锑氧化菌Pseudomonas sp.AO-1的分离鉴定及其对Sb(III)的氧化性能

采用抗性筛选法从锡矿山筛选出一株锑氧化菌,并利用分子生物学技术对其进行鉴定;考察了其氧化Sb()Ⅲ的性能和氧化次生矿物的特征.结果表明:锑氧化菌属于假单胞菌属(Pseudomonas),将其命名为Pseudomonas sp.AO-1(简称:AO-1);影响AO-1氧化Sb(Ⅲ)的因素主要有溶液pH值、溶解氧和铁锰氧化物(单质铁、FeCl_(3)和MnO_(2))等;AO-1在好氧和缺氧条件下均能氧化Sb(Ⅲ),好氧氧化Sb(Ⅲ)的米门常数Km和最大氧化速率Vmax值分别为393.05µmol/L和0.271µmol/(L·min),体现了较强的锑氧化性;AO-1和铁锰氧化物的耦合作用能促进Sb(Ⅲ)的氧化,且铁锰氧化物促进AO-1氧化Sb(Ⅲ)的速率依次为:FeCl_(3)>MnO_(2)>单质铁;AO-1和铁锰氧化物耦合氧化Sb(Ⅲ)生成含Sb(Ⅴ)的次生矿物,次生矿物会加速Sb(Ⅲ)的氧化以及影响锑在环境中的迁移转化.菌株AO-1的锑氧化性能良好,对于锑的生物化学转化和土壤微生物修复的应用有重要意义.

砷氧化菌Pseudomonas sp.AO-1的分离鉴定及其对As(Ⅲ)的氧化性能研究

随着环境砷污染日趋严重和不断地传播使得人类健康处于高风险之中,修复治理环境中的砷污染刻不容缓。将As(Ⅲ)氧化为As(Ⅴ)是治理砷污染的关键步骤,因此寻找一种经济且绿色的氧化技术成为了研究热点,而微生物氧化As(Ⅲ)被认为是治理砷污染一种绿色经济可行的方法。从湖南锡矿山矿区含砷的土壤中筛选出一株砷氧化菌,并利用分子生物学技术对其进行了鉴定;研究了其对As(Ⅲ)的抗性,不同As(Ⅲ)浓度下的生长特征曲线,不同pH条件下As(Ⅲ)的氧化性能,生长曲线及其氧化动力学。研究结果表明:该菌属于假单胞菌属(Pseudomonas),将其命名为Pseudomonas sp.AO-1(简称:AO-1),其对As(Ⅲ)的抗性达2000 mg·L^(-1),具有较高的抗性;当As(Ⅲ)质量浓度高于100 mg·L^(-1)时,菌株AO-1的生长受到了明显的抑制,其进入对数增殖期的时间延后,稳定期较短,衰亡期提前;pH对菌株AO-1的生长及As(Ⅲ)氧化率有显著的影响,其最适生长pH为7;当pH为3-8时,菌株AO-1氧化As(Ⅲ)的氧化率随pH的升高先提高后减小,pH为7时As(Ⅲ)氧化率到达最大的72.5%;菌株AO-1好氧化氧化As(Ⅲ)的米门常数Km和最大氧化速率Vmax值分别为274.70μmol·L^(-1)和0.31μmol·(L·min)-1,其Km低于一些已研究的菌株,体现菌株AO-1对As(Ⅲ)具有良好的亲和力,也表明了其具有较好的As(Ⅲ)氧化性能。综上,该研究所筛选的菌株AO-1具有良好的耐砷性和As(Ⅲ)氧化性,在水体及土壤砷污染修复与治理中具有的潜在应用价值。

Induction of Tolerance to Fusarium Wilt and Defense-Related Mechanisms in the Plantlets of Susceptible Berangan Banana Pre-Inoculated with Pseudomonas sp.(UPMP3) and Burkholderia sp.(UPMB3)

