α-蒎烯降解菌株的筛选及降解特性

从生产萜烯树脂的废水中分离到一株能降解α-蒎烯的菌株,通过16S r RNA序列鉴定和构建系统发育树分析,该菌株为海洋小单胞菌(Micromonospora marine),命名为HKY。研究了温度、p H值、α-蒎烯的浓度对这株菌降解α-蒎烯的影响,结果表明,菌株在25~40℃对α-蒎烯都有较好的降解,其中在30℃降解率最高,可达到90.21%。初始p H在6~8之间时降解率较高,其中p H为7时,降解率最高可达到93.43%。当α-蒎烯在100 mg/L以内时,培养48 h后没有残留,200 mg/L和300 mg/L时培养液中有残留,500 mg/L基本对该菌处于抑制状态。同时研究了该菌株的底物谱,结果表明该菌株能够降解一些分子结构较为简单或者与α-蒎烯结构相似的工业有机污染物。运用该菌处理生产萜烯树脂污水处理厂的出水,能够对COD有一定的去除作用。     

潘多拉菌LX-1菌株对二氯甲烷的降解特性研究

从某污水处理厂的活性污泥中分离筛选到一株能以二氯甲烷为唯一碳源和能源生长的好氧降解菌LX-1(Pandoraea pnomenusa LX-1).单因素试验表明,适宜菌株LX-1生长的pH和环境温度分别为7.0和35℃,培养基里添加1%的NaCl有助于LX-1的生长及对DCM的降解,二氯甲烷的最大耐受浓度达到了1500mg·L-1.菌株LX-1降解1mg二氯甲烷能产生0.8083mg Cl-和0.3838mg CO2,脱氯率和矿化率分别达到了96.8%和74.15%.培养液中可溶性有机碳含量随降解时间变化呈上升趋势,pH呈下降趋势,结合相关文献报道,推测菌株在代谢二氯甲烷的过程中产生了水溶性小分子酸类(如甲酸)等结构较为简单的有机物,它们最终将被矿化为CO2、H2O和细胞生物量.     

1株1,2-二氯乙烷降解菌的分离及降解特性研究

从浙江某化工厂污水处理池的活性污泥中筛选分离到1株能以1,2-二氯乙烷(1,2-dichloroethane,1,2-DCA)为唯一碳源和能源生长的菌株T-2,根据菌株的形态特征、生理生化特性及16S rRNA基因序列分析,该菌株被鉴定为Starkeya sp.研究表明,Starkeya sp.T-2生长和降解1,2-DCA较适宜的温度为30℃,较适宜的pH为7.0～8.0;菌株降解1,2-DCA的过程遵循Haldane动力学模型,其最大比生长速率μmax为0.065 h-1,最大比降解速率vmax为0.54 h-1;菌株降解1,2-DCA的耐受浓度为500 mg.L-1,细胞产率系数(以1,2-DCA计)为0.191 mg.mg-1,菌株T-2利用1,2-DCA生长的过程中能同时把1,2-DCA最终矿化为CO2和H2O,矿化率为45%.     

荧光假单胞菌代谢α-蒎烯过程中表面活性物质的定向积累及其性状分析

从处理α-蒎烯的生物滴滤塔中分离到一株荧光假单胞菌PT,该菌株在代谢α-蒎烯的过程中能产生具有表面活性的物质.通过优化培养基,使得菌株在降解过程中能最大程度地积累表面活性的代谢产物,培养液的表面张力下降至30 mN·m-1.采用红外光谱、薄层色谱等分析方法对分离得到的油状产物进行鉴定,并结合α-蒎烯的微生物代谢途径,确定该表面活性物质为紫苏酸,分子式为C10H14O2.这类物质对柴油和正己烷有较明显地乳化作用,乳化指数达到40％～60％,对多环芳烃(菲、萘、芘)也有明显地增溶效果,在其临界胶束浓度时水中溶解度分别提高了2.65％、6.07％和10.0％.以上结果表明,该表面活性物质可替代化学表面活性剂广泛用于环境污染的修复治理中.     

