微囊藻毒素降解菌的分子鉴定和特性研究

在成功筛选出1株能够降解微囊藻毒素(Microcystins,MCs)菌株的基础上,利用16S rDNA的PCR扩增和序列分析对其进行了分子鉴定。由BLAST序列比对及进化树分析可将该菌株定属至鞘氨醇单胞菌USTB-01(Sphingopyxissp.USTB-01),这是我国首次筛选出高效降解MCs的鞘氨醇单胞菌。在鞘氨醇单胞菌USTB-01降解MC-RR和MC-LR活性研究方面发现,温度30℃和pH7.0条件下该菌降解MCs的速率最大,日均降解MC-RR和MC-LR分别达到了23.4mg/L和7.9mg/L,在进一步去除水体中的MCs的基础和应用研究方面都具有非常重要的意义。     

鞘氨醇单胞菌完整细胞对溴氨酸的好氧降解

文章考察了鞘氨醇单胞菌QYY完整细胞对溴氨酸的好氧降解条件,并对其降解途径进行了分析。研究表明,该完整细胞在最佳条件为:温度30℃、初始pH7.0～8.0、摇床转速150r/min,磷酸缓冲溶液浓度为0.01～0.04mol/L时将溴氨酸完全脱色后产生3种产物:2-氨基-3-羟基-5-溴苯磺酸、2-氨基-4-羟基-5-溴苯磺酸为终产物;邻苯二甲酸为中间产物。其脱色过程中,大部分溴氨酸直接降解为终产物,仅有一小部分溴氨酸在降解过程中形成邻苯二甲酸。邻苯二甲酸则通过苯甲酸代谢途径开环。     

滇池沉积物菌群对微囊藻毒素的厌氧生物降解

好氧微生物降解已经被证明是微囊藻毒素(MC)自然转化的主要途径,但是厌氧降解的作用尚不明确.为了揭示这一降解过程,研究了滇池沉积物中混合菌群在厌氧条件下对MCLR的降解能力,并考察了环境因素和外加营养源对该过程的影响.结果表明,厌氧条件下MCLR在2 d内从5 mg/L迅速降解到检测限以下,说明该菌群在厌氧条件下对MCLR具有较强的降解能力,并且可以利用MCLR作为唯一氮源.在实验温度范围内,MCLR的降解速率随着温度的升高而增大.酸性条件下MCLR的厌氧降解缓慢(pH=5.0)甚至停止(pH=3.0),而中性(pH=7.0)和碱性(pH为9.0、11.0)条件下降解速率没有显著差异.单独添加葡萄糖可以产生酸性物质而使体系的pH下降,从而抑制MCLR的降解,但是同时添加硝酸盐可以消除这一影响.单独添加硝酸盐对MCLR的厌氧降解也有显著的抑制作用,说明硝酸根在这一过程中未被MCLR厌氧降解菌用作最终电子受体.以上结果表明,厌氧降解可能是沉积物中MCLR转化的另一重要途径,该过程在MCLR污染治理方面具有潜在的应用价值.     

滇池水环境中微囊藻毒素的生物降解

利用从滇池蓝藻水华生物量中提取的微囊藻毒素试验溶液,接种滇池沉积物的微生物,研究其在有氧条件下的生物降解过程.结果表明,水体中的微囊藻毒素易被生物降解,其降解反应服从方程C=A/(1+Be-^Ct).当温度在12～25℃,加入的沉积物量为1~10g时,藻毒素粗提液的平均降解反应速率为3.181.13d^-1,平均半衰期t^1/2为2.661.27d,且藻毒素的生物降解速度随反应温度和沉积物量的增加而提高.研究结果还表明,生物降解是去除滇池水环境中微囊藻毒素的一个重要机制.     

筛选菌种酶催化降解微囊藻毒素的特点

从云南滇池底泥中筛选出1株能够高效降解微囊藻毒素(Microcystins,MCs)的纯菌种.该菌无细胞提取液能进行降解MC-RR和MC-LR的酶促反应,并有最终产物的积累.在pH 6～8范围内酶对MC-RR和MC-LR的催化降解活性较高,Cu²⁺、Mn²⁺和Zn²⁺对降解MCs的酶促反应没有明显的促进作用.     

