1株铜绿假单胞菌对芘的降解特性及代谢途径

多环芳烃（PAHs）因其具有"三致"作用对生态系统产生潜在威胁.微生物降解是多环芳烃降解的主要途径之一,筛选出能高效降解多环芳烃的菌株是微生物修复技术的关键.本文采用富集培养的方法从多环芳烃污染的污泥中分离到1株以芘为唯一碳源的菌株LX2,经形态观察、生理生化和16S rDNA鉴定,LX2属于铜绿假单胞菌（Pseudomonas sp.LX2）.菌株在含芘浓度为50 mg·L^-1的无机盐液体培养基中培养21 d对芘的降解效率达32.1%.经GC-MS分析发现,Pseudomonas sp.LX2降解芘的中间代谢产物主要有4,5-二氢芘、2''-羟基苯丙酮、苯酚、原儿茶酚.基于鉴定的代谢产物得出芘通过"萘"和"邻苯二甲酸"两种不同的途径被铜绿假单胞菌（Pseudomonas sp.LX2）降解.     

嗜盐高效降酚菌株Halomonas sp.H17的筛选及降解苯酚特性

从巴丹吉林沙漠盐湖沉积物中分离获得1株在高盐环境下高效降解苯酚菌H17.分析了H17生理生化特性、16S rDNA基因序列、苯酚降解特性及动力学,结果表明,H17属于盐单胞菌属(Halomonas sp.),能在0～20%的盐度下有效降解苯酚,每升外加适量的碳源(葡萄糖浓度0.8 g)和复合氮源(KNO_3 1 g、NH_4Cl 5 g、酵母提取物0.2 g和胰蛋白胨0.2 g)能够促进H17的生长及降解苯酚能力.在温度为30℃、pH 7～8、盐度5～10%的条件下,H17均能高效降解苯酚,最高降解率可达到88.5%.该菌株降解苯酚动力学符合Halane模型,经拟合其生长参数为μ_(max)=0.31 h~(-1),K_S=191.63 mg·L~(-1),K_i=683.05 mg·L~(-1).研究显示H17具有在高盐环境下降解和耐受苯酚的能力,同时对环境有较强的适应性,体现出其在高盐含酚废水实际处理中具有良好的应用价值.     

聚苯乙烯微塑料对铜绿假单胞菌生物膜形成和结构变化的影响

微塑料在水环境中的污染情况日益严重,对水生生物的生长发育造成严重影响,而目前有关微塑料对生物膜形成影响机制的研究十分有限.为探究聚苯乙烯微塑料（PS-MPs）对铜绿假单胞菌生物膜形成发育的影响,选用不同浓度和粒径PS-MPs进行暴露试验,研究其对生物膜内生物量、氧化应激水平、生物膜结构和群体感应系统的影响.结果表明,PS-MPs诱导严重氧化应激反应并抑制生物膜形成和发育,粒径越小,产生的抑制作用越强烈,抑制效果表现为0.1 μm>0.5 μm≈1 μm>5 μm.PS-MPs通过与细胞接触造成严重物理损伤,形成的生物膜厚度显著减小且结构稳定性遭到破坏,膜内细菌通过分泌胞外聚合物来抵御PS-MPs的胁迫作用.PS-MPs进一步通过干扰铜绿假单胞菌群体感应系统,下调关键基因lasI、lasR、rhlI和rhlR的表达水平,抑制信号分子和相关毒力因子的合成与分泌,降低细菌对毒性作用的防御功能,最终影响生物膜形成和结构稳定性.     

Purification and Properties of a Poly (β-hydroxybutyrate) Depolymerase From Penicillium sp.

An extracellular poly (β-hydroxybutyrate) (PHB) depolymerase was purified from a Penicillium sp. DS9701-09a by centrifugation, ultrafiltration, precipitation and gel filtration chromatography. The specific activity of the purified enzyme was 37.9-folds higher than that of the culture supernatant and the recovery yield was 11.8%. The PHB deploymerase molecular mass was 44.8 kDa from analysis of both Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Matrix-assisted laser desorption-time-of-flight (MALDI-TOF) mass spectrometer. The isoelectric point of 6.7 for the enzyme was determined by a two-dimensional electrophoresis. The optimum enzyme activity was observed at a temperature of 50 °C and pH 5.0. The apparent K _m of the enzyme was found to be 1.35 mg/mL. The PHB depolymerase consisted of 16 kinds of normal amino acids. The secondary structure of the enzyme was determined by CD spectrum. α-helix and β-turn were found to be 66% and 34% for the enzyme without ammonium sulphite. Chemical inhibition on the PHB depolymerase activity was examined and EDTA was found to have a significantly inhibitory effect.     

Promotion of hexadecyltrimethyleamine bromide to the damage of Alexandrium sp. LC3 by cupric glutamate

The effect of hexadecyltrimethyleamine bromide （HDTMAB） on the removal of A lexandrium sp. LC3 under cupric glutamate stress was investigated. Toxic effect of cupric glutamate on A lexandrium sp. LC3 was significantly promoted in the presence of HDTMAB, especially at 3.0 cmc of HDTMAB. It was found that the sulfhydryl group content of the cell decreased, while the malonaldehyde content and membrane permeability increased when A lexandrium sp. LC3 was treated with HDTMAB and cupric glutamate complex, compared with cupric glutamate alone. The data suggest that HDTMAB might stimulate the damage of A lexandrium sp. LC3 by enhancing the membrane permeability.     

