萘降解菌MQ合成靛蓝的研究

利用萘降解菌(丛毛单胞菌)MQ完整细胞对吲哚进行转化合成靛蓝,结果表明菌株MQ具有较好的靛蓝合成能力,靛蓝产量在4h左右即达到稳定.薄层色谱和高效液相色谱分析结果表明,合成的蓝色产物为靛蓝.考察了反应温度及摇床转速对靛蓝合成的影响,结果表明最适温度和转速分别为25℃和150r·min-1.随后,采用表面响应法确定了菌株MQ合成靛蓝的最优条件:菌株接种量OD6602.16,吲哚浓度200.55mg·L-1,pH=6.91.在最优条件下,菌株MQ合成靛蓝的产量为53.05mg·L-1,比初始条件下的产量提高了179%.     

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.     

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.     

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.     

Pseudomonas sp.fz胞外分离物降解四溴双酚A

四溴双酚A(TBBPA)是全球消耗量最大的溴系阻燃剂。通过活性物质的定位、分离和TBBPA产物的分析等步骤,对Pseudomonas sp.fz胞外分离物降解TBBPA进行了研究。结果表明,存在于胞外的活性物质通过异丙基断裂和脱溴两条途径降解TBBPA。通过超滤,Sephadex G-10分离纯化得到了具有降解活性的小分子物质,分子量约为376~456 Da,初步鉴定为短肽类物质。这种小分子物质具有很好的热稳定性(30~80℃),在弱酸性条件下(2.0~5.0)活性较高,其活性还受金属离子、氧气和抑制剂(Na N3)的影响。此外,部分纯化的小分子物质在一定条件下能够产生羟基自由基(·OH)。     

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     

附生假单胞菌存在下不同光照时间对铜绿微囊藻生长与磷代谢的影响

通过设置8/16、12/12、16/8不同光暗比,分析了附生细菌存在下不同光照时间对铜绿微囊藻(M icrocystis aeruginosa)生长及其与附生假单胞菌(Pseudom onassp.)磷代谢之间关系的影响。结果表明:光照时间越长,铜绿微囊藻生长越快,16 h光照下的比增长速率为8 h光照下的1.6倍。铜绿微囊藻的快速生长促进了附生细菌中磷的释放;藻细胞增殖越快,附生细菌释放的磷越多。铜绿微囊藻对数生长期末,8、12、16 h光照下附生细菌磷含量分别降至对数生长初期的87.8%、78.6%和64.9%。铜绿微囊藻对附生细菌磷释放的促进作用是由藻细胞生长对磷的消耗再吸收导致的。