藻毒素生物降解技术的研究进展

藻毒素是湖泊水华现象滋生的有害有毒污染物,它严重恶化了饮用水水源地的水质。有效控制水华爆发及去除藻毒素是环境科学领域的一个难题。近些年来,利用生物降解技术去除藻毒素已经取得了显著的效果。笔者对国内外利用生物降解技术去除微囊藻毒素的最新研究进展进行了综述,并提出了控制藻毒素的新途径。     

一株降解微囊藻毒素的缺陷短波单胞菌的分离鉴定及降解特性研究

从太湖水华腐烂蓝藻中筛选出一株能够降解MC-LR的细菌,将其编号为Q3.经形态、生理生化及16S rRNA基因序列分析鉴定为缺陷短波单胞菌(Brevundimonas diminuta).研究发现,在实验条件下该菌能以MC-LR为唯一碳源和氮源生长;7 d内可将初始浓度为0.96 mg·L-1的MC-LR降解为0.37 mg·L-1,降解效率达到61.5%.同时,本研究首次发现缺陷短波单胞菌能够降解藻毒素,并且在此菌种中扩增出了降解过程的关键基因mlr A,推测该菌可能与已报道的Sphingomonas sp.ACM-3962等菌具有相同的降解机制.     

降解微囊藻毒素菌种的筛选和活性研究

研究了滇池底泥和表层水体中的微生物菌群降解微囊藻毒素(Microcystins, MCs)的能力差异,发现底泥中的微生物菌群对MCs有更强的生物降解能力.采用从滇池水华蓝藻细胞中提取提纯的微囊藻毒素作为微生物生长的唯一碳源和氮源,先后经过液体和固体培养基培养后,通过挑取单克隆菌落,分别从底泥中筛选出了能够降解MC-RR和LR的5种不同微生物菌种.其中筛选的菌种D降解MCs的能力最强,在3 d内可将初始浓度分别为60.1 mg·L^-1和38.7 mg·L^-1的MC-RR和LR全部降解,日均降解MC-RR和LR的速率分别高达20.0 mg·L^-1和12.9 mg·L^-1.     

利用流式细胞术研究鞘氨醇单胞菌GY2B降解菲过程中细菌表面特性的变化

微生物与污染物接触是生物降解的第一步,为进一步了解降解过程中有机污染物对降解菌的影响,通过对不同条件下鞘氨醇单胞菌GY2B(Sphingomonas sp.GY2B)降解菲的研究,并结合Propidium Iodide(PI)染料和流式细胞仪分析不同底物及污染物浓度对GY2B菌体细胞表面性质的影响.结果表明,随着菲的降解,PI染色的GY2B菌株细胞增多,说明细菌膜结构受到一定的破坏,通透性不断增强.污染物浓度越高,降解菌的膜完整性的破坏越为严重.60 h时,300 mg·L-1浓度条件下,染色细胞/未被染色细胞的比值可达12.44,而在100 mg·L-1和1.2 mg·L-1浓度条件下,比值分别为1.95和1.11.同时降解过程中细菌细胞的傅里叶红外光谱检测,死亡、受损和完整细胞的区分以及Zeta电位分析也进一步验证了细菌细胞表面性质的改变.利用流式细胞术与染料结合分析降解过程中细菌细胞膜完整性的变化,从单细胞水平上了解细菌降解污染物过程中其细胞表面性质的改变,有助于更好地研究GY2B对菲的降解机制.     

溴氨酸降解菌株的分离鉴定及特性研究

从污泥样品中分离得到一株溴氨酸降解菌,该菌株能够以溴氨酸为唯一碳、氮源及能源生长.通过形态、生理生化特性分析以及对16SrDNA序列进行同源比较,鉴定该菌株属于鞘氨醇单胞菌属,定名为Sphingomonasxenophaga .菌株的最适降解与生长条件为:pH =7 0 ,温度3 0℃,转速15 0r·min- 1 ,接种量8% .在最佳条件下,溴氨酸的降解率可达90 %以上,菌株可耐受的溴氨酸浓度为10 0 0mg·L- 1 .通过分析菌株对水杨酸、邻苯二酚及邻苯二甲酸的利用情况,推测该菌株可能通过邻苯二酚的代谢途径降解溴氨酸.     

Microbial biodegradation of microcystin-RR by bacterium Sphingopyxis sp. USTB-05

A strain,USTB-05,isolated from Lake Dianchi,China,degraded the cyanobacterial toxin microcystin-RR(MC-RR) at the rate of 16.7 mg/L per day.Analysis of 16S rDNA sequence showed that the strain was Sphingopyxis sp.Enzymatic degradation pathways for MC-RR by Sphingopyxis sp.USTB-05 were identified.Adda-Arg peptide bond of MC-RR was cleaved and then a hydrogen and a hydroxyl were combined onto the NH 2 group of Adda and the carboxyl group of arginine to form a linear molecule as intermediate product within the ...     

Cloning and expression of the first gene for biodegrading microcystin LR by Sphingopyxis sp. USTB-05

Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins (MCs) produced by harmful cyanobacteria. Based on the isolation of a promising bacterial strain of Sphingopyxis sp. USTB-05 for biodegrading MCs, we for the first time cloned and expressed a gene USTB-05-A (HM245411) that is responsible for the first step in the biodegradation of microcystin LR (MC-LR) in E. coli DH5αup, with a cloning vector of pGEM-T easy and an expression vector of pGEX-4T-1, respectively. The cell-free extracts (CE) of recombinant E. coli DH5αup containing USTB-05-A had high activity for biodegrading MC-LR. The initial MC-LR concentration of 40 mg/L was completely biodegraded within 1 hr in the presence of CE with a protein concentration of 0.35 mg/mL. Based on an analysis of the liquid chromatogram-mass spectrum (LC-MS), the enzyme encoded by gene USTB-05-A was found to be active in cleaving the target peptide bond between 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-deca-4,6-dienoic acid (Adda) and arginine of MC-LR, and converting cyclic MC-LR to linear MC-LR as a first product that is much less toxic than parent MC-LR, which offered direct evidence for the first step on the pathway of MC-LR biodegradation by Sphingopyxis sp. USTB-05.     

微生物降解藻毒素的研究进展

介绍了近几年国内外非生物法处理和控制藻毒素污染的研究近况,阐述了微生物降解藻毒素的可行性、菌种筛选、降解机理及参与细菌降解藻毒素的酶,提出了今后在微生物控制及处理藻毒素方面值得进一步研究和需要关注的问题。     

Burkholderia sp.FDS-1降解硝基苯酚类化合物的特性研究

从长期受农药污染的土壤中分离到1株高效杀螟硫磷降解菌株Burkholderia sp.FDS^-1,它能以3-甲基-4-硝基苯酚(MNP)为唯一碳源和氮源进行生长.该菌能在9h内将0.6mmol.L-1MNP完全转化成邻甲基对苯二酚(MHQ),并能继续降解MHQ.该菌降解MNP的最适温度和pH分别为30℃和7.0,菌株降解MNP的速率和起始接种量呈正相关.降解MNP的酶为诱导酶.     

细菌降解非离子表面活性剂的研究

从石油污染的土壤中分离到２株能利用非离子表面活性剂ＡＥＯ－９和ＳＡ－２０作为唯一碳源生长的细菌。这２株菌被定名为假单胞杆菌５２（Ｐｓｅｕｄｏｍｏｎｏｓ５２）和Ｗｅｅｋｓｅｌｌａ６．最适于这２株菌利用非离子表活剂ＡＥＯ－９的培养条件是乙酸胺作为氮源,ｐＨ７．３０℃添加少量葡萄糖促进表活性的降解,降解速率的研究表明,当ＡＥＯ－９的初始浓度为５０００ｍｇ／Ｌ时,５２号菌和６号菌能在２周内将其去除８５％,     

菌株scl-2对苯线磷、哒嗪硫磷和丙溴磷降解的研究

研究了有机磷农药降解菌株Arthrobacter sp.scl-2对苯线磷、哒嗪硫磷和丙溴磷的降解特性,结果表明菌株scl-2可以利用苯线磷、哒嗪硫磷和丙溴磷为唯一磷源生长.菌株scl-2降解苯线磷、哒嗪硫磷和丙溴磷的最适温度均为30℃,哒嗪硫磷的降解在初始pH8时速度最快,而丙溴磷和苯线磷的最适初始pH均为7,偏酸偏碱均不利于菌株对苯线磷、哒嗪硫磷、丙溴磷的降解.利用气相色谱-质谱联机鉴定了苯线磷、哒嗪硫磷、丙溴磷的降解产物,分别为3-甲基-4-甲硫基苯酚、6-羟基-2-苯基-2-氢哒嗪酮和4-溴-2-氯苯酚.     

