一株菲降解菌的分离鉴定及其降解条件研究

以菲为研究对象,从克拉玛依稠油污染土壤中筛选到1株对菲具有较好降解效果的菌株JZ3-21。通过形态观察、生理生化指标及16S rDNA序列分析对该菌株进行了鉴定。该株菌的16S rDNA序列与Pseudomonas属的相似性达99%,结合分离菌株的形态、生理生化特征和16S rDNA基因序列的分析结果,初步鉴定该菌株为恶臭假单胞菌（Pseudomonas putida.）。对其降解条件进行了研究,结果表明：在40℃,pH 8.0,接种量为1.5%的条件下,菌株对初始质量浓度为100 mg.L-1的菲在64 h内的降解率高达94.2%。该菌对高质量浓度菲有较好的耐受性,其最高耐受质量浓度可达2 000 mg.L-1。     

菲在硅酸盐矿物上的吸附与光化学降解行为

选用高岭石和蒙脱石为对象研究次生硅酸盐矿物对菲光降解过程产生的影响,并以石英砂作为对照.由于3种矿物的表面积、结构组成、表面电荷、阳离子交换能力不同,导致吸附性能不同,对菲的吸附能力呈现出蒙脱石高岭石石英砂.以发射波长为254 nm的紫外灯为光源,在3种矿物中进行菲光化学降解实验,探讨不同矿物中菲光化学降解行为的区别.紫外光辐射对3种矿物中菲的去除都符合一级反应动力学,24 h后菲的去除效果为:石英砂高岭石蒙脱石,与矿物吸附能力的大小有密切关系.与避光对照组相比,去除率整体上呈现出光照避光,表明紫外光辐射可以显著促进自然环境中菲的转化和降解.通过GC-MS检测到菲光降解的主要产物为邻苯二甲酸二(2-乙基己基)酯、烷烃、苯酚等,表明菲经过紫外光辐射会逐渐变成低毒或无毒物质,最终被矿化.     

蜡状芽孢杆菌不同菌株降解有机磷农药的特性分析

筛选分离降解微生物解决有机磷农药残留给水体和土壤环境带来的污染问题是一项可行的生物修复技术。采用富集培养和定时取样分析有机磷农药残留的方法,分离驯化出三株能够降解有机磷农药的细菌,研究了其形态特征和生理生化特性并对其16SrDNA序列进行了分析,同时比较了三菌株对甲基对硫磷（Methyl－parathion）、毒死蜱（Chlorpyrifos）和_二唑磷（Triazophos）的降解特性。结果表明：通过富集培养得到10菌株具有降解甲基对硫磷和毒死蜱的能力,比较确定HY-1、HY-2和HY-4三菌株作为研究对象,经鉴定为蜡状芽孢杆菌（Bacilluscereus）的不同菌株,三菌株在Genbank上的登录号分别为：eu915687、eu915686和eu915688。在甲基对硫磷质量浓度为50mg·L-1时,三菌株72h的降解率分别为91．7％、87．7％与92．4％．降解率无显著性差异（P〉0．05）,当甲基对硫磷质量浓度增加到100mg·L-1时,三菌株对甲基对硫磷的降解率有所下降,其中HY-2对甲基对硫磷的降解率下降最大达23％,且和其他两菌株有显著性差异（P〈0．05o三菌株72h对100mg·L-1毒死蜱的降解率分别达到64．8％、53．7％和56．5％,在不同的毒死蜱初始质量浓度下,HY-1和HY-4两菌株对毒死蜱的降解率无显著性差异（P〉0．05）,HY-2与HY-1、HY-4两菌株有显著性差异（P〈0．05o三菌株对三唑磷的降解率均较低,其中HY-2对初始质量浓度为100mg·L-1三唑磷的降解率最高仅为20．7％,其余两菌株对三唑磷的降解率比HY-2低且无显著性差异（P〉0．05o可以得出本研究分离得到的蜡状芽孢杆菌不同菌株对有机磷农药的降解存在多态性。     