This study is aimed at assessing the ability of two endophytic bacteria originally isolated from healthy oil palm roots, Pseudomonas sp. （UPMP3） and Burkholderia sp. （UPMB3） to induce resistance in susceptible Berangan banana against Fusarium oxysporum race 4 （FocR4）. Increased accumulation of resistance-related enzymes such as peroxidase （PO）, phenylalanine ammonia lyase （PAL）, lignithioglycolic acid （LTGA）, and pathogenesis-related （PR） proteins （ehitinase and β-1,3-glucanase） has been observed in plantlets treated with endophytic bacteria UPMP3 and UPMB3 singly or as mixture under glasshouse conditions. Pre-inoculation of banana plantlets with UPMP3 showed a significant reduction in Fusarium wilt incidence 72 d after challenged inoculation with FocR4. UPMB3 was less effective in suppressing Fusarium wilt compared to UPMP3, whereas, the mixture of both endophytes showed an intermediate effect. Based on these results, it is concluded that UPMP3 could be a promising biological control agent that can trigger resistance against Fusarium wilt in susceptible Berangan banana.

Metabolism-independent chemotaxis of Pseudomonas sp. strain WBC-3 toward aromatic compounds

Pseudomonas sp. strain WBC-3 utilized methyl parathion or para-nitrophenol (PNP) as the sole source of carbon, nitrogen, and energy, and methyl parathion hydrolase had been previously characterized. Its chemotactic behaviors to aromatics were investigated. The results indicated that strain WBC-3 was attracted to multiple aromatic compounds, including metabolizable or transformable substrates PNP, 4-nitrocatechol, and hydroquinone. Disruption of PNP catabolic genes had no e?ect on its chemotactic behaviors w...

Optimization for Microbial Degradation of Dibenzothiophene by Pseudomonas sp. LKY-5 Using Response Surface Methodology

In this research, the degradation of dibenzothiophene(DBT) was investigated by using Pseudomonas sp. LKY-5 isolated from oil contaminated soil. The response surface methodology(RSM) based on the Box-Behnken design(BBD) was applied for evaluating the interactive effects of four independent variables including substrate concentration, temperature, pH and agitation rate on the DBT removal response. A total of 29 experiments for four factors at three levels were conducted in present study. A second-order regression model was then developed, and the analysis of variance(ANOVA) illustrated that the proposed quadratic model could be utilized to navigate the design space. The value of determination coefficient(R2=0.953 4) indicated a satisfactory agreement between the quadratic model and the experimental data. It was found that DBT removal was more significantly affected(P<0.000 1) by substrate concentration compared with other three parameters. An 100% degradation of DBT could be obtained by Pseudomonas sp. LKY-5 at a substrate concentration of 100 mg/L.

Pseudomonas sp.TK35-L enhances tobacco root development and growth by inducing HRGPnt3 expression in plant lateral root formation

Rhizosphere colonization is a key requirement for the application of plant growth-promoting rhizobacteria(PGPR)as a bioferilizer.Signaling molecules are often exchanged between PGPR and plants,and genes in plants may respond to the action of PGPR.Here,the luciferase luxAB gene was electrotransformed into Pseudomonas sp.strain TK35,a PGPR with an afinity for tobacco,and the labelled TK35(TK35-L)was used to monitor colonization dynamics in the tobacco rhizosphere and evaluate the effects of colonization on tobacco growth and root development.The transcript levels of the hydroxyproline rich glycoprotein HRGPnt3 gene,a lateral root induction indicator,in tobacco roots were examined by qPCR.The results showed that TK35-L could survive for long periods in the tobacco rhizosphere and colonize new spaces in the tobacco rhizosphere following tobacco root extension,exhibiting significant increases in root development,seedling growth and potassium accumulation in tobacco plants.The upregulation of HRGPnt3 transcription in the inoculated tobacco suggested that TK35-L can promote tobacco root development by upregulating the transcript levels of the HRGPnt3 gene,which promotes tobacco seedling growth.These findings lay a foundation for future studies on the molecular mechanism underlying the plant growth-promoting activities of PGPR.Futhermore,this work provided an ideal potential strain for biofertilizer production.