Biodegradation of benzo[a]pyrene in soil by Mucor sp. SF06 and Bacillus sp. SB02 co-immobilized on vermiculite

Two indigenous microorganisms, Bacillus sp. SB02 and Mucor sp. SF06, capable of degrading polycyclic aromatic hydrocarbons （PAHs） were co-immobilized on vermiculite by physical adsorption and used to degrade benzo[a] pyrene （BaP）. The characteristics of BaP degradation by both free and co-immobilized microorganism were then investigated and compared. The removal rate using the immobilized bacterial-fungal mixed consortium was higher than that of the freely mobile mixed consortium. 95.3% of BaP was degraded using the co-immobilized system within 42 d, which was remarkably higher than the removal rate of that by the free strains. The optimal amount of inoculated co-immobilized system for BaP degradation was 2%. The immobilized bacterial-fungal mixed consortium also showed better water stability than the free strains. Kinetics of BaP biodegradation by co-immobilized SF06 and SB02 were also studied. The results demonstrated that BaP degradation could be well described by a zero-order reaction rate equation when the initial BaP concentration was in the range of 10--200 mg/kg. The scanning electronic microscope （SEM） analysis showed that the co-immobilized microstructure was suitable for the growth of SF06 and SB02. The mass transmission process of co-immobilized system in soil is discussed. The results demonstrate the potential for employing the bacterial-fungal mixed consortium, co-immobilized on vermiculite, for in situ bioremediation of BaP.     

大豆磷脂对不动杆菌降解硝基苯的影响

研究了液相体系中不同用量的大豆磷脂对不动杆菌降解硝基苯的影响.大豆磷脂可以作为不动杆菌的唯一碳源,在一定程度上支持其生长.实验表明,在适宜不动杆菌生长的400 mg/L硝基苯质量浓度下,加入500 mg/L大豆磷脂对不动杆菌生物量和硝基苯的完全降解时间无影响,而加入250,1 000 mg/L大豆磷脂却延缓了细菌的生长和硝基苯的降解,这主要是由于它与硝基苯形成竞争代谢或是由于硝基苯在胶束中的分配降低了其生物可利用性;当硝基苯质量浓度约为600 mg/L时,加入大豆磷脂(高于临界胶束浓度)显著促进了细菌的生长和硝基苯的降解,并且促进效果与加入量成正比,这主要是因为胶束的分配作用对硝基苯产生了脱毒效果,另外大豆磷脂作为辅助碳源增加了生物量,也促进了对硝基苯的代谢分解.      

Biodegradation of geosmin in drinking water by novel bacteria isolated from biologically active carbon

Three strains of Gram-negative bacteria capable of removing geosmin from drinking water were isolated from biologically active carbon and identified to be Chryseobacterium sp., Sinorhizobium sp. and Stenotrophomonas sp. based on physio-biochemistry analysis and 16S rRNA gene sequence analysis. Removal e ciencies of 2 mg/L geosmin in mineral salts medium were 84.0%, 80.2% and 74.4% for Chryseobacterium sp., Sinorhizobium sp. and Stenotrophomonas sp., respectively, while removal e ciencies of 560 ng/L geosmin in filter influent were 84.8%, 82.3% and 82.5%, respectively. The biodegradation of geosmin was determined to be a pseudo first-order reaction, with rate constants at 2 mg/L and 560 ng/L being 0.097 and 0.086 day??1, 0.089 and 0.084 day??1, 0.074 and 0.098 day??1 for the above mentioned degraders, respectively. The biomass of culture in the presence of geosmin was much higher than that in the absence of geosmin.