1株氯苯高效降解菌的分离鉴定及降解特性

分离筛选到1株能以氯苯为唯一碳源和能源的菌株LW26,根据菌株的形态、生理生化特征、16S rRNA序列分析以及Biolog鉴定,确定该菌株为戴尔福特菌(Delftia tsuruhatensis),其为新发现的具有氯苯降解能力的菌株,并且该菌株能降解BETX、正己烷和环己烷等常见的有机污染物.实验考察了温度、pH、氯苯初始浓度、Cl-浓度等因素对菌株生长和降解性能的影响.结果表明,该菌株较为适宜的生长和降解条件为:温度25℃、pH 7.0;底物耐受浓度高达500 mg·L~(-1);当Cl-浓度超过0.14 mol·L~(-1)时,菌株生长会受到抑制.利用Haldane模型对实验数据拟合得到菌株LW26的最大比生长速率μmax和最大比降解速率γmax分别为0.42 h~(-1)和2.53 h~(-1).利用GC-MS进行中间产物分析,结果表明菌株LW26降解氯苯的过程中产生邻氯苯酚,结合邻苯二酚双加氧酶活性分析,推测氯苯经历邻位开环、脱氯、氧化等过程,最终矿化为CO2或转化为生物质.     

生物过滤塔和生物滴滤塔净化α-蒎烯性能比较

分别采用以木屑/泥炭为填料的生物过滤塔(BF)和以聚氨酯小球为填料的生物滴滤塔(BTF)净化α-蒎烯废气,比较两者的挂膜时间及对α-蒎烯的降解性能.结果表明,采用气液相联合方法,过滤塔和滴滤塔分别在21d和27d内完成挂膜;扫描电镜观察表明,填料上生物膜菌群生长良好,优势菌为杆菌和球菌.在α-蒎烯进口浓度80～2200mg.m-3、空床停留时间(EBRT)29～102s条件下,两者对α-蒎烯均有较好的去除效果,过滤塔与滴滤塔的最大去除负荷分别为50g.m-.3h-1和43g.m-.3h-1;滤塔中CO2生成量与α-蒎烯降解量之间呈线性关系,通过线性拟合得出过滤塔与滴滤塔的α-蒎烯矿化率分别为74%与68%,滤塔中减少的α-蒎烯主要被微生物利用而去除.菌落数(CFU)分析表明,在挂膜阶段滤塔微生物数量增长明显,稳定运行阶段菌落数随着EBRT的延长而增加,在EBRT102s条件下单位反应器空间内过滤塔和滴滤塔菌落数分别为5.52×1014cfu.m-3和1.84×1014cfu.m-3.     

苯酚降解菌的分离及降解特性研究

从扬子乙烯集团废水处理系统曝气池中的活性污泥驯化分离得到一株能快速降解苯酚的菌株,初步鉴定其为假单胞菌属菌株。该菌株在5℃-35℃范围内时都可以有效降解并矿化200mg／L的苯酚,最适宜的生长温度为25℃左右;菌株在pH为5～9范围内可以降解200mg／L的苯酚,偏碱性的条件下比酸性条件更适合细菌生长;培养过程中振荡速率大于120r／m时降解速率最大。当苯酚的初始浓度超过1000mg／L时,降解菌的生长受到抑制,不能有效降解苯酚。     

Pseudomonas sp. ZW代谢α-蒎烯的关键产物及其代谢酶鉴定

利用气相色谱-质谱分析检测了假单胞杆菌Pseudomonas veronii ZW在降解α-蒎烯过程中产生的10种主要中间代谢产物,完善了其代谢途径。通过紫外诱变的方法,成功获得一株α-蒎烯降解失活的突变株ZW-U,该菌株对某些中间产物丧失了降解能力。利用双向电泳技术从这2株菌中分离获得16种差异蛋白,并通过MALDI-TOF-MS进行分析,结果表明其中14种蛋白与细胞功能有关,另外2种蛋白(PKHD型羟化酶和芳基醇脱氢酶)在α-蒎烯的代谢过程中起重要作用,PKHD型羟化酶催化对伞花烃转化为5-甲基-2-异丙基苯酚和4-异丙基苯甲醇,4-异丙基苯甲醇在芳基醇脱氢酶的催化下转化为4-异丙基苯甲醛。这两种酶的发现,进一步验证了菌株ZW代谢α-蒎烯的途径,有利于今后对ZW降解α-蒎烯进行代谢调控。     

