联苯代谢对微生物的生长胁迫及分裂抑制

以联苯/多氯联苯降解菌株R04（Rhodococcus sp.R04）和几种模式微生物为研究对象，利用高效液相色谱、荧光显微镜、扫描电子显微镜等分析微生物在联苯及其代谢物培养条件下细胞分裂和形态的变化.结果表明联苯及其代谢产物对红球菌R04和几种模式微生物细胞的分裂有抑制作用，并对部分微生物形态有影响.与前体-联苯及其代谢产物2-羟基-6-酮基-6-苯基-2，4-己二烯酸相比，2，3-二羟基联苯对G⁺、G^-细菌，或是酵母细胞分裂都有较强的抑制和形态的改变.2，3-二羟基联苯导致R.R04和缺陷型R.R04细胞形成不完整隔膜的比例增加；造成96.4%的大肠杆菌BL21细胞表面凹陷，胞质内容物流失，菌体体积缩小；导致枯草芽孢杆菌89.6%的细胞体积明显缩小；导致金黄色葡萄球菌基本没有细胞能形成完整的分裂隔膜；导致红酵母细胞能进行出芽生殖的比例从64.2%降低到19.3%，但对其细胞形态无明显改变.联苯代谢物2，3-二羟基联苯对红球菌R04及其它微生物细胞分裂和增殖的抑制作用比其前体-联苯强，建议在研究环境化合物与微生物互作时，应考虑环境化合物代谢的毒性效应.     

Effect of commercial pesticides on plant growth-promoting activities of Burkholderia sp. strain L2 isolated from rhizosphere of Lycopersicon esculentum cultivated in agricultural soil

The in vitro study was carried out to examine the toxicity of four commercially used pesticides (phorate, mancozeb, chlorpyrifos, and endosulfan) at the recommended dose (1X) and higher field doses (1.5X, 2X, 2.5X, and 3X) on plant growth-promoting activities of Burkholderia sp. strain L2, isolated from pesticide-treated rhizosphere of Lycopersicon esculentum. At the recommended dose, level of toxicity of all the pesticides to phosphate solubilization activity and indole-3-acetic acid (IAA) production was less detrimental as compared to exceeding doses. At maximum dose rate of each pesticide, the percentage decrease in phosphate solubilization and IAA production over control was found in order of chlorpyrifos < mancozeb < endosulfan < phorate. Present study proves that Burkholderia sp. strain L2 could be used as a potential microbial inoculant in the agricultural soil having persistent pesticide residues.     

Growth and Foliar Nitrogen Status of Four Plant Species Exposed to Atmospheric Ammonia

A chamber study was conducted to evaluate the growth response and leaf nitrogen (N) status of four plant species exposed to continuous ammonia (NH₃) for 12 weeks (wk). This was intended to evaluate appropriate plant species that could be used to trap discharged NH₃ from the exhaust fans in poultry feeding operations before moving off-site. Two hundred and forty bare-root plants of four species (Juniperus virginiana (red cedar), Gleditsia triacanthos var. inermis (thornless honey locust), Populus sp. (hybrid poplar), and Phalaris arundinacea (reed canary grass) were transplanted into 4- or 8-L polyethylene pots and grown in four environmentally controlled chambers. Plants placed in two of the four chambers received continuous exposure to anhydrous NH₃ at 4 to 5 ppm while plants in another two chambers received no NH₃. In each of the four chambers, 2 to 4 plants per species received no fertilizer while the rest of the plants were fertilized with a 100 ppm solution containing 21% N, 7% phosphorus, and 7% potassium. The results showed that honey locust was the fastest-growing species. The superior growth of honey locust among all species was also supported by its total biomass, root, and root dry matter (DM) weights. For all species there was a trend for plants exposed to NH₃ to have greater leaf DM than their non-exposed counterparts at 6 (43.0 vs. 30.8%; P = 0.09) and 12 wk (47.9 vs. 36.6%; P = 0.07), and significantly greater (P ≤ 0.05) leaf N content at 6 (6.44 vs. 3.67%) and 12 wk (7.05 vs. 3.51%) when exposed to NH₃. Numerically greater leaf DM due to NH₃ exposure was also consistently measured in poplar at both sampling periods. Hybrid poplar, as well as honey locust and reed canary grass, deposited 1.5 to 2-fold greater N in their leaves than red cedar tissues as a result of NH₃ exposure compared to non-exposed plants. Regardless of the effect of NH₃ on foliar color and damage score of the plants, the increase of foliar N content (g 100 g^?1 of fresh foliage weight) after NH₃ exposure at 6 and 12 wk was 0.45 and 0.87 for grass,1.25 and 1.34 for locust, and 2.67 and 6.09 for poplar. However, only honey locust likely benefited from ambient NH₃ as indicated by its consistent leaf color quality and lower damage score, compared with other species that were adversely affected by atmospheric NH₃.     

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%.     

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.     

理化因素对锰细菌Arthrobacter chlorophenolicus MN1409氧化Mn2+的影响

Arthrobacter chlorophenolicus MN1409是1株分离自锰矿样品的高效锰氧化细菌。为了建立该菌株氧化Mn2+的适宜条件,研究了接种量、装液量、摇床转速、温度、pH值、Fe2+初始质量浓度和Mn2+初始质量浓度等理化因素对其锰氧化效率的影响。结果表明,接种量、装液量、转速和Mn2+初始质量浓度在一定范围内变化对锰氧化率影响不大;而温度、pH值和Fe2+初始质量浓度变化对锰氧化率有较大影响。当接种量为4%,装液量为60 mL,温度为25℃,摇床转速为100r/min,pH值为7,且有一定的Fe2+存在时,Arthrobacter chlorophenolicusMN1409对锰的氧化效率最高。     

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     

芽孢杆菌发酵产碱性果胶酶温度控制策略

采用自行筛选获得一株产碱性果胶酶芽孢杆菌WSH03-09,在小型发酵罐中研究了不同温度对碱性果胶酶分批发酵的影响,结果表明,在恒定39℃条件下,可获得最高酶活5．39u／mL,各温度条件下的菌体干重相差不多,最终均能达11．5g／L左右;在发酵前期,控制温度41℃时最有利于菌体的生长,而在产物合成期,控制37℃有利于获得较高的产物合成比速,在此基础上,提出分阶段温度控制策略,采用此温度控制策略进行碱性果胶酶的发酵,碱性果胶酶酶活达5．99u／mL,比采用单一温度下的最大值提高了11％,其它各项指标也有较大提高．图6表1参6