodococcus sp.).②菌株Q18在培养液中适宜生长的温度,pH和底物质量浓度分别为35 ℃,6.0和1 000 mg/L.③通过紫外线照射诱变后的菌株降解能力强于原始菌株,且15 W紫外灯对菌株的诱变效果优于30 W;氯化锂单独诱变效果不明显;经紫外灯照射和氯化锂复合诱变的菌株QY4对汽油的降解率达到了52.2%,在所有诱变菌中最高,效果最显著. ④表面活性剂能增强汽油的生物可利用性,强化菌株Q18对汽油的降解,但阴离子和非离子的混合表面活性剂SDS+TX-100和SDS+TW-80比单一表面活性剂更能有效提高菌株Q18对汽油的降解率.      

Degradation of 2,6-di-tert-butylphenol by an isolated high-efficiency bacterium strain

An aerobic bacterium strain, F-3-4, capable of effectively degrading 2,6-di-tert-butylphenol(2,6-DTBP), was isolated and screened out from an acrylic fiber wastewater and the biofilm in the wastewater treatment facilities. This strain was identified as Alcaligenes sp. through morphological, physiological and biochemical examinations. After cultivation, the strain was enhanced by 26.3% in its degradation capacity for 2,6-DTBP. Results indicated that the strain was able to utilize 2,6-DTBP, lysine, lactamine, citrate, n-utenedioic acid and malic acid as the sole carbon and energy source, alkalinize acetamide, asparagine, L-histidine, acetate, citrate and propionate,but failed to utilize glucose, D-fructose, D-seminose, D-xylose, sedne and phenylalanine as the sole carbon and energy source. The optimal growth conditions were determined to be: temperature 37℃, pH 7.0, inoculum size 0.1% and shaker rotary speed 250 r/min. Under the optimal conditions, the degradation kinetics of 2,6-DTBP with an initial concentration of 100 mg/L was studied. Results indicated that 62.4% of 2,6-DTBP was removed after 11 d. The degradation kinetics could be expressed by Eckenfelder equation with a half life of 9.38 d. In addition, the initial concentration of 2,6-DTBP played an important role on the degradation ability of the strain. The maximum initial concentration of 2,6-DTBP was determined to be 200 mg/L. Above this level, the strain was overloaded and exhibited significant inhibition.     

复合菌系BYND-8的种群组成及其对沼气产量的影响

为了明确1组在中温下(30℃)高效分解木质纤维素的复合菌系的菌群组成.研究复合菌系预处理秸秆对沼气发酵的影响,利用平板分离法和变性梯度凝胶电泳(DGGE)法研究了中温木质纤维素降解复合菌系BYND-8的菌种组成多样性,通过添加该复合系菌液到以牛粪为原料的沼气发酵体系,研究了添加秸秆降解液对沼气产量的影响.利用平板法分离...     

降解纤维素产甲烷的四菌复合系

自然环境中通常是微生物群落协同完成纤维素的降解,构建可降解纤维素的混菌体系是认识微生物相互作用的关键.利用富集培养法,结合变性梯度凝胶电泳(DGGE)指纹检测技术以及厌氧滚管技术,建立了一种筛选简单降解纤维素产甲烷复合菌系的方法.利用此方法从青藏高原若尔盖高寒湿地分离到一个由4株菌构成的降解纤维素产甲烷的稳定菌系.结果表明,该复合菌系由纤维素水解菌Clostridium glycolicum、非纤维素水解菌Trichococcus flocculiformis和Parabacteroides merdae、产甲烷古菌Methanobacterium subterraneum等具有不同功能的4种菌株组成,且在这4株菌的共同作用下可将纤维素直接转化为CH4.该简单复合系的获得为今后纤维素转化甲烷复合菌系的代谢控制和遗传改造提供了一个平台.     

咪唑乙烟酸降解菌的分离、鉴定及其降解特性研究

从生产咪唑乙烟酸化工厂排污口的污泥和长期施用咪唑乙烟酸的混合土壤中分离到1株能降解咪唑乙烟酸的细菌.该菌株在72h内对500mS/L的咪唑乙烟酸降解率达到90%以上. pU为5时,500mS/L的咪唑乙烟酸72h内可全部降解,而pU 8和pH9条件下,72h咪唑乙烟酸的降解率仅为50%左右,酸性条件比碱性条件更适合降解菌的生长.25℃和30℃条件下,降解菌对咪唑乙烟酸的降解效率较高.25℃, pH 5是降解菌对咪唑乙烟酸降解的最佳条件.从形态特征、生理生化特性及 16S rRNA序列分析鉴定该菌株属于产碱菌属.     

Process of rice straw degradation and dynamic trend of pH by the microbial community MC1

The process of the rice straw degradation in the fermentor with aeration at 290 ml/h was studied. The results of dissolved oxygen （DO） indicated that the optimum DO during cellulose degradation by microbial community MC1 ranged from 0.01 to 0.12 mg/L. The change model ofpH values was as follows： irrespective of the initial pH of the medium, pH values decreased rapidly to approximate 6.0 after being inoculated within 48 h when cellulose was strongly degraded, and then increased slowly to 8.0--9.0 until cellulose was degraded completely. During the degradation process, 15 kinds of organic compounds were checked out by GC-MS. Most of them were organic acids. Quantity analysis was carried out, and the maximum content compound was ethyl acetate which reached 13.56 g/L on the day 4. The cellulose degradation quantity and ratio analyses showed that less quantity （under batch fermentation conditions） and longer interval （under semi-fermentation conditions） of rice straw added to fermentation system were contributed to matching the change model of pH, and increasing the quantity and ratio of rice straw degradation during cellulose degrading process. The highest degradation ratio was observed under the condition office straw added one time every five days （under semi-fermentation conditions）.     

