DOCSLIB.ORG

Cellular and Transcriptional Response of Pseudomonas Stutzeri to Quantum Dots Under Aerobic and Denitrifying Conditions † ‡ ‡ † Yu Yang, Huiguang Zhu, Vicki L

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cellular and Transcriptional Response of Pseudomonas Stutzeri to Quantum Dots Under Aerobic and Denitrifying Conditions † ‡ ‡ † Yu Yang, Huiguang Zhu, Vicki L ARTICLE pubs.acs.org/est Cellular and Transcriptional Response of Pseudomonas stutzeri to Quantum Dots under Aerobic and Denitrifying Conditions † ‡ ‡ † Yu Yang, Huiguang Zhu, Vicki L. Colvin, and Pedro J. Alvarez*, †Department of Civil and Environmental Engineering, Rice University, Houston, Texas 77005, United States ‡Department of Chemistry, Rice University, Houston, Texas 77005, United States bS Supporting Information ABSTRACT: Pseudomonas stutzeri was exposed to quantum dots (QDs) with three different surface coatings (anionic polymaleic anhydride-alt-1-octadecene (PMAO), cationic polyethylenimine (PEI), and carboxyl QDs) under both aerobic and anaerobic (denitrifying) conditions. Under aerobic conditions, toxicity (assessed per growth inhibition) increased from PMAO to carboxyl to PEI QDs. The positive charge of PEI facilitated direct contact with negatively charged bacteria, which was verified by TEM analysis. Both PMAO and PEI QDs hindered energy transduction (indicated by a decrease in cell membrane potential), and this effect was most pronounced with PEI QDs under denitrifying conditions. Up- regulation of denitrification genes (i.e., nitrate reductase narG, periplasmic nitrate reductase napB, nitrite reductase nirH, and NO reductase norB) occurred upon exposure to subinhibitory PEI QD concentrations (1 nM). Accordingly, denitri- fication activity (assessed per respiratory nitrate consumption in the presence of ammonia) increased during sublethal PEI QD exposure. However, cell viability (including denitrification) was hindered at 10 nM or higher PEI QD concentra- tions. Efflux pump genes czcB and czcC were induced by PEI QDs under denitrifying conditions, even though Cd and Se dissolution from QDs did not reach toxic levels (exposure was at pH 7 to minimize hydrolysis of QD coatings and the associated release of metal constituents). Up- regulation of the superoxide dismutase (stress) gene sodB occurred only under aerobic conditions, likely due to intracellular production of reactive oxygen species (ROS). The absence of ROS under denitrifying conditions suggests that the antibacterial activity of QDs was not due to ROS production alone. Overall, this work forewarns about unintended potential impacts to denitrification as a result of disposal and incidental releases of QDs, especially those with positively charged coatings (e.g., PEI QDs). ’ INTRODUCTION water pollutants) coupled with the production of dinitrogen gas 12À14 More than one-half century has passed since Richard Feynman (N2) through intermediate nitrogen oxides (NO and N2O). presented the perspective of nanomaterial manipulation and Consequently, denitrification results in a loss of nitrogen 1 (a common limiting nutrient) from aquatic and soil systems with nanotechnology application. Nanotechnology is now leading a 15 concomitant production of a greenhouse gas (N2O). Pseudo- creative and epochal revolution covering various subjects, such as 16,17 pharmaceutics, electronics, physics, chemistry, and biology.2À4 monas stutzeri is one of the most studied denitrifying bacteria, and its denitrification genes have been discerned and sequenced With the increasing worldwide usage of manufactured nano- 17 materials (MNMs), their accidental or incidental releases to (Figure 1). Thus, P. stutzeri is a convenient model bacterium to the environmental seem inevitable. Accordingly, the potential investigate the potential impact of MNMs to denitrification adverse effect of MNMs to different types of biological molecules, processes and associated bacterial response, which has not yet cells, and organisms has received significant recent attention.5À11 been addressed in the literature. However, considerable uncertainty remains about the potential Quantum dots (QDs) offer valuable functionality for bioimag- impacts to microbial ecosystem services (e.g., nutrient cycling) ing, solar cells, electronics, and drug (or gene) delivery, since these semiconductor nanocrystals possess uniform sizes and and the associated risks to ecosystem health. 