Extracellular synthesis of silver nanoparticles by Pseudomonas sp. THG-LS1.4 and their antimicrobial application

Silver nanoparticles（Ag NPs） are known to have bacteriostatic and bactericidal effects. The present study highlights the extracellular synthesis of Ag NPs and its antibacterial properties. The Ag NPs were synthesized using Pseudomonas sp. THG-LS1.4 strain which had been isolated from soil. The Ag NPs were characterized by field emission-transmission electron microscopy（FE-TEM）, X-ray diffraction（XRD）,Fourier transform-infrared（FT-IR） spectroscopy, and particle size distribution（DLS）. The Ag NPs displayed maximum absorbance at 412 nm and were irregular in shape ranging from 10 to 40 nm. The XRD spectroscopy results demonstrated the crystalline nature of nanoparticles. The Ag NPs showed antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Candida tropicalis, Vibrio parahaemolyticus, Escherichia coli and Pseudomonas aeruginosa. Furthermore, the Ag NPs were also evaluated for their increased antibacterial activities with various antibiotics against Escherichia coli, Pseudomonas aeruginosa and Salmonella enterica. Additionally, Ag NPs showd biofilm inhibition activity. The biosynthesized Ag NPs were found to be a potent agent against tested pathogens. More importantly, we highlight the applications of Ag NPs as an antimicrobial agent.

Purification of Pseudomonas sp. Lipase by Continuous Elution Electrophoresis Based on Pb^(2+) Precipitation Method

A Pb 2+ precipitation method was designed to get rid of the impure proteins in a lipase. The results show that it was a simple way in the primary treatment of the crude samples and about 20% impure proteins were removed in the precipitation step. Further, continuous elution electrophoresis was also applied as a preparative technique for attaining the highly pure lipase. During the continuous elution electrophoresis, the enzyme was eluted as a single peak and 5.7-fold purification was achieved in a yield of 54.3%. The two steps finally yielded an electrophoretically homogeneous enzyme.

Screening and pilot-scale evaluation of a highly efficient pesticide-degrading Pseudomonas sp.strain BL5

The widespread use of pesticides has caused serious harm to ecosystems,necessitating effective and environmentally friendly treatment methods.Bioremediation stands out as a promising approach for pollutant treatment,wherein the metabolic activities of microorganisms can transform toxic pesticides into compounds with lower or no toxicity.In this study,we obtained eight pesticide-degrading strains from pesticide-contaminated sites through continuous enrichment and screening.Four highly efficient pesticide-degrading strains(degradation ratios exceeding 80%)were identified.Among them,Pseudomonas sp.BL5 exhibited the strongest growth(exceeding 10^(9) CFU·ml^(-1))and outstanding degradation of benzene derivatives and chlorinated hydrocarbons at both laboratory and pilot scales,with degradation ratios exceeding 98%and 99.6%,respectively.This research provides new tools and insights for the bioremediation of pesticide-related pollutants.

Degradation of polycyclic aromatic hydrocarbons by Pseudomonas sp.JM2 isolated from active sewage sludge of chemical plant

It is important to screen strains that can decompose polycyclic aromatic hydrocarbons （PAHs） completely and rapidly with good adaptability for bioremedlation in a local area. A bacterial strain JM2, which uses pbenanthrene as its sole carbon source, was isolated from the active sewage sludge from a chemical plant in Jilin, China and identified as Pseudomonas based on 16S rDNA gene sequence analysis. Although the optimal growth conditions were determined to be pH 6.0 and 37~C, JM2 showed a broad pH and temperature profile. At pH 4.5 and 9.3, JM2 could degrade more than 40% of fluorene and phenanthrene （50 mg/L each） within 4 days. In addition, when the temperature was as low as 4~C, JM2 could degrade up to 24% fluorene and 12% phenanthrene. This showed the potential for JM2 to be applied in bioremediation over winter or in cold regions. Moreover, a nutrient augmentation study showed that adding formate into media could promote PAH degradation, while the supplement of salicylate had an inhibitive effect. Furthermore, in a metabolic pathway study, salicylate, phthalic acid, and 9-fluorenone were detected during the degradation of fluorene or phenanthrene. In conclusion, Pseudomonas sp. JM2 is a high performance strain in the degradation of fluorene and phenanthrene under extreme pH and temperature conditions. It might be useful in the bioremediation of PAHs.