乐果降解菌LGX1的筛选及其降解特性研究

通过富集培养,从连续施用农药乐果的土壤中分离得到一株具有较强降解有机磷农药乐果能力的细菌菌株LGX1,通过菌落形态观察及细菌的16SrDNA测序,对其进行了鉴定,同时初步研究了其降解性能。结果表明：该菌株为蜡状芽孢杆菌（Bacillus cereus）,在接种量为20%,乐果初始质量浓度为100mg·L-1,外加碳源葡萄糖质量分数为2%,温度为35℃,初始pH值为4时该菌株对乐果的降解能力最强;并且能在以辛硫磷、毒死蜱和三唑磷为唯一碳源的基础盐培养基中生长,且均比在以乐果为唯一碳源的基础盐培养基中生长的OD600值要高,初步推断菌株LGX1对有机磷农药的降解有一定的广谱性。     

一株菲降解菌的特性及相关降解基因的克隆

从石油污染土壤中分离到一株菲降解菌2F5-2.根据该菌株生理生化特征和16S rDNA序列相似性分析,将其初步鉴定为鞘氨醇杆菌属(Sphingobium sp.).该菌株在10 h内对100 mg/L的菲的降解率为100%.降解菲的最适温度为30℃,最适pH为7.对降解途径的初步研究显示,该菌株通过水杨酸途径降解菲.克隆了编码芳香烃双加氧酶α亚基的基因phdA,它与菌株Sphingomonas sp.P2、Sphingobium yanoikuyae B1、Sphingomonas sp.ZP1中phdA的同源性分别为97.9%、98%和100%,表明该基因具有保守性.图6参16     

一株高效邻苯二甲酸二丁酯降解菌的筛选、鉴定及其降解特性研究

从活性污泥中分离出1株以邻苯二甲酸二丁酯(DBP)为碳源和能源生长的高效降解菌DP-2,经形态观察、生化鉴定及16S rDNA序列分析,鉴定该菌株为不动杆菌(Acinetobacter sp.).采用单因素试验研究了不同试验条件(接种量、DBP浓度、NaCl浓度和碳源)对菌株DBP降解特性的影响,结果表明:接种量大于10%时,菌株DP-2在3 d内对初始浓度为10 mg·L~(-1)的DBP降解率可达到90%以上;DBP初始浓度为5—50 mg·L~(-1)时,菌株在6 d内对DBP降解率均能达到90%以上,但高浓度DBP会影响菌株DP-2生长,DBP浓度为1000 mg·L~(-1)时,DBP降解率仅为26.88%;菌株降解DBP的最佳NaCl浓度范围为0—20 g·L~(-1);此外,醋酸钠、蔗糖、葡萄糖添加对于菌株降解DBP均有一定的促进作用,其中葡萄糖效果最为明显.在此基础上,采用响应曲面法优化了菌株降解DBP的培养条件并进行了试验验证,在盐度为5 g·L~(-1),接种量为17.14%,底物浓度为9.81 mg·L~(-1),菌株对DBP的降解率为85.86%.     

一株高效甲醛降解菌的分离筛选与降解特性研究

甲醛是一种被广泛使用的重要化工原材料和有机溶剂,其35%~40%的水溶液是医学上和科研上常用的防腐剂。然而,甲醛作为一种原生毒素,具有强烈的致癌作用,将其释放到环境中不仅严重危害人体健康,而且具有极大的环境风险。利用微生物降解甲醛已成为治理甲醛污染的重要方法。为了获得高效的甲醛降解微生物,该研究采集北京某污水处理厂的活性污泥作为菌种分离源,利用稀释平板涂布及平板划线的方法分离纯化得到一株能以甲醛为唯一碳源生长的降解菌株,并将其命名为MCA01(CGMCC11443)。通过对该菌株的菌落形态和菌株形态进行观察,并采用法国生物-梅里埃公司的VITEK 2COMPACT全自动微生物分析系统及其配套的革兰氏阴性鉴定板对该菌株进行常规快速鉴定,结果显示,MCA01与恶臭假单胞菌(Pseudomonas putida)的相似度为99%。利用PCR扩增得到菌株的16S rRNA基因,并基于测序所得序列对菌株MCA01进行了BLAST比对及系统发育分析,结果指出此分离菌株与Pseudomonas putida的同源性为100%。综合上述方法,鉴定所分离的菌株MCA01属于恶臭假单胞菌。在以甲醛为唯一碳源的基础培养基中对菌株MCA01的甲醛耐受性及降解特性进行了研究,结果显示菌株MCA01至少能耐受1 600 mg·L~(-1)的甲醛浓度。当甲醛初始浓度≤1 000 mg·L~(-1)时,菌株MCA01能够在12 h内完全降解溶液中的甲醛;当甲醛浓度增至1 600 mg·L~(-1)时,菌株MCA01在24 h内对甲醛降解率为38.5%。菌株MCA01对甲醛具有良好的降解效率,该研究结果对殡仪场所内防腐废水等受甲醛污染环境的微生物治理具有较好的科学意义和应用价值。     