苯并噻吩脱硫菌株的筛选及脱硫活性研究

从孤岛油田油浸土样中筛选到1株能降解苯并噻吩(BT)的脱硫菌,经初步鉴定该菌为戈登氏菌属(Gordona sp.).实验证明:该菌能以类似于4S途径脱除BT及其衍生物中的硫,但是不能脱除二苯并噻吩(DBT)及其衍生物中的硫.GC-MS分析表明该途径的终产物为邻羟基苯乙醛或其异构体苯并呋喃.在以BT为唯一硫源的培养基中30℃培养48h,Gordona sp.C-6能降解0.15mmol/L的BT,终产物占发酵培养基中BT加入量的50%,其余BT在有氧培养过程中挥发.通过Matlab拟合曲线确定以邻羟基苯乙酸为标准品进行产物定量检测的方法.     

重组荧光假单胞菌BioP8在环境中消长动态与安全性评价

通过选择分离培养和PCR鉴定,对转苏云金芽孢杆菌(Bacillus thuringiensis,Bt)杀虫蛋白基因荧光假单胞菌BioP8及其天然受体菌P303在温室自然土壤和田间大白菜根际、叶面的环境生存竞争能力进行研究.结果表明,在温室自然土壤中,可培养细菌总量在108 CFU/g土(湿重)左右,40 d内P303和BioP8群落总量分别能维持在105 CFU/g土(湿重)和104 CFU/g土(湿重)左右,100 d后检测不到残留; 在大白菜根际,可培养细菌总量在1010 CFU/g根(湿重)左右,30 d内P303和BioP8可维持在106 CFU/g根(湿重)和105 CFU/g根(湿重),75 d后检测不到残留; 在大白菜叶面,可培养细菌总量在103～106 CFU/ cm2叶片之间波动,处理后3 d的供试菌菌量迅速由105 CFU/ cm2叶片降至最低(103 CFU/cm2叶片和102 CFU/cm2叶片左右),之后回升并以(104和103)CFU/ cm2叶片维持一段时间(第5-15 d),30 d检测不到残留.P303及其工程菌株在温室自然土壤、田间大白菜根际和叶面的定殖时间至少分别为60 d、50 d和15 d,它们的消长动态相似,在温室自然土壤、田间大白菜根际的定殖能力明显强于叶面,BioP8定殖能力弱于出发菌P303.     

Effects of cultivation strategies on the cultivation of Chlorella sp. HQ in photoreactors

Heterotrophic cultivation caused high level of ROS and high lipids accumulation. HMTC is the best culture strategy for improving the microalgal biomass. Chlorella sp. HQ had great nutrient removal capacity under five culture strategies. The effects of cultivation strategies (including autotrophic cultivation (AC), heterotrophic cultivation (HC), fed-batch cultivation (FC), heterotrophic+ autotrophic two-stage cultivation (HATC), and heterotrophic+ mixotrophic two-stage cultivation (HMTC)) on the growth and lipid accumulation of Chlorella sp. HQ and its total nitrogen (TN) and total phosphorus (TP) removal in secondary effluent were investigated in column photoreactors. The results showed that the TN and TP removal rates ranged between 93.72%–95.82% and 92.73%–100%, respectively, under the five different strategies. The microalgal growth potential evaluated by the maximal growth rate (R_max) was in the order of HMTC>HC>FC>AC>HATC. The values of biomass, total lipid yield, triacylglycerols (TAGs) yield, and total lipid content of the microalga cultivated in the last 5 d increased significantly, but the TAGs productivities of the five strategies were lower than those in the first 7 d. Compared with all the other cultivation strategies, the TAGs productivity and yield after 12 d of cultivation under the heterotrophic condition reached the highest values accompanying the highest level of intracellular reactive oxygen species (ROS), in which the TAGs yield reached 40.81 mg/L at the end of the cultivation period. The peaks in TAGs yield and ROS level suggested that HC was beneficial for lipids accumulation via regulating the cellular redox status and exerting ROS stress on microalgal cells. In summary, HMTC was the best cultivation strategy for improving the microalgal biomass and HC was the best strategy for microalgal TAGs accumulation to produce biodiesel.     

Ralstonia sp.strain U2菌株对萘的趋化作用

一株降解萘的细菌Ralstonia sp. strain U2对萘具有趋化现象,萘、水杨酸都可以作为其对萘产生趋化现象的诱导物.U2菌对水杨酸、龙胆酸同样都具有趋化性,但这种趋化性是组成型的,不需要诱导.图3参22     

海洋苯酚降解菌Candida sp.P5的分离鉴定及其降解特性

从海洋沉积物中分离、筛选到一株能以苯酚作为唯一碳源和能源的酵母菌P5.根据菌落特征、菌体形态、生理生化特性和18S rDNA序列分析,确定菌株P5为假丝酵母菌属(Candida sp.).该菌株最适宜生长和降解苯酚的条件为:温度25℃,pH6.0～7.0,摇床转速100r/min,需氧;菌株P5能在较高浓度的苯酚条件下生长,在72h内可以降解95%以上的苯酚.对苯酚代谢途径和相关酶的研究发现,菌株P5主要在邻苯二酚1,2-双加氧酶作用下通过邻位途径进行苯酚代谢.图7表2参24