Characterization and biodegradation kinetics of a new cold-adapted carbamazepine-degrading bacterium, Pseudomonas sp. CBZ-4

Carbamazepine is frequently detected in waters and hardly eliminated during conventional wastewater treatment processes due to its complicated chemical structure and resistance to biodegradation. A carbamazepine-degrading bacterium named CBZ-4 was isolated at a low temperature （10℃） from activated sludge in a municipal wastewater treatment plant. Strain CBZ-4, which can use carbamazepine as its sole source of carbon and energy, was identified as Pseudomonas sp. by the 16S rRNA gene sequence. The composition and percentage of fatty acids, which can reveal the cold-adaptation mechanism of strain CBZ-4, were determined. Strain CBZ-4 can effectively degrade carbamazepine at optimal conditions： pH 7.0, 10℃, 150 r/min rotation speed, and 13% inoculation volume. The average removal rate of carbamazepine was 46.6% after 144 hr of incubation. The biodegradation kinetics of carbamazepine by CBZ-4 was fitted via the Monod model. Vmax and Ks were found to be 0.0094 hr^-1 and 32.5 mg/L, respectively.

Pseudomonas sp.ZXY-1,a newly isolated and highly efficient atrazine-degrading bacterium,and optimization of biodegradation using response surface methodology

Atrazine, a widely used herbicide, is increasing the agricultural production effectively, while also causing great environmental concern. Efficient atrazine-degrading bacterium is necessary to removal atrazine rapidly to keep a safe environment. In the present study, a new atrazine-degrading strain ZXY-1, identified as Pseudomonas, was isolated. This new isolated strain has a strong ability to biodegrade atrazine with a high efficiency of 9.09 mg/L/hr.Temperature, p H, inoculum size and initial atrazine concentration were examined to further optimize the degradation of atrazine, and the synthetic effect of these factors were investigated by the response surface methodology. With a high quadratic polynomial mathematical model（R^2= 0.9821） being obtained, the highest biodegradation efficiency of 19.03 mg/L/hr was reached compared to previous reports under the optimal conditions（30.71°C, pH 7.14, 4.23%（V/V） inoculum size and 157.1 mg/L initial atrazine concentration）.Overall, this study provided an efficient bacterium and approach that could be potentially useful for the bioremediation of wastewater containing atrazine.

Cloning,expression and characterization of L-cysteine desulfhydrase gene from Pseudomonas sp.TS1138

L-cysteine desulfhydrase(CD)plays an impor-tant role in L-cysteine decomposition.To identify the CD gene in Pseudomonas sp.TS1138 and investigate its effect on the L-cysteine biosynthetic pathway,the CD gene was cloned from Pseudomonas sp.TS1138 by polymerase chain reaction(PCR)method.The nucleotide sequence of CD gene was determined to be 1,215 bp,and its homology with other sequences encoding CD was analyzed.Then the CD gene was subcloned into pET-21a(+)vector and expressed in Escherichia coli(E.coli)by isopropyl-β-D-thiogalacto-pyranoside(IPTG)inducement.The recombinant CD was purified by Ni-NTA His-Bind resin,and its activity was identified by the CD activity staining.The enzymatic proper-ties of the recombinant CD were characterized and its critical role involved in the L-cysteine biosynthetic pathway was also discussed.

Phosphate-Solubilizing and -Mineralizing Abilities of Bacteria Isolated from Soils

Microorganisms capable of solubilizing and mineralizing phosphorus （P） pools in soils are considered vital in promoting P bioavallability. The study was conducted to screen and isolate inorganic P-solubilizing bacteria （IPSB） and organic P-mineralizing bacteria （OPMB） in soils taken from subtropical flooded and temperate non-flooded soils, and to compare inorganic P-solubilizing and organic P-solubilizing abilities between IPSB and OPMB. Ten OPMB strains were isolated and identified as Bacillus cereus and Bacillus megaterium, and five IPSB strains as B. megaterium, Burkholderia caryophyUi, Pseudomonas cichorii, and Pseudomonas syringae. P-solubilizing and -mineralizing abilities of the strains were measured using the methods taking cellular P into account. The IPSB strains exhibited inorganic P-solubilizing abilities ranging between 25.4-41.7 μg P mL^-1 and organic P-mineralizing abilities between 8.2-17.8μg P mL^-1. Each of the OPMB strains also exhibited both solubilizing and mineralizing abilities varying from 4.4 to 26.5 μg P mL^-1 and from 13.8 to 62.8 μg P mL^-1, respectively. For both IPSB and OPMB strains, most of the P mineralized from the organic P source was incorporated into the bacterial cells as cellular P. A significantly negative linear correlation （P 〈 0.05） was found between culture pH and P solubilized from inorganic P by OPMB strains. The results suggested that P solubilization and mineralization could coexist in the same bacterial strain.