高效菲降解菌降解特性与蛋白组学研究

研究了各种条件下高效菲降解菌ZXl6对菲的降解特性,并借助一维与二维蛋白质组学分析技术,比对了菲诱导前后ZXl6蛋白表达差异.结果表明,该菌株利用菲生长的最适温度为35℃,最适pH为7.5,可以耐受并降解2 500 mgL-1菲.不能利用邻苯二甲酸,但可利用水杨酸和邻苯二酚.菌株细胞蛋白SDS-PAGE结果显示,菌株在菲诱导12 h后,出现两条M这l9x103和27x103的新增蛋白条带,从二维电泳图谱的相应分子量位置上发现3个新增表达蛋白,质谱分析与牛物信息学检索结果表明,2个点分别为芳香族加氧酶小亚基,1个点为顺式氯苯二氢二醇脱氢酶.     

高氯酸盐降解菌的分离鉴定及特性研究

研究降解高氯酸盐环境污染物微生物的形态特征、分子系统特征、生长特性和降解活性。利用PCA选择性培养基富集分离高氯酸根降解菌,观察其形态,进行生理生化测定,用分子克隆技术获得菌株的16SrDNA基因并测序,并对菌株的16S rDNA基因序列进行比对和系统发育分析,并用培养技术和离子色谱法检测菌株对底物的利用和降解情况。从镇江江滨和镇江新区污水处理厂的活性污泥中分别分离到一株能降解高氯酸根的菌株JD14和JD125。此菌株在24～30℃条件下较快地降解底物高氯酸盐,其中在24℃下,15d内可以将初始质量浓度为1600mg·kg-1的高氯酸根降解（86±6.5）%。对两菌株的16S rDNA基因进行了克隆和测序,并进行系统发育分析,结果表明菌株JD14和Dechloromonas sp.SIUL相似度高达100%,JD125和Dechlorospirillum相似度达97%,最后鉴定JD14和JD125属于高氯酸盐降解菌。此前国内并无降解高氯酸根微生物的报道,对今后在环境污染防治中开发利用此类细菌具有指导意义。     

一株高浓度苯胺、苯酚降解菌的分离鉴定及降解特性

从某生活污水厂活性污泥中分离到一株能够以苯胺或苯酚为唯一碳源、能源生长的高效降解菌菌株ANP.经形态特征、生理生化及16S rDNA序列分析,将该菌株鉴定为Delftia sp.进一步研究表明,该菌株利用苯胺生长的最适温度和pH分别为30℃和6.0,最适降解浓度为2000mgL-1;利用苯酚生长的最适温度和pH分别为35℃和8.0,最适降解浓度为1500mgL-1.苯胺、苯酚混合培养时该菌株对苯酚的降解过程要滞后于对苯胺的降解过程,但经过42h均能彻底降解.研究了ANP降解苯胺和苯酚的开环途径,苯胺芳环通过间位途径裂解,苯酚芳环则是通过邻位途径裂解.图4表1参18     

Biodegradation of carbofuran in sequencing batch reactor augmented with immobilised Burkholderia cepacia PCL3 on corncob

The performance of sequencing batch reactors (SBRs) augmented with immobilised Burkholderia cepacia PCL3 on corncob for biodegradation of carbofuran in basal salt medium (BSM) was studied. A 2.0-L SBR with a working volume of 1.5 L was operated for a total cycle of 48 h, consisting of 1.0 h fill phase, 46 h react phase and 1.0 h decant phase. The initial pH of the feed medium was 7.0. Air was fed into the reactor at a controlled flow rate of 600 mL·min ^?1. The effect of hydraulic retention time (HRT) (14 to 6 days) on carbofuran-degradation efficiency was investigated at a carbofuran concentration in the feed medium of 20 mg·L ^?1. The shortest HRT resulting in complete degradation of carbofuran was 8 days. At 75% of the optimum HRT (6 days), the effects of biostimulation using organic amendments, i.e. molasses, cassava pulp, rice bran and spent yeast, and the effect of carbofuran concentration in the feed medium (20–80 mg·L ^?1) were investigated. The optimum conditions for SBRs were an initial carbofuran concentration of 40 mg·L ^?1 and 0.1 g·L ^?1 of rice bran as a biostimulated amendment. Complete degradation of carbofuran with a first-order kinetic constant (k ₁) of 0.044 h^?1 was achieved under these optimum conditions.     