2,18À20 The nitrogen cycle is one of the fundamental biogeochemical distinct optical and electrical properties. Some QDs can processes on the earth and denitrification is a key component that is critically implicated in water quality, agricultural productivity, Received: December 20, 2010 and climate change. This form of anaerobic respiration (which is Accepted: April 15, 2011 performed by a wide range of Bacteria and Archaea) involves the Revised: April 11, 2011 À À reduction of nitrate (NO3 ) or nitrite (NO2 ) (both common Published: April 28, 2011 r 2011 American Chemical Society 4988 dx.doi.org/10.1021/es1042673 | Environ. Sci. Technol. 2011, 45, 4988–4994 Environmental Science & Technology ARTICLE Random primers, dNTP set, Superscript II reverse transcriptase, Bacteria Counting Kit, BacLight Bacterial Membrane Potential Kit, and BacLight RedoxSensor Green Vitality Kit. RNAprotect Figure 1. Genes coding for denitrification in Pseudomonas stutzeri. The bacteria reagent, QIAquick PCR Purification Kit, and RNeasy effect of QDs on the genes depicted in bold was assessed by RT-qPCR. Mini Kit were obtained from Qiagen Inc. (Valencia, CA). SYBR Green Master Mix was purchased from Applied Biosystems be toxic to microorganisms and animals, and their toxicity (Carlsbad, CA), and PCR primers were synthesized by Integrated depends on their physiochemical properties such as core com- DNA Technologies, Inc. (San Diego, CA). Lysozyme, ethylene- position, size, charge, and surface coatings,6,8,21À24 which are diamine-tetraacetic acid (EDTA) buffer, Tris-EDTA (TE) buffer, critical determinants of QD stability and biosafety.25,26 Previous phosphate buffered saline (PBS), multielement standard solution, research showed that the chemical composition and stability of and other chemicals were provided by Sigma-Aldrich (St. Louis, the organic coating play a key role in cytotoxicity of QDs, MO). CHEMets colorimetric dissolved oxygen test kit was influencing the release of toxic heavy metal ions and the potential purchased from CheMetrics, Inc. (Calverton, VA). 25,27,28 uptake of QDs. Effect of QDs on Bacterial Growth. P. stutzeri was exposed to In this study we address the effect of three types of QDs on PEI QDs (1À80 nM) or PMAO QDs (50À800 nM) under P. stutzeri. Exposure was conducted in minimal mineral medium either aerobic or anaerobic conditions for 24 h at 37 °C. Higher under neutral pH conditions to minimize extensive weathering concentrations (0.5 and 1 μM) were used for carboxyl QDs, (e.g., hydrolysis) of the QD coating and avoid confounding which were less toxic; these tests were conducted under aerobic effects on bacterial toxicity associated with the release of Cd and conditions. Bacterial growth was monitored using flow cytome- Se ions.26 Bacterial response is assessed at cellular and molecular try, according to the Bacteria Counting Kit instructions, using a levels, under both anaerobic (denitrifying) and aerobic condi- BD LSRFortessa cell analyzer (Franklin Lakes, NJ). The experi- tions. Anaerobic conditions accentuated the susceptibility of ments were performed at least in triplicate and calculation of half- P. stutzeri to the toxic effect of the positively charged PEI QDs, maximal inhibitory concentration (IC50) was described in Sup- indicated by decreases in cell viability, cell membrane potential, porting Information (SI). Experiments were repeated for con- and reductase activity. We also demonstrated the up-regulation firmation purposes. of