外源甜菜碱提高恶臭假单胞菌(Pseudomonas putida)DLL-1耐盐性的研究

研究了外源甜菜碱对恶臭假单胞菌(Pseudomonas putida)DLL-1耐盐性的影响并对其渗透保护机制进行了初步的探讨;结果表明培养基中添加甜菜碱可以改善DLL-1细胞在高盐培养基中的生长情况,添加150mg/L的甜菜碱可以使DLL-1在1.2mol/L NaCl的基础盐培养基中生长,添加10mg/L的甜菜碱就足以显著缩短渗透胁迫条件下DLL-1细胞的延滞期和代时,增加生长量;和不添加对照相比,延滞期由24h缩短到6h,代时由60min缩短到35.7min,最大生长量OD610由1.29增长到1.57。在渗透胁迫条件下,细胞从外界快速吸收外源甜菜碱来代替自身相容性溶质的合成。

趋化信号转导基因cheA突变对Pseudomonas putida DLL-1甲基对硫磷的趋化性及原位降解的影响

Pseudomonas putida DLL-1是一株甲基对硫磷（MP）高效降解菌株,同时对MP具有趋化性.cheA基因是菌株趋化信号转导过程中负责编码组氨酸激酶的基因,为了研究菌株趋化性在农药原位降解中的作用,通过基因打靶的方式使P.putida DLL-1染色体上单拷贝的cheA基因失活,成功地获得了MP的趋化突变株P.putida DAK,突变株与野生菌株生长能力没有显著差异.通过土壤盆钵试验（MP浓度为50mg/kg）,发现在灭菌与未灭菌土壤中趋化突变株对MP的降解能力低于原始出发菌株DLL-1约20%～30%,说明菌株DLL-1趋化性的丧失会减慢其对农药的降解,趋化性在农药的原位降解过程中发挥重要作用.

不同生长期的Pseudomonas putida 5-x细胞对工业废水中Cu^(2+)的吸附

研究了不同生长阶段的Cu2 + 吸附细菌Pseudomonasputida 5 x对Cu2 + 的吸附容量 .结果表明在低硫培养基中 ,当P putida 5 x细胞生长刚进入对数生长期时 ,单位细胞对Cu2 + 的吸附溶量最低 .而当生长进入稳定生长期的后期或死亡期的早期时 ,单位细胞对Cu2 + 的吸附量最大 .实验结果还表明 ,高浓度的Cu2 + 对对数期细胞的吸附能力有一定的抑制 ,而对死亡期的细胞影响较小 .早期研究已证明细胞对重金属离子的吸附和细胞表面某些对重金属离子具有亲和性的功能基团 (如某些膜蛋白及羰基化合物 )有关 .利用透视电子显微镜观察了不同生长阶段的细胞表面结构及对铜离子的吸附效果 ,发现生长后期细胞表面吸附有较多的Cu2 + 。

固定化Pseudomonas putida CGMCC3830转化3-氰基吡啶制备烟酸

采用固定化Pseudomonas putida CGMCC3830转化3-氰基吡啶制备烟酸。选择海藻酸钠作为包埋材料进行固定化,考察固定化细胞的制备条件、转化条件以及批次稳定性。结果表明,优化的固定化条件为:海藻酸钠2 g/dL,氯化钙0.4 g/dL,固化时间6 h;最适转化条件为:温度35℃,pH 7.0,底物浓度100 mmol/L。批次转化实验结果显示,当底物浓度为100 mmol/L时,固定化细胞重复使用10次,酶活仍保留59.1%,产物烟酸的得率为91.8 g/g细胞干重,而游离细胞的使用3次后,酶活下降至45.6%。