菌株Sphingomonas sp. FL降解溴氨酸的特性研究

分离了1株澳氨酸降解菌.其可以溴氨酸为唯一碳源进行降解并使其脱色,通过16S rRNA基因序列比较和生理生化特性分析,将其归为鞘氨醇单胞菌属.溴氨酸降解和菌株生长的最适条件为:温度30℃,pH 7.0,摇床转速100 r/rain,(NH4)2SO4作为氮源,在此条件下,溴氨酸(100 mg/L)在14 h内的脱色率可达99%.低浓度NaCl(<2%)对脱色有促进作用,而高浓度NaCl(≥2%)对脱色产生抑制.以Haldane底物抑制模型表征溴氨酸初始浓度对脱色的影响.确定当初始浓度为1 393.5 mg/L时可取得最佳比降解速率1.4 h-1.菌株不能将溴氨酸完全矿化,至反应终点52.4%的有机碳得到去除.利用GC-MS和HPLC-MS分析代谢产物显示.溴氨酸降解的中间产物是邻苯二甲酸,终产物可能为2-氨基-3-羟基-5-溴苯磺酸或2-氨基-4-羟基-5-溴苯磺酸,邻苯二甲酸可经3,4-二羟基苯甲酸途径进一步降解而被菌体利用.     

“真菌-细菌”复合菌剂的构建及其降解α-蒎烯、乙酸丁酯和邻二甲苯混合废气的性能研究

以α-蒎烯、乙酸丁酯和邻二甲苯3株高效降解菌为菌源(2株真菌和1株细菌),通过对比单菌单底物和多菌多底物降解效果,确定这3株菌之间不存在抑制效应,可用于构建"真菌-细菌"复合菌剂.考察了复合菌剂与活菌液的降解性能及对饥饿期的响应,结果表明复合菌剂和活菌液均能较完全地降解底物,但经历饥饿期后,复合菌剂对底物仍保持100%的降解率,而活菌液对α-蒎烯和邻二甲苯的去除率只有94.6%和62%.该菌剂具有较好的稳定性,常温保存180 d后对各初始浓度为120 mg·L-1的α-蒎烯、乙酸丁酯和邻二甲苯24 h内去除率为48.2%、95.1%和57.3%.将复合菌剂与活性污泥分别接种于生物过滤塔,复合菌剂能明显缩短生物膜形成时间,接种的反应器对底物总去除率维持在90%左右,而活性污泥只有60%.     

Removal characteristics and kinetic analysis of an aerobic vapor-phase bioreactor for hydrophobic alpha-pinene

Biofiltration is considered an effective method to control volatile organic compounds (VOCs) pollution. This study was conducted to evaluate the potential use of a bacterial biofilter packed with wood chips and peat for the removal of hydrophobic α-pinene. When inoculated with two pure degraders and adapted activated sludge, a removal efficiency (RE) of more than 95% was achieved after a start-up period of 11 days. The maximum elimination capacity (EC) of 50 g/(m³. hr) with RE of 94% was obtained at empty bed retention time (EBRT) of 102 sec. When higher α-pinene concentrations and shorter EBRTs were applied, the REs and ECs decreased significantly due to mass-transfer and biological reaction limitations. As deduced from the experimental results, approximately 74% of α-pinene were completely mineralized by the consortiums and the biomass yield was 0.60 g biomass/g α-pinene. Sequence analysis of the selected bands excised from denaturing gradient gel electrophoresis revealed that the inoculated pure cultures could be present during the whole operation, and others were closely related to bacteria being able to degrade hydrocarbons. The kinetic results demonstrated that the whole biofiltration for α-pinene was diffusion-limit controlled owing to its hydrophobic characteristics. These findings indicated that this bacterial biofiltration is a promising technology for the remediation of hydrophobic industrial waste gases containing α-pinene.     