denitrification genes as a result of sublethal exposure, which is To assess the role of released heavy metal ions in microbial a novel finding with implications on nitrogen cycling. toxicity, the supernatant was collected by ultracentrifugation (35 000 rpm for 3.5 h) and Cd and Zn concentrations were ’ MATERIALS AND METHODS measured by inductively coupled plasma-mass spectrometry (ICP-MS) Elan 9000 (Perkin-Elmer, Waltham, MA). Then, QD Preparation and Characterization. Three different types additional growth inhibition tests were conducted by exposing of QDs were used in this work. CdSe/CdZnS quantum dots were P. stutzeri for 24 h to similar concentrations of heavy metal ions 29 synthesized as described earlier. Briefly, a mixture of purified (without QDs). The heavy metals were added as ZnCl2 and CdSe nanocrystal chloroform solution, octadecylamine (ODA), Cd(NO3)2 salts under both aerobic and anaerobic conditions. zinc/cadmium precursor, and thiourea/ethanol solution was The tested concentrations were 2 ppm Zn2þ and 0.2 ppm Cd2þ, stirred and heated to 150 °C, cooled to 60 °C, and separated 1 ppm Zn2þ and 0.1 ppm Cd2þ, and 0.5 ppm Zn2þ and 0.05 ppm by precipitation. Polyanionic polymaleic anhydride-alt-1-octade- Cd2þ. Growth was monitored by measuring optical density at cene (PMAO, Mn = 30 000À50 000) and polycationic polyethy- 600 nm with a SpectraMax plus spectrometer (Molecular Device, lenimine (PEI, Mn = 10 000) were used to transfer the as- Sunnyvale, CA). synthesized QDs into aqueous phase (Figure S1).30,31 Qdot Assessment of Membrane Potential and Redox Activity. 655 ITK carboxyl QDs were purchased from Invitrogen Inc. P. stutzeri was grown aerobically or anaerobically on Difco (Carlsbad, CA). Their size distribution and zeta-potential were nutrient broth at 37 °C overnight, and diluted to an OD600 of measured in the exposure medium using a Zetasizer Nano 0.002 in PBS buffer.35 The cells were exposed to 0.5 μM PEI or (Malvern Instruments, U.K.), and concentrations were calcu- PMAO QDs for 15 min. The BacLight Bacterial
Load more

Recommended publications
  • Aerobic and Anaerobic Bacterial and Fungal Degradation of Pyrene: Mechanism Pathway Including Biochemical Reaction and Catabolic Genes
    International Journal of Molecular Sciences Review Aerobic and Anaerobic Bacterial and Fungal Degradation of Pyrene: Mechanism Pathway Including Biochemical Reaction and Catabolic Genes Ali Mohamed Elyamine 1,2 , Jie Kan 1, Shanshan Meng 1, Peng Tao 1, Hui Wang 1 and Zhong Hu 1,* 1 Key Laboratory of Resources and Environmental Microbiology, Department of Biology, Shantou University, Shantou 515063, China; [email protected] (A.M.E.); [email protected] (J.K.); [email protected] (S.M.); [email protected] (P.T.); [email protected] (H.W.) 2 Department of Life Science, Faculty of Science and Technology, University of Comoros, Moroni 269, Comoros * Correspondence: [email protected] Abstract: Microbial biodegradation is one of the acceptable technologies to remediate and control the pollution by polycyclic aromatic hydrocarbon (PAH). Several bacteria, fungi, and cyanobacteria strains have been isolated and used for bioremediation purpose. This review paper is intended to provide key information on the various steps and actors involved in the bacterial and fungal aerobic and anaerobic degradation of pyrene, a high molecular weight PAH, including catabolic genes and enzymes, in order to expand our understanding on pyrene degradation. The aerobic degradation pathway by Mycobacterium vanbaalenii PRY-1 and Mycobactetrium sp. KMS and the anaerobic one, by the facultative bacteria anaerobe Pseudomonas sp. JP1 and Klebsiella sp. LZ6 are reviewed and presented, to describe the complete and integrated degradation mechanism pathway of pyrene. The different microbial strains with the ability to degrade pyrene are listed, and the degradation of Citation: Elyamine, A.M.; Kan, J.; pyrene by consortium is also discussed.