Pseudomonas putida KT2442制备中长链聚羟基烷酸酯

初步探讨了利用PseudomonasputidaKT2442制备中长链聚羟基烷酸酯(MCL PHAs)的工艺,考察了不同碳源对菌体生长和产物合成的影响。在2.5L发酵罐上采用高密度细胞培养制备:以油酸为碳源,细胞干重达120g L,MCL PHAs质量浓度达40g L,产物质量分数为34%,生产速率为0.965g·L-1·h-1;以玉米油水解物为碳源,细胞干重达105g L,MCL PHAs达28g L,产物质量分数为26.5%,生产速率为0.667g·L-1·h-1。以玉米油水解物为碳源所得细胞浓度、产物浓度与以油酸为碳源所得结果比较接近,有利于降低生产成本。

Pseudomonas putida MnB1氧化分解菱锰矿的实验研究

自然界中,菱锰矿氧化形成锰的氧化物矿物是非常普遍的现象,在菱锰矿被氧化分解发生物相转变的过程中,碳酸盐溶解和锰的氧化往往同时发生,微生物可能起着催化作用。选取锰氧化模式菌株Pseudomonas putida Mn B1和广西梧州菱锰矿,通过菱锰矿在该细菌作用下发生转变的实验,利用场发射扫描电镜、扫描透射X射线显微成像等分析方法,研究了矿石表面形貌变化以及锰元素在细胞上的分布特征。结果表明细菌显著促进的菱锰矿的溶解,在此基础上,进一步探讨了细菌在菱锰矿氧化过程中的贡献,本实验结果丰富了次生锰矿床的微生物成因研究。

Pseudomonas putida ZWL73经龙胆酸途径分解代谢3-羟基苯甲酸

一株4-氯硝基苯(4-CNB)的完全降解菌Pseudomonas putidaZWL73能利用3-羟基苯甲酸(3-HBA)为惟一碳源和能源生长,而且ZWL73在对3-HBA降解的过程中,有中间产物龙胆酸生成;酶学分析的分光光度法证实,ZWL73通过GSH依赖型的龙胆酸途径代谢3-HBA,并且该途径需经3-HBA或龙胆酸诱导.

Pseudomonas putida S1海藻糖合成酶表达质粒的构建及诱导条件优化

采用PCR方法从Pseudomonas putidaS1中克隆出编码海藻糖合成酶的基因treS,并与质粒pQE30T相连,构建了表达质粒pQE-TS2。将此重组质粒转化宿主菌E.coliM15进行诱导表达。十二烷基磺酸钠-聚丙烯酰胺凝胶SDS-PAGE电泳结果表明,treS基因在大肠杆菌中获得了高效表达。通过对诱导温度、诱导剂浓度、加诱导剂时间和诱导时间的优化研究,在菌液生长至OD600值为0.6时,加入诱导剂IPTG至终浓度0.01 mmol/L,20℃诱导20 h,蛋白的表达量达到每克干细胞89 mg的蛋白,粗酶液酶活达到19 U/mL。

Degradation of o-chloronitrobenzene as the sole carbon and nitrogen sources by Pseudomonas putida OCNB-1

A bacterial strain that utilized o-chloronitrobenzene （o-CNB） as the sole carbon, nitrogen and energy sources was isolated from an activated sludge collected from an industrial waste treatment plant. It was identified as Pseudomonas putida based on its morphology, physiological, and biochemical characteristics with an automatic biometrical system and the 16S rRNA sequence analysis. Microcosm study showed that the biodegradation of o-CNB was optimized at culture medium pH 8.0 and 32℃. At these conditions, the strain degraded 85% of o-CNB at a starting concentration of 1.1 mmol/L in 42 h. o-Chloroaniline was identified as the major metabolite with high performance liquid chromatography （HPLC） and gas chromatography-mass spectrometry （GC-MS）. The study showed that o-CNB degradation by Pseudomonas putida OCNB-1 was initiated by aniline dioxyenase, nitrobenzene reductase and catechol-l,2- dioxygenase.

Pseudomonas putida S1海藻糖合成酶基因在大肠杆菌中的克隆表达

利用PCR和TA克隆方法扩增和克隆得到了恶臭假单胞菌Pseudomonas putidaS1的海藻糖合成酶基因treS。对其进行序列分析表明,其编码区含有2067bp,编码含688个氨基酸残基的蛋白质,其核苷酸序列和蛋白质序列与来源于其它假单胞菌属细菌的海藻糖合成酶的序列表现出了较高同源性。将该基因序列与表达载体pQE30T连接,构建重组质粒pQE30T-TS,并将其转化至E.coliM15菌株中。重组菌株经诱导表达后SDS-聚丙烯酰胺凝胶电泳结果显示有明显的分子量约77.5kD的特异蛋白条带出现。经测定酶活力达19U/mL,约是原始菌株P.putidaS1的50倍。

优化细胞表面组分与结构以提高P.putida 5-x细胞的重金属吸附能力

研究了生物吸附剂Pseudomonasputida 5 x细胞的最佳制备条件 ,以提高其对重金属的吸附能力。并用Pseudomonasputida 5 x细胞为生物吸附剂处理含铜离子城市污水。实验结果表明 ,在优化的条件下 ,Pseudomonasputida 5 x细胞对铜离子的吸附容量可达 87.3mg g ,经生物吸附过程处理后 ,废水中Cu2 + 的含量降低至微量级 ,而且该吸附剂可有效循环使用

Zinc Oxide Nanoparticles in Bacterial Growth Medium: Optimized Dispersion and Growth Inhibition of <i>Pseudomonas putida</i>

The majority of nanoparticles tend to agglomerate in bacterial growth media. Thus, nanoparticle-specific characteristics can get lost. To investigate the influence of nanoparticles on bacteria, these particles should remain in their nanoparticulate state. The present study demonstrates the stabilization of commercially available zinc oxide (ZnO) with sodiumhexametaphosphate (SHMP) in bacterial growth medium (LB) to avoid agglomeration of these particles after the addition to LB. This established method is appropriate to stabilize ZnO agglomerates as small as 43 nm. The method of fractionated centrifugation was used to obtain stable agglomerates (also stable in the presence of bacteria) with different mean diameters. The SHMP-stabilized ZnO inhibits the growth of Pseudomonas putida with increasing concentration (up to 500 mg/L) and decreasing agglomerate size (43 - 450 nm).

Mass production, fermentation, formulation of <i>Pseudomonas putida</i>for controlling of die back and phomopsis diseases on grapevine

The export oriented agricultural and horticultural crops depends on the export of residue free produce and has created a great potential and demand for the incorporation of biopesticides in crop protection. PGPR with wide scope for commercialization includes Pseudomonas putida. Pseudomonas putida was effective in reducing die back caused by a fungus Botryodiplodia theobromae and phomopsis or arm death, caused by a fungus, Phomopsis viticola of grapevine in vitro and in vivo. Pseudomonas putida showed optimum Siderophore pseudobactin production at 72 h, and growth peak at 120 h. glycerol as a source of carbon and yeast as a source of nitrogen Pseudomonas putida was formulated using polysaccharide dextran starch, which led to the elongation of conservation and the effectiveness of the biofungicide. Pseudomonas putida was very effective as biocontrol agents to reduce the die back and phomopsis diseases on grapevine. The obtained results showed that both bacterial treatments significantly increased the growth parameters as well as fruits dry weights and yield of grapevine.

Polyhydroxyalkanoate Production by <i>Pseudomonas putida</i>KT217 on a Condensed Corn Solubles Based Medium Fed with Glycerol Water or Sunflower Soapstock

Pseudomonas putida KT217 was grown on a complex medium comprised of co-products of the ethanol and biodiesel industries to assess the organism's capability to produce medium-chain-length polyhydroxyalkanoate (mcl-PHA). The growth phase was carried out in a medium containing 400 g/L condensed corn solubles (CCS), supplemented with ammonium hydroxide as a nitrogen source. Following the exponential phase, co-products of the biodiesel industry (soapstock and glycerin) were fed into the reactor to trigger PHA production. When glycerin was added to the bioreactor (75 g/L total addition), the final cell dry weight (CDW) and PHA content were 30 g/L and 31%, respectively. The monomeric composition in the PHA formed was relatively uniform throughout incubation with 3-hydroxydecanoate dominating. When a total of 153 g/L of sunflower soapstock was added to the bioreactor in a fed-batch manner, the final CDW and PHA content were 17 g/L and 17%, respectively. Following addition of soapstock the monomeric composition of the polymer changed dramatically, with the 3-hydroxyoctanoate monomer becoming dominant and greater unsaturation present in the PHA.

Isolation and molecular characterization of a novel <i>pseudomonas putida</i>strain capable of degrading organophosphate and aromatic compounds

A bacterial strain designated in this study as POXN01 was found to be capable of degrading the synthetic organophosphorus pesticides paraoxon and methyl parathion. The strain was initially isolated through enrichment technique from rice field soil near Harlingen, Texas. Phylogenetic analysis based on 16S rRNA, gyrB and rpoD gene alignments identified the POXN01 isolate as a new strain of Pseudomonas putida, which is closely related to the recently discovered nicotine-degrading strain Pseudomonas putida S16. While being unable to metabolize nicotine, the POXN01 isolate was observed to actively proliferate using monocyclic aromatic hydrocarbons, in particular toluene, as nutrients. Search for the genetic determinants of paraoxon catabolism revealed the presence of organophosphorus-degrading gene, opd, identical to the one from Sphingobium fuliginis (former Flavobacterium sp. ATCC 27551). Assimilation of aromatic compounds likely relies on phc ARKLMNOPQ gene cluster for phenol, benzene and toluene catabolism, and on benRABCDKGEF cluster for benzoate catabolism. The observed versatility of POXN01 strain in degradation of xenobiotics makes it useful for the multi-purpose bioremediation of contaminated sites in both agricultural and industrial environmental settings.

Strain Pseudomonas putida PAO-1 Isolate with Polyphosphate Accumulating and Elongation Ability

Filamentous bacteria(FB)overgrowth is an important cause of sludge bulking in wastewater treatment plants(WWTPs).However,to date,methods for the cultivation and preservation of isolated FB in the laboratory have not been completely described.Furthermore,research on whether FB can function as phosphorus accumulating organisms(PAOs)is limited.In this study,a pure strain,a Pseudomonas putida PAO-1(P.putida PAO-1)isolate with phosphorus removal functions was isolated from the biofilm of an alternating anaerobic/aerobic biofilter(AABF),and its physiological characteristics were studied.Nitrate or nitrite could be used by the strain P.putida PAO-1 as electron acceptors for denitrification during phosphorus anoxic uptake,and 0.63 mg NO-3-N was consumed to reduce 1 mg soluble orthophosphate(SOP)by P.putida PAO-1.The strain P.putida PAO-1 consumed phosphorus within the optimal pH range of 6 to 8 and the temperature range of 25℃to 35℃.Cell deformity was a main morphological trait of the strain P.putida PAO-1,and it could elongate(with an elongation rate of 300%-500%)when it was subjected to oligotrophic or high-salt stress(15 g·L-1 NaCl).The findings in this study provide a microbiological reference for understanding the special characteristics of a denitrifying PAO.

Statistical optimization of process variables by response surface methodology to enhance phenol degradation by Pseudomonas putida (NCIM 2102)

Removal efficiency of phenol from aqueous solutions was determined using Pseudomonas putida (NCIM 2102). Experiments were made as a function of pH (4 - 9), temperature (28 - 36oC), and agitation speed (100 - 200 rpm). Optimization of these three process parameters for phenol degradation was studied. Statistically designed experiments using response surface methodology was used to get more information about the significant effects and the interactions between these three parameters. A 23 full-factorial central composite design was employed for experimental design and for analysis of the results. A second order polynomial regression model, has been developed using the experimental data. It was found that the degrading potential of P.putida (NCIM 2102) was strongly affected by the variations in pH, temperature and agitation speed. The experimental values were in good agreement with the predicted values and the correlation coefficient was found to be 0.9871. The optimum process conditions for maximizing phenol degradation were recognized as follows: pH (7.49), temperature (29.99oC), and agitation speed (138.89) rpm.