Kinetic analysis and bacterium metabolization of α-pinene by a novel identified Pseudomonas sp. strain

Biodegradation has become a popular alternative remediation technology for its economic and ecological advantages. An aerobic bacterium (strain ZW) capable of degrading αup-pinene was isolated from a biofilter by a selective enrichment. Based on the 16S rRNA gene analysis and physiochemical properties, this strain was identified as Pseudomonas veronii. Under the optimized condition achieved by the response surface methodology (RSM), as well as pH 6.82, temperature 26.3\textcelsius and NaCl concentration 1.36%, almost 100% αup-pinene could be removed within 45 hr. Enzymatic biodegradation by the crude intracellular enzyme could be described well by the Michaelis-Menten model in which the maximum degradation rate V_max and the half-saturation constant K_m were calculated to be 0.431 mmol/(L. min) and 0.169 mmol/L, respectively. Activity assay of catechol suggested that the strain ZW possessed a catechol-1,2-dioxygenase and could decompose benzene-ring through ortho ring cleavage. Based on the identified intermediates by GC/MS, a new metabolic pathway was proposed, in which the final metabolites were some simpler organic and inorganic compounds. The present work demonstrated that the strain ZW would have a great application prospect for the remediation of αup-pinene-contaminated environment.     

一株氯氰菊酯降解真菌的筛选鉴定及其降解特性研究

从雅安市售砖茶中分离筛选出一株对氯氰菊酯具有较高降解能力的真菌菌株YAT,根据其形态学及ITS序列分析,将该菌鉴定为黑曲霉(Aspergillus niger).菌株YAT在PD培养基中168 h内对50 mg·L-1氯氰菊酯的降解率为54.83％,对氯氰菊酯的降解率与菌体生物量呈正相关.动力学研究表明,菌株YAT降解氯氰菊酯的过程符合一级动力学方程,在测试的底物(氯氰菊酯)浓度、温度、pH范围内,氯氰菊酯半衰期为85.750～281.958 h,较高温度及偏碱性条件有利于氯氰菊酯的降解,底物浓度越高,其半衰期越长.此外,菌株YAT对50 mg·L-1溴氰菊酯、氰戊菊酯、氟氯氰菊酯和氯菊酯的120 h降解率分别为27.53％、58.00％、53.23％和25.34％.     

一株苯乙烯高效降解菌的分离鉴定及降解特性研究

从某橡胶厂的活性污泥中筛选得到一株可高效降解苯乙烯的微生物菌株WJ,通过透射电镜(TEM)、生理生化试验、Biolog鉴定和16S rRNA基因的系统发育树建立等分析方法,鉴定该菌株为恶臭假单胞菌(Pseudomonas putida).通过环境单因素试验,确定该菌株WJ在温度30℃、pH值7.0的培养条件下具有良好的降解效果.通过不同初始浓度苯乙烯的降解及生长特性,确定该菌株的最大底物耐受浓度达到1500mg/L,且其生长动力学过程符合Haldane’s方程,相应动力学参数为: vmax=0.282h-1,Ks=23.57mg/L,Ki=1784.56mg/L.底物宽泛性试验表明,该菌株对苯系物(BTEX)、正己烯等与苯乙烯结构类似的碳氢化合物都具有良好的降解能力,表明其具备良好的底物宽泛性.该菌株对苯乙烯的氧化降解途径主要为乙烯基侧链的氧化.     

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.     

水华蓝藻生物质对Cu~(2+)的吸附动力学及吸附平衡研究

文章初步研究了水华蓝藻生物质对重金属离子Cu2+的吸附特性,包括生物吸附动力学和吸附等温线。生物吸附动力学结果表明,Cu2+的生物吸附可分为两个阶段,第一个阶段能较快地达到平衡,而之后的阶段进行缓慢。透析袋处理的蓝藻生物质在不同pH(3,4,5,6)的铜离子溶液条件下,吸附效率依次为62.4%,78.1%,71.2%,60.35%,吸附最大值为7.81 mg/g。水华蓝藻生物质对Cu2+的生物吸附平衡等温线可以用Freundlich模型精确拟合。水华蓝藻生物质在处理低浓度的含Cu2+废水方面有潜力。湖泊富营养化的优势在于产生低成本的、大量的、可收集的水华,这说明自然环境中的藻类资源如蓝藻水华生物质,可能作为商业中使用的生物吸附剂。     

Effect of illumination intensity and light application time on secondary organic aerosol formation from the photooxidation of α-pinene

Secondary organic aerosol (SOA) formation from hydroxyl radical (OH.) initiated photooxidation of α-pinene was investigated in a home-made smog chamber. The size distribution of SOA particles was measured using aerodynamic particle sizer spectrometer. The effects of illumination intensity and light application time on SOA formation for α-pinene were evaluated. Experimental results show that the concentration of SOA particles increased significantly with an increasing of illumination intensity, and the light...     

热脱附气相色谱质谱法测定空气中萜烯有机物

采用自动热脱附(ATD)和气相色谱质谱(GC/MS)联用技术,研究并建立了空气中11种萜烯有机物的测试方法。采用吸附剂Tenax TA进行吸附捕集时,α-蒎烯、莰烯、β-蒎烯极易发生穿透现象,三者的穿透率超过60%;与Tenax TA相比,吸附剂Carbopack B/CarbosieveTMS-Ⅲ可有效抑制萜烯有机物的穿透行为。ATD优化的操作参数为:一级脱附温度和时间分别选择为260℃和10 min,冷肼温度为-20℃,二级热脱附温度和时间分别为280℃和5 min。该方法测试结果准确,重现性和稳定性较好,方法的加标回收率为89%¹12%,相对标准偏差(RSD)小于11%。将该法用于室内空气中萜烯类有机物的测试,发现不同空气样品中均含有多种萜烯有机物,其总量约占总挥发性有机物(TVOC)的15%112%,相对标准偏差(RSD)小于11%。将该法用于室内空气中萜烯类有机物的测试,发现不同空气样品中均含有多种萜烯有机物,其总量约占总挥发性有机物(TVOC)的15%⁶5%,出现几率较大的有α-蒎烯、β-蒎烯、香叶烯、蒈烯、柠檬烯和柏木烯,松油醇较少出现。     

一株稻田土壤梭状芽孢杆菌的分离及其固氮产氢能力研究

为研究水稻土壤中协同固氮产氢菌的固氮产氢特性,拓展协同固氮产氢菌菌种资源,利用厌氧微生物富集培养、亨盖特厌氧滚管等技术,从华南稻田土壤中筛选到一株同时具有产氢和固氮能力的菌株BZ-1. 经形态学观察及16S rRNA基因序列系统发育分析,鉴定菌株BZ-1属于梭状芽孢杆菌属(Clostridium sp.). 通过测定产氢量、生物量、发酵产物及固氮酶酶活等,对菌株BZ-1的产氢能力及协同固氮产氢特性进行分析. 结果表明:菌株BZ-1发酵34.55 mmol/L葡萄糖可产生42.19 mmol/L H₂,主要副产物为丁酸(15.96 mmol/L)、乙酸(7.14 mmol/L)和乳酸(5.09 mmol/L);菌株BZ-1具有固氮酶活性,能够以N₂为唯一氮源进行生长;菌株BZ-1固氮产氢时,相比于添加7 mmol/L氯化铵的试验组,产氢量提高了14.71%,最大生物量降低了33.33%,乙酸产量提升了61.49%. 研究显示,菌株BZ-1在协同固氮产氢条件下固氮能力的提升、生物量的降低以及核心碳代谢途径的改变可能是其产氢量提升的原因. 固氮产氢菌株BZ-1的获得将为提高土壤肥力以及缓解铅、镉等重金属对农作物的胁迫作用提供新的思路.      

1株咔唑降解菌的分离、鉴定及其降解特性研究

利用微生物分离技术,从土壤中分离到1株咔唑降解菌KH-6.菌株KH-6能够以咔唑为唯一的碳源和能源良好地生长,同时还可以利用3-甲基咔唑、4-羟基咔唑和2,2′-二羟基联苯生长.经过对其形态特征、生理生化、以及16S rDNA序列分析,初步鉴定为黄杆菌属细菌.该菌株对咔唑的降解性是通过在液体培养基内菌体的增加及底物的减少来证实的,其利用咔唑生长的最适温度和pH分别为30℃和7.5.静止细胞反应试验表明,菌株KH-6除了咔唑以外还可降解其它杂环化合物.咔唑污染土壤中的生物降解实验表明,在灭菌土壤中90%的咔唑被降解,菌株KH-6对土壤中的咔唑有很好的生物降解效果.