1株贫营养好氧反硝化菌的分离鉴定及其脱氮特性

从水库底泥样品中,以硝酸盐为唯一氮源进行驯化、分离筛选出1株能在贫营养及好氧条件下进行高效反硝化的菌株PY8,经过电镜形态学观察、生理生化和16S rDNA序列分析,并基于16SrDNA序列结果,构建了该菌株的系统发育树,最终确定菌株PY8为根瘤菌Rhizobiumsp.。考察了初始pH值、温度、C/N、初始硝酸钠质量浓度、投菌量对菌株PY8硝酸盐还原活性的影响,以及该菌株的异养硝化性能。结果表明,在pH6.0～10.0,温度25～30℃,C/N1.0～9.0,初始硝酸钠质量浓度0.01～0.50g·L-1,投菌量1%～15%时,菌株PY8培养72h后的硝氮去除率可达到95%以上。另外,该菌株具有同时硝化-反硝化作用,在培养过程中氨氮去除率可达到58%左右。实验结果表明,菌株PY8在微污染水体生物脱氮领域中具有很大的应用潜力。     

含油废水中一株高效油脂降解菌的筛选和鉴定

从中国海洋大学餐厅下水道废水中分离、筛选获得9株具有油脂降解能力的菌株。菌株ss-11油脂降解能力最强,在初始筛选条件下其降解率达47.29%。对ss-11的降解条件进行初步研究,结果表明,在初始豆油量为1.0mL·L-1,初始pH为7.5,摇床转速为140r·min-1,37℃下培养72h,该菌的油脂降解率达87.55%。通过菌落形态、生理生化特征、16SrDNA序列系统发育分析,将其鉴定为产酸克雷伯氏菌（Klebsiell aoxytoca）。     

Promotive effect of pyridine on indole degradation by activated sludge under anoxic conditions

Batch experiments were carried out to investigate the promotive effect of pyridine on indole degradation under denitrifying conditions. The seed sludge was obtained from a local coal-coking wastewater treatment facility and was acclimated in the laboratory. Indole and pyridine were supplemented to the synthetic wastewater at different ratios. The optimum ratio of chemical oxygen demand (COD) to nitrate (C/N) was 8.4–8.9 for both denitrification and indole and pyridine degradation. At a temperature of 28°C and pH of 7.0–7.5, the nitrate reductase activity (NRA) was in the best state. The addition of pyridine could promote NRA and the degradation of indole. When the initial concentration of indole was 150 mg/L, the concentration ratio of indole to pyridine was in the range of 1–10. Under optimum C/N conditions, the degradation of indole could be described with pseudo-zero-order kinetics. There was no accumulation of nitrite during the reaction. When the concentration ratio of pyridine to indole was less than 0.25 with an increase in the pyridine proportion, there were more significant augment rates for NRA and the degradation of indole than the situation when the concentration ratio was more than 0.25.     

红球菌Chr-9降解吡啶和苯酚的特性

研究了红球菌(Rhodococcus)Chr-9菌株在基础盐培养基中降解吡啶和苯酚的特性,分析了菌株降解苯酚和吡啶间的差异.结果表明,菌株Chr-9能够在72 h内将基础盐培养基中的吡啶(200 mg L-1)和苯酚(200 mg L-1)完全降解,同时利用吡啶和苯酚进行生长.菌株降解吡啶的最适温度为35℃,最适pH为8.0.菌株降解吡啶和苯酚的速度与底物的初始浓度呈负相关;在无其它氮源的基础盐培养基中,菌株能够利用吡啶和苯酚协同生长.图7参12     

Biodegradation of paclobutrazol by a microbial consortium isolated from industrially contaminated sediment

