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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

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微生物学报 2007年第3期47卷 471-476页

作者：文阳蒋建东邓海华蓝鸿李顺鹏南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室南京210095

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... 详细信息

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.

关键词：趋化性农药降解 Pseudomonas putida DLL-1 cheA基因甲基对硫磷

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

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微生物学报 2006年第1期46卷 154-157页

作者：何健蒋建东贾开志黄星李顺鹏南京农业大学生命科学学院微生物学系农业部农业环境微生物工程重点开放实验室南京210095

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... 详细信息

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。

关键词：外源甜菜碱渗透保护耐盐性 Pseudomonas putida DLL-1

海洋细菌Pseudomonas putida中具有抗硅藻附着的活性成分

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微生物学报 2013年第8期53卷 825-831页

作者：朱建生姜薇缪莉靳翠丽包卫洋扬州大学海洋科学与技术研究所扬州225127 中国科学院海洋生物资源可持续利用重点实验室广州510301

【目的】为发现天然的防污损物质,从分离自海绵Haliclona sp.的细菌Pseudomonas putida中寻找具有抗硅藻附着活性的化合物。【方法】综合菌落生长形态、扫描电镜及16S rDNA序列分析,鉴定细菌种属;采用活性(抗硅藻附着)-化学(薄层色谱、... 详细信息

【目的】为发现天然的防污损物质,从分离自海绵Haliclona sp.的细菌Pseudomonas putida中寻找具有抗硅藻附着活性的化合物。【方法】综合菌落生长形态、扫描电镜及16S rDNA序列分析,鉴定细菌种属;采用活性(抗硅藻附着)-化学(薄层色谱、二极管阵列高效液相和核磁共振氢谱)导向法对其中的活性组分进行分离纯化,波谱分析确定结构;采用抗硅藻附着活性模型对单体化合物进行活性复筛。【结果】该细菌鉴定为Pseudomonas putida;从其发酵液中分离得到6个环二肽,分别鉴定为环(亮氨酸-脯氨酸)(1)、环(亮氨酸-丙氨酸)(2)、环(苯丙氨酸-丙氨酸)(3)、环(缬氨酸-酪氨酸)(4)、环(丙氨酸-酪氨酸)(5)、环(丙氨酸-色氨酸)(6);化合物3和6在浓度为50μg/mL时具有明显的抑制硅藻附着活性(抑制率分别为50%和85%)。【结论】海洋细菌Pseudomonas putida中具有抗硅藻附着的活性化合物为环(苯丙氨酸-丙氨酸)和环(丙氨酸-色氨酸)。

关键词：抗硅藻附着 Pseudomonas putida 环二肽海洋细菌

以菌株Pseudomonas putida KT2440为宿主构建多功能农药降解菌

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应用与环境生物学报 2016年第6期22卷 1145-1149页

作者：易忠权王睿周潮洋陈雪婷洪青何健闫新李顺鹏南京农业大学生命科学学院南京210095 东南大学医学院附属盐城医院中心实验室盐城224000

通过富集培养法分离到的农药降解菌株存在稳定性差、底物谱窄、安全性不明等问题.Pseudomonas putida KT2440分离自根际土壤,是国际上公认的环境安全型菌株.利用同源重组方法将4种农药降解酶基因pytH、mpd、chd和amy A同时整合到P.putid... 详细信息

通过富集培养法分离到的农药降解菌株存在稳定性差、底物谱窄、安全性不明等问题.Pseudomonas putida KT2440分离自根际土壤,是国际上公认的环境安全型菌株.利用同源重组方法将4种农药降解酶基因pytH、mpd、chd和amy A同时整合到P.putida KT2440的染色体上,获得了一株无抗性基因残留的工程菌P.putida KT-pmca.工程菌KTpmca能够同时降解拟除虫菊酯类杀虫剂、有机磷类杀虫剂、百菌清和氯代酰胺类4种农药,百菌清降解率大于95%,甲氰菊酯、毒死蜱和敌稗的降解率均达到80%以上.此外,传代实验证明4种降解酶基因能够在遗传中稳定存在.本研究表明工程菌KT-pmca可为微生物原位修复消除农药污染提供新的菌株资源.

关键词：同源重组农药微生物降解 Pseudomonas putida KT2440 基因工程菌

一株促甲烷氧化假单胞菌Pseudomonas putida P7的分离及电活性特征

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微生物学报 2020年第9期60卷 2062-2071页

作者：郝钦钦张月超李新刘芳华中国科学院烟台海岸带研究所海岸带生物学与生物资源利用重点实验室山东烟台264003 中国科学院海岸带环境过程与生态修复重点实验室(烟台海岸带研究所) 山东烟台264003 中国科学院大学北京100049

【目的】解析一株从黄河三角洲湿地甲烷氧化富集物中分离获得的甲烷氧化菌伴生菌的生理学及电化学特性,并探究该菌株对甲烷氧化过程的影响。【方法】使用高通量测序技术解析甲烷氧化富集物的菌群结构,采用稀释涂布法、平板划线法分离甲... 详细信息

【目的】解析一株从黄河三角洲湿地甲烷氧化富集物中分离获得的甲烷氧化菌伴生菌的生理学及电化学特性,并探究该菌株对甲烷氧化过程的影响。【方法】使用高通量测序技术解析甲烷氧化富集物的菌群结构,采用稀释涂布法、平板划线法分离甲烷氧化菌的伴生菌,通过16S rRNA基因测序技术进行菌株初步鉴定。利用扫描电子显微镜观察菌株形态,并通过气相色谱(gas chromatography,GC)检测伴生菌利用甲烷情况及对甲烷氧化菌氧化甲烷效率的影响。采用双室微生物燃料电池(microbial fuel cells,MFCs)及差分脉冲伏安法(differential pulse voltammetry,DPV)检测菌株的电化学活性。【结果】黄河三角洲湿地土壤甲烷氧化富集物主要的好氧甲烷氧化菌为甲基杆菌属Methylobacter,同时还发现一些伴生菌。分离得到一株甲醇利用菌P7,其16S rRNA基因序列与恶臭假单胞菌Pseudomonas putida的相似性达99.79%。扫描电镜结果显示该菌株为杆状,长约1.5–2.5μm,宽度约为0.5μm。GC检测结果显示,该菌株不能利用甲烷,但与甲烷氧化菌共培养时,可以促进甲烷氧化(Pputida P7,该菌株可以促进甲烷氧化。本研究加深了对甲烷氧化过程中伴生菌的生理学特性及功能的认识。

关键词：甲烷氧化湿地 Pseudomonas putida Methylobacter 电化学活性

Polyhydroxyalkanoate Production by Pseudomonas putidaKT217 on a Condensed Corn Solubles Based Medium Fed with Glycerol Water or Sunflower Soapstock

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Advances in Microbiology 2012年第3期2卷 241-251页

作者： Jeremy Javers Chinnadurai Karunanithy Agricultural and Biosystems Engineering Department South Dakota State University Brookings USA Biology and Microbiology Department South Dakota State University Brookings USA

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.

关键词： Polyhydroxyalkanoate Pseudomonas putida Condensed Corn Solubles Glycerol Soapstock

Mass production, fermentation, formulation of Pseudomonas putidafor controlling of die back and phomopsis diseases on grapevine

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Advances in Bioscience and Biotechnology 2013年第6期4卷 741-750页

作者： Wafaa M. Haggag Malaka A. E. Saleh Inas Mostafa Noran Adel Biotechnology and Genetic Engineering Unit Dokki Egypt Pathology Department National Research Centre Dokki Egypt Pomology Department National Research Centre Dokki Egypt

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.

关键词： Die Back Phomopsis Diseases Grapevine and Pseudomonas putida

Zinc Oxide Nanoparticles in Bacterial Growth Medium: Optimized Dispersion and Growth Inhibition of Pseudomonas putida

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Advances in Nanoparticles 2013年第4期2卷 287-293页

作者： Manuela Vielkind Ingo Kampen Arno Kwade Institute for Particle Technology Braunschweig University of Technology Braunschweig Germany

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).

关键词： Bacterial Growth Medium Optimized Dispersion Pseudomonas putida Zinc Oxide Nanoparticles

Isolation and molecular characterization of a novel pseudomonas putidastrain capable of degrading organophosphate and aromatic compounds

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Advances in Biological Chemistry 2013年第6期3卷 564-578页

作者： Rupa Iyer Victor G. Stepanov Brian Iken Department of Biology and Biochemistry University of Houston Houston USA

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.

关键词： Organophosphates Aromatic Hydrocarbons Bioremediation Pseudomonas putida