Pseudomonas putida细胞对微囊藻毒素-LR的胁迫响应

将1.0g/L的微囊藻毒素-LR(MC-LR)降解菌恶臭假单胞菌(Pseudomonas putida)置于含不同浓度MC-LR的体系中,研究了体系中菌体细胞完整性和生物量的变化,考察了MC-LR对细胞的氧化胁迫以及抗氧化酶的响应.结果表明,MC-LR能够增大P.putida细胞质膜通透性,造成膜损伤,导致胞内物质外流,使细胞完整性遭到破坏;同时,MC-LR能够引起P.putida细胞的氧化胁迫,随着毒素暴露时间的延长,活性氧自由基(ROS)和膜脂过氧化产物丙二醛(MDA)含量显著升高,具有明显的剂量效应.超氧化物歧化酶(SOD)活性在MC-LR的诱导下有一个先升后降的过程,表现为对低浓度污染物的主动响应,而高浓度(2.5mg/L)MC-LR作用5d后,ROS积累到相当高水平,对细胞代谢功能造成破坏,使SOD活性下降,并加速细胞的死亡,P.putida生物量与对照相比,下降了将近50%.

Enhanced Biological Phosphorus Removal with Pseudomonas putida GM6 from Activated Sludge

提高的生物的磷移动(EBPR ) 方法广泛地从废水为磷移动被采用，然而，很少对它的微生物、分子的机制被知道。因此，在一家大规模市政的污水处理工厂预言并且控制 EBPR 过程的恶化是困难的。使用的这研究在实验室孤立的新奇紧张， Pseudomonas putida GM6，它有高磷酸盐积累能力并且能从败坏的系统很快恢复并且在活性污泥提高磷移动的能力。与 gfp 基因标记的紧张 GM6，被称为 GMTR，被交付进定序低效率的批反应堆(SBR ) 的凳子规模，到调查 GMTR 的殖民和磷的移动。在 21 天以后，而磷移动率是 96% ，在污泥的全部的细菌的 GMTR 的比例到达了 9.2% ，与大约 0.2 mg L 的自河集中 ? 1。在有 GMTR 的增加的反应堆，磷快速被移开，在在厌氧的条件下面的 1 h，并且在在氧气的条件下面的 2 h。这些证据 EBPR 过程是特征的。现场测试与低磷移动能力在一台医院污水处理设备被进行。二十 -- 一天在 Pseudomonas putida GM6 被增加以后，自河磷集中在 0.3 mg L 附近留下了 ? 1，相应于 96.8% 的移动率。它因此被表明为废水的快开始和改进的那 Pseudomonas putida GM6couldbeused 生物的磷移动，它为潜在的大规模工程申请提供了一个科学基础。

StyS激酶自磷酸化对Pseudomonas putida B3中靛蓝生物合成的调节作用

在恶臭假单胞菌Pseudomonas putida B3中,靛蓝生物合成关键酶基因styAB上游存在一个二元调控系统StyS-StyR。StyS蛋白属于激酶家族,是磷酸化信号转导的重要媒介,但激酶StyS的自磷酸化传导机制对靛蓝生物合成的调节作用尚未探明,因此,研究以Pseudomonas putida B3中激酶蛋白StyS作为研究对象,发现了两个磷酸化结合位点,构建了磷酸化位点突变株。通过分析野生菌株P.putida B3、styS基因缺失菌株P.putida B3-ΔstyS、styS基因回补菌株P.putida B3-ΔstyS-8和磷酸化位点突变株P.putida B3-ΔstyS-1之间的靛蓝产量及酶活发现,P.putida B3产量为10.14 mg/L,P.putida B3-ΔstyS-8产量为3.63 mg/L,磷酸化位点突变菌株无激酶活性且失去了产生靛蓝的能力。结果表明,StyS自磷酸化作用在靛蓝发酵体系中起重要作用。研究结果将为靛蓝生物合成途径的进一步研究提供参考。