产碱菌株F-3-4对苯酚降解特性的研究

从腈纶废水处理构筑物中分离筛选到1株高效降解2,6-二叔丁基苯酚的菌株F-3-4,经驯化发现其对苯酚也有较好的降解能力。通过紫外吸收测定菌液生长值以及安替比林比色法测定苯酚浓度,考察了不同底物浓度、pH值、通气量对苯酚降解的影响以及苯酚降解的动力学分析。结果表明,该菌生长的迟滞期随苯酚浓度的增大而延长,苯酚浓度增大对菌株有明显的抑制作用。200mg/L苯酚浓度的完全降解时间在36h之内,该菌株降解苯酚基本发生在对数期,其对苯酚降解适宜条件为温度35℃,pH7～8,该菌为好氧菌,通气有利于苯酚的降解。在最适条件下其降解苯酚的动力学特征符合Monod模型。     

紫色非硫细菌的培养及处理酿酒废水的研究

从酿酒废水的活性污泥中分离、培养得到纯化的紫色非硫细菌01S菌株,并用其处理高浓度有机酿酒废水。结果表明,紫色非硫细菌在自然(或白炽灯)光照、pH值7.0、温度为28～30℃条件下生长良好,且能够明显降解酿酒废水,其COD的去除率达到82.2%左右。并探讨了该菌在两种不同条件下(光照厌氧和好氧黑暗)的降解效果,及其与DO值的关系。     

油烟污染物降解优势菌株的筛选及性能研究

在完成对活性污泥驯化的基础上,分离、纯化出XJ9,XJ30,XJ2,XJ21,XJ28,XJ25等6株油烟污染物降解细菌,研究其生长条件及降解性能,筛选出一株优势菌株并进行菌落形态学及分子生物学鉴定。实验结果表明:6株细菌最适生长温度在35℃左右、最适pH在6⁸之间、最佳摇床转速介于808之间、最佳摇床转速介于80¹60 r/min之间,各菌株最佳降解时间均为36 h,对油烟污染物的降解率介于12%160 r/min之间,各菌株最佳降解时间均为36 h,对油烟污染物的降解率介于12%⁶0%之间,其中XJ25最高可达到60%左右,可确定XJ25为油烟污染物降解优势菌株。经分子生物学鉴定,确定XJ25为干燥棒状杆菌(Corynebacterium xerosis)。所获得的油烟污染物降解菌对于应用生物法降解餐饮油烟污染物具有一定的价值。     

Biodegradation of nicosulfuron by a novel Alcaligenes faecalis strain ZWS11

A bacterial strain ZWS11 was isolated from sulfonylurea herbicide-contaminated farmland soil and identified as a potential nicosulfuron-degrading bacterium. Based on morphological and physicochemical characterization of the bacterium and phylogenetic analysis of the 16S rRNA sequence, strain ZWS11 was identified as Alcaligenes faecalis. The effects of the initial concentration of nicosulfuron, inoculation volume, and medium pH on degradation of nicosulfuron were investigated. Strain ZWS11 could degrade 80.56% of the initial nicosulfuron supplemented at 500.0 mg/L under the conditions of pH 7.0, 180 r/min and 30°C after incubation for 6 days. Strain ZWS11 was also capable of degrading rimsulfuron, tribenuron-methyl and thifensulfuron-methyl. Four metabolites from biodegradation of nicosulfuron were identified, which were 2-aminosulfonyl-N, N-dimethylnicotinamide (M1), 4, 6-dihydroxypyrimidine (M2), 2-amino-4, 6-dimethoxypyrimidine (M3) and 2-(1-(4,6-dimethoxy-pyrimidin-2-yl)-ureido)-N,N-dimethyl-nicotinamide (M4). Among the metabolites detected, M2 was reported for the first time. Possible biodegradation pathways of nicosulfuron by strain ZWS11 were proposed. The degradation proceeded mainly via cleavage of the sulfonylurea bridge, O-dealkylation, and contraction of the sulfonylurea bridge by elimination of a sulfur dioxide group. The results provide valuable information for degradation of nicosulfuron in contaminated environments.     

焦化废水中苯酚降解菌筛选及其降解性能

焦化废水中苯酚类及其衍生物的降解率高低是焦化废水COD是否达标排放的关键.采用不同培养基和菌种驯化方法,从焦化废水厂活性污泥中分离筛选获得4株苯酚降解菌,经生理生化和16S rDNA分子鉴定,A1为球杆菌属Sphaerobacter,C1为鲍曼不动杆菌Acinetobacter baumannii;D2为睾丸酮丛毛单胞菌Comamonas testosterone;D3为Novosphingobiumnaphthalenivorans.4株降酚菌均具有较高的苯酚耐受力和降解效率,是生物法处理酚类污染废水优质的种质资源.菌株D2不仅对苯酚具较高耐受力达到2 000 mg.L-1、且在48 h内可将初始浓度为1 000 mg.L-1的苯酚完全降解.环境因子考察研究表明,pH为7.5～8.5,温度为30～40℃范围内,转速为150 r.min-1,是菌株D2的最优降解条件,本研究结果为构建高效处理焦化废水基因工程菌提供了微生物基础.     

苯酚微生物降解的正交实验研究

利用以苯酚为碳源的驯化液,对某焦化厂曝气池活性污泥进行驯化,经过分离、筛选,挑选出4株高效的苯酚降解菌,编号为h32a2、b31B、h31A和b41a,并通过菌落形态特征、简单染色及生理生化反应初步鉴定为假单胞菌属(Pseudomonas sp.),依据正交实验确定了优势菌群降解苯酚的最佳条件为温度32℃,pH值7.5,菌培养时间16 h,接种量1%。