Decolorization and bio degradation metabolism of azo dyes by Pseudomonas S-42

A bacterial strain was isolated from activated sludge and has been identified as Pseudomonas sp. S-42 capable of decolorizing azo dyes such as Diamira Brilliant Orange RR (DBO-RR), Direct Brown M (DBM), Eriochrome Brown R (EBR) and so on. The growing cells, intact cells, cell-free extract and purified enzyme of strain S-42 could decolorize azo dyes under similar conditions at the optimum pH 7.0 and temperature of 37℃. The efficiencies of decolorization for DBO-RR, DBM, EBR with intact cells stood more than 90%. When the cell concentration was 15mg (wet)/ml and the reaction time was 5 hours, the decolorizing activities of intact cells for above three azo dyes were 1.75, 2.4, 0.95 μg dye/mg cell, respectively. Cell-free extract and purified enzyme belonged to azoreductase with molecular weight about 34000±2000 and Vmax and Km values for DBO-RR of 13μmol/mg protein/nun and 54μmol, respectively. The results from the detection of the biodegradation products of DBO-RR by spectrophotometric and NaNO2 reaction methods showed that the biodegradation of azo dyes was initiated by the reducing cleavage of azo bonds. The biodegradation metabolism path for DBO-RR by Psued. S-42 was hypothesized.

Antibiogram of Escherichia coli and Pseudomonas Strains Isolated from Wastewater Generated by an Abattoir as It Journeys into a Receiving River

Untreated wastewater from abattoir operations contains nutrients and other components that aid the growth of microorganisms especially bacteria. They also serve as a habitat for potentially pathogenic bacteria which might be a source of public health concern. The study was carried out to determine the antibiotics susceptibility profile of Gram-negative bacteria (Pseudomonas and Escherichia coli) to selected antibiotics. Wastewater samples were collected from ten different sampling points and cultured on Eosin Methylene Blue (EMB) and King’s B medium. The bacterial strains obtained from the wastewater samples were subjected to antibiotics susceptibility tests, using the disc diffusion technique. A total of 60 Pseudomonas and 100 Escherichia coli were isolated out of which none of the Pseudomonas strains showed resistance to imipenem, colistin sulphate, meropenem and aztreonam, while 100% resistance was observed to ceftazidime and piperacillin. All the Escherichia coli strains were resistant to oxacillin and ceftazidime, while the percentage resistance to aztreonam, ertapenem, cefoxitin and tetracyline was 6%, 11%, 43% and 58% respectively. Eighty-five percent (85%) of the total Escherichia coli showed resistance to more than two antibiotics, while 14% showed resistance to ceftazidime and oxacillin, with only one isolate showing resistance to ceftazidime and cefoxitin. There is the need for an effective treatment of wastewater generated from abattoir operations to prevent the potential spread and transmission of antibiotic resistant bacteria to the human population who depends heavily on some of the water bodies, receiving input from abattoir wastes.

Studies on Bioflocculant Production by Pseudoalteromonas sp. NUM8, a Marine Bacteria Isolated from the Circulating Seawater

A bioflocculant producing potential bacteria was isolated from the circulating seawater of bio-filter using streak plate methods.The bacteria was identified through biochemical characteristics,partial 16S ribosomal ribonucleic acids(rRNA),nucleo-tide sequencing,and BLAST analysis of the gene sequence that showed the bacteria have 99%similarity to Pseudoalteromonas sp.and deposited in GenBank as Pseudoalteromonas sp.NUM8 with accession number JX435820.Influences of time course assay,carbon sources,nitrogen sources,inoculum size,as well as initial pH on the bacteria producing extracellular bioflocculant activity were investigated.The results showed that the strain optimal production period of microbial bioflocculant was at 72 h(flocculating activity of 94.5%),then dropped slowly.The bacteria optimally produced the bioflocculant when 1.0%sucrose and 0.5%sodium nitrate were used as sole sources of carbon and nitrogen with flocculating activities of 92.8%and 93.8%respectively.Also,the bacteria produced the bioflocculant optimally when initial pH of the medium was 5.0(flocculating activity 93.2%),and when Ca^(2+)was used as cation(flocculating activity 93.4%).The culture condition of inoculum size of 3%(v/v)was optimal flocculant pro-duction(flocculating activity 94.4%).Composition analyses indicated the bioflocculant to be principally a glycoprotein made up of about 34.3%protein and 63.4%total carbohydrate.

Isolation, characterization and phylogenetic analysis of a bacterial strain capable of degrading acetamiprid

An aerobic bacterium, capable of degrading the new chloronicotine pesticide acetamiprid, was isolated from the sludge of pesticide factory after successive enrichment cultures and named strain FH2 which is a Gram-negative, rod-shaped, obligate aerobic organism with （（0.5-0.7）×（l.5-3.0））μm of cell size and with monotrichous flagellum. It was identified as a member of Pseudomonas sp. based on morphology, physio-biochemical properties, Biolog GN2, 16S rDNA sequence and phylogenetic characteristic analysis. The isolate could grow optimally at pH 7.0 and 30℃ in acetamiprid-mineral medium with 800 mg/L concentration. About 53.3% acetamiprid was degraded by strain FH2 after incubation for 14 d in acetamiprid-mineral medium and nearly 96.7% degraded when incubated in acetamiprid-yeast mineral medium at 30℃ for 14 d. This paper describes phylogenetic and degradation characterization of a pure bacterium being able to mineralize acetamiprid for the first time.

Biodegradation Kinetics and Mechanism of a Typical Coal Gasification Pollutant:Quinoline

A strain Pseudomonas sp.PP1 was isolated from activated sludge and characterized by morphological observation,biochemical and physiological identification as well as 16 S rRNA gene sequence analysis.It was a gram-negative,non-motile,rod-shaped with colonies which were white,opaque,small and wrinkled on solid quinoline-MSM.The strain was negative for catalase,gelatin liquefaction,M.R and V.P tests and was unable to reduce / restore nitrate.The 1406 bp 16 S rRNA gene fragment of PP1 was more than 99% identical to the Pseudomonas sp.Biodegradation of quinoline as single substrate was conducted in batch experiments at different temperatures( 15- 40 ℃) and p H values( 5- 10).The results indicate that the optimum conditions for the degradation of quinoline by PP1 were 30 ℃ and p H 7.Pseudomonas sp.PP1 was able to degrade 97.1%,95.4%,94.8%,63.6% and 40.4% quinoline when initial concentrations of quinoline were 50,100,200,300,and 400 mg / L,respectively,and lag phases were prolonged from 2 h to 10 h.The maximum degradation rate( q) was obtained at an initial quinoline concentration( S0) of about 100 mg/L with q of 0.082.The experimentally obtained q values at various initial S0 were fitted by Haldane model,Yano model,Aiba model,Edward model and Webb model,and the results demonstrated that Haldane model gives the best fit for strain PP1 with coefficient of determination,R2= 0.9124 and SDavg= 0.0113208.Five metabolic intermediates were identified by high performance liquid chromatograph( HPLC) and GC / MS.Finally,a possible pathway containing 5,6-dihydroxy-2-oxo-1,2-dihydroquinoline as an intermediate was proposed for the first time.

A Test for Stabilization of an Oligomeric Protein by Introduction of Aromatic Residues into the Interface

The design of variants to enhance conformational stability of proteins is an important aspect of protein engineering. Oligomeric proteins are often stabilized by aromatic clusters located within the subunit interfaces. In the present study, the authors constructed five variants of Ps3aHSD （Pseudomonas sp. B-0831 3a-hydroxysteroid dehydrogenase） in which one or two residues at the dimer interface were replaced with aromatic residues, and examined the effects of introducing aromatic residues in this region on protein thermostability. Under their experimental conditions, all variants formed dimers, similar to wild-type Ps3aHSD. Thermal denaturation experiments indicated that Tm of all variants was 0.2-16.2 °C lower than that of wild-type protein, indicating less stable thanwild-type protein. The results collectively suggest that aromatic residues of natural oligomeric proteins are strictly posted in the interface to facilitate optimal interactions and avoid conformational strain.