    [Show full text]
  • Pseudomonas Chloritidismutans Sp. Nov., a Non- Denitrifying, Chlorate-Reducing Bacterium
    International Journal of Systematic and Evolutionary Microbiology (2002), 52, 2183–2190 DOI: 10.1099/ijs.0.02102-0 Pseudomonas chloritidismutans sp. nov., a non- denitrifying, chlorate-reducing bacterium Laboratory of Microbiology, A. F. W. M. Wolterink, A. B. Jonker, S. W. M. Kengen and A. J. M. Stams Wageningen University, H. van Suchtelenweg 4, 6703 CT Wageningen, Author for correspondence: The Netherlands A. F. W. M. Wolterink. Tel: j31 317 484099. Fax: j31 317 483829. e-mail: Arthur.Wolterink!algemeen.micr.wag-ur.nl A Gram-negative, facultatively anaerobic, rod-shaped, dissimilatory chlorate- reducing bacterium, strain AW-1T, was isolated from biomass of an anaerobic chlorate-reducing bioreactor. Phylogenetic analysis of the 16S rDNA sequence showed 100% sequence similarity to Pseudomonas stutzeri DSM 50227 and 986% sequence similarity to the type strain of P. stutzeri (DSM 5190T). The species P. stutzeri possesses a high degree of genotypic and phenotypic heterogeneity. Therefore, eight genomic groups, termed genomovars, have been proposed based upon ∆Tm values, which were used to evaluate the quality of the pairing within heteroduplexes formed by DNA–DNA hybridization. In this study, DNA–DNA hybridization between strain AW-1T and P. stutzeri strains DSM 50227 and DSM 5190T revealed respectively 805 and 565% similarity. DNA–DNA hybridization between P. stutzeri strains DSM 50227 and DSM 5190T revealed 484% similarity. DNA–DNA hybridization indicated that strain AW-1T is not related at the species level to the type strain of P. stutzeri. However, strain AW-1T and P. stutzeri DSM 50227 are related at the species level. The physiological and biochemical properties of strain AW-1T and the two P.
    [Show full text]
  • Section 4. Guidance Document on Horizontal Gene Transfer Between Bacteria
    306 - PART 2. DOCUMENTS ON MICRO-ORGANISMS Section 4. Guidance document on horizontal gene transfer between bacteria 1. Introduction Horizontal gene transfer (HGT) 1 refers to the stable transfer of genetic material from one organism to another without reproduction. The significance of horizontal gene transfer was first recognised when evidence was found for ‘infectious heredity’ of multiple antibiotic resistance to pathogens (Watanabe, 1963). The assumed importance of HGT has changed several times (Doolittle et al., 2003) but there is general agreement now that HGT is a major, if not the dominant, force in bacterial evolution. Massive gene exchanges in completely sequenced genomes were discovered by deviant composition, anomalous phylogenetic distribution, great similarity of genes from distantly related species, and incongruent phylogenetic trees (Ochman et al., 2000; Koonin et al., 2001; Jain et al., 2002; Doolittle et al., 2003; Kurland et al., 2003; Philippe and Douady, 2003). There is also much evidence now for HGT by mobile genetic elements (MGEs) being an ongoing process that plays a primary role in the ecological adaptation of prokaryotes. Well documented is the example of the dissemination of antibiotic resistance genes by HGT that allowed bacterial populations to rapidly adapt to a strong selective pressure by agronomically and medically used antibiotics (Tschäpe, 1994; Witte, 1998; Mazel and Davies, 1999). MGEs shape bacterial genomes, promote intra-species variability and distribute genes between distantly related bacterial genera. Horizontal gene transfer (HGT) between bacteria is driven by three major processes: transformation (the uptake of free DNA), transduction (gene transfer mediated by bacteriophages) and conjugation (gene transfer by means of plasmids or conjugative and integrated elements).