Biodegradability of the plant growth retardant paclobutrazol by a microbial consortium in which Pseudomonas was the predominant strain was investigated in batch culture. The consortium which had been isolated from an industrially contaminated sediment was proven to be useful for the treatment of effluents containing paclobutrazol. Paclobutrazol was degraded by the pure isolated strain of Pseudomonas sp. as well as the microbial consortium. Paclobutrazol was utilized as the sole source of carbon and energy. Sixty percent of the paclobutrazol was degraded by the microbial consortium from an initial concentration of 54 mg L^?1 within 48 h and more than 98% of an initial concentration of 3.4 mg L^?1 was degraded within 36 h. The optimum temperature and pH were determined to be 30°C and 7.0, respectively. A pure strain of a bacterium, isolated from the enrichment culture was identified as Pseudomonas sp. The microbial consortium was tolerant of high pH and could degrade paclobutrazol faster than the pure strain. The degradation rate of this plant growth regulator in an aerobic environment was greater than that under anaerobic conditions.     

一株能产生聚乙烯醇降解酶的委内瑞拉链霉菌

以聚乙烯醇(PVA)为唯一碳源,从土壤中筛选到1株能产生胞外PVA降解酶的放线菌GY1.根据扩增出的该菌株的16SrDNA全序列在GenBank中的比较结果,结合生理生化实验、细胞化学成分及菌落形态分析,确定该菌为委内瑞拉链霉菌(Stretopmycesvenezuelae).GY1菌株以PVA为唯一碳源时,产生的PVA降解酶活性达到120UL-1,是以葡萄糖或可溶性淀粉为唯一碳源时的10倍.当在PVA培养基中添加1gL-1至3gL-1的葡萄糖时,该菌株细胞量和PVA降解酶总酶活随着葡萄糖添加量的增加而增加,最大细胞量是未添加葡萄糖时的2.4倍,最高总酶活是未添加葡萄糖时的2.6倍,而单位质量细胞产生PVA降解酶的能力提高不大.但当添加的葡萄糖浓度从3gL-1增至10gL-1时,总酶活随着细胞量的增加出现下降趋势,同时单位质量细胞产生PVA降解酶的能力也大大降低.在相同PVA聚合度下,高醇解度PVA比低醇解度PVA更有利于GY1菌株的生长及产生PVA降解酶.图5表1参16     

Assessment of phenol-degrading ability of Acinetobacter sp. B9 for its application in bioremediation of phenol-contaminated industrial effluents

Successful bioremediation of a phenol-contaminated environment requires application of those microbial strains that have acquired phenol tolerance and phenol-degrading abilities. A newly isolated strain B9 of Acinetobacter sp. was adapted to a high phenol concentration by growing sequentially from low- to high-strength phenol. The acclimatised strain was able to grow and completely degrade up to 14?mM of phenol in 136?h. The degradation rates were found to increase with an increase in the phenol concentration from 2.0 to 7.5?mM. The strain preferred neutral to alkaline pH range for growth and phenol degradation, with the optimum being pH 8.0. The optimum temperature for phenol degradation was found to be in the range of 30–35°C. Transmission electron micrographs showed a disorganised and convoluted cell membrane in the case of phenol-stressed cells, showing a major effect of phenol on the membrane. Enzymatic and gas chromatography-mass spectrometry studies show the presence of an ortho-cleavage pathway for phenol degradation. Efficient phenol degradation was observed even in the presence of pyridine and heavy metals as co-toxicants showing the potential of strain in bioremediation of industrial wastes. Application of strain B9 to real tannery wastewater showed 100% removal of initial 0.5?mM phenol within 48?h of treatment.     

Methylobacterium sp. YAL-2对乙酰甲胺磷的降解特性

从有机磷生产厂家的下水道污泥中分离出一株对高浓度和低浓度乙酰甲胺磷都具有高效降解能力的寡营养菌YAL-2,根据形态、生理生化和16S rRNA基因系统发育分析,将菌株YAL-2鉴定为Methylobacterium sp.降解特性实验表明,菌株YAL-2能利用乙酰甲胺磷为唯一碳源生长和降解;在添加了甲醇的无机盐培养基中,84 h可完全降解300mg L-1乙酰甲胺磷,24 h将50 mg L-1和10 mg L-1乙酰甲胺磷降至非检测水平;4 d能完全去除100 mg L-1甲胺磷,5 d分别降解58.4%和40.6%的100 mg L-1乐果、敌敌畏.小青菜农药残留去除实验显示,菌株YAL-2可在7 d内将乙酰甲胺磷和甲胺磷将至限量水平.结果表明,将菌株YAL-2应用于保证果蔬等食品的食用安全是可行的.