    [Show full text]
  • Effect of Toluene on Pseudomonas Stutzeri ST-9 Morphology: Plasmolysis, Cell Size and 1
    Canadian Journal of Microbiology Effect of toluene on Pseudomonas stutzeri ST -9 morphology: Plasmolysis, cell size and formation of outer membrane vesicles Journal: Canadian Journal of Microbiology Manuscript ID cjm-2016-0099.R1 Manuscript Type: Article Date Submitted by the Author: 18-Mar-2016 Complete List of Authors: Michael, Esti; Ariel University, Chemical Engineering Nitzan, YeshayahuDraft ; Bar-Ilan University Langzam, Yakov; Bar-Ilan University, The Mina & Everard Goodman Faculty of Life Sciences Luboshits, Galia ; Ariel University, Cahan, Rivka; Ariel University, Chemical Engineering Keyword: Pseudomonas, plasmolysis, toluene, outer membrane vesicles, morphology https://mc06.manuscriptcentral.com/cjm-pubs Page 1 of 27 Canadian Journal of Microbiology Effect of toluene on Pseudomonas stutzeri ST-9 morphology: Plasmolysis, cell size and 1 formation of outer membrane vesicles 2 3 Esti Michael 1,2 , Yeshayahu Nitzan 2, Yakov Langzam 2, Galia Luboshits 1 and Rivka Cahan 1* 4 1Department of Chemical Engineering, Ariel University, Ariel 40700, Israel 5 2The Mina & Everard Goodman Faculty of Life Sciences, Bar-Ilan University, 6 Ramat-Gan 52900, Israel 7 *Corresponding author 8 9 10 11 Draft 12 13 14 15 16 17 18 19 20 21 22 23 24 25 1 https://mc06.manuscriptcentral.com/cjm-pubs Canadian Journal of Microbiology Page 2 of 27 Abstract 1 Isolated toluene-degrading Pseudomonas stutzeri ST-9 bacteria were grown in a minimal 2 medium containing toluene (100 mg L -1) (MMT) or glucose (MMG) as the sole carbon source, 3 with specific growth rates of 0.019 h -1 and 0.042 h -1, respectively. Scanning (SEM) as well as 4 transmission (TEM) electron microscope analyses showed that the bacterial cells grown to mid 5 log in the presence of toluene possess a plasmolysis space.
    [Show full text]
  • Aerobic and Nitrate Respiration Routes of Carbohydrate Catabolism in Pseudomonas S Tut Zeri
    AN ABSTRACT OF THE THESIS OF WILLIAM JAN SPANGLER for the Ph. D. in MICROBIOLOGY (Name) (Degree) (Major) thesis is presented Date 7 / `> /q6! Title AEROBIC AND NITRATE RESPIRATION ROUTES OF CAR- BOHYDRATE CATABOLISM IN PSEUDOMONAS STUTZERI Abstract approved ( Major professor) Pseudomonas stutzeri and other denitrifying bacteria are able to grow under anaerobic conditions, using nitrate -oxygen as the terminal hydrogen acceptor, in a manner analagous to classical aerobic respiration with free -molecular oxygen. This rather unique phenomenon is known as nitrate respiration. Nitrate respiration has been studied with respect to the nitrate reducing enzymes and carrier systems involved in the reduction sequence, but very little emphasis has been placed on the metabolic pathways which are associated with nitrate respiration. This study was carried out in an attempt to establish the metabolic pathways operative, both under aerobic con- ditions and during nitrate respiration, in order to determine whether there was any shift of pathways under conditions of nitrate respira- tion. Primary pathways were determined by the radiorespirometric method using specifically labelled glucose and gluconate. The results, based primarily on the rate of decarboxylation of the C -1 and C -4 positions of glucose, indicated the operation of the Entner- Doudoroff and pentose phosphate pathways under both aerobic condi- tions and conditions of nitrate respiration. Evolution of 14CO2 from the other labels of glucose, as well as incorporation of these labels into the cell, indicated that terminal pathways such as the tricar- boxylic acid cycle or glyoxalate cycle might also be operative under both conditions of oxygen relationship. The secondary pathways were studied using specifically labelled acetate.
    [Show full text]
  • Isolation and Characterization of a Novel Denitrifying Bacterium with High Nitrate Removal: Pseudomonas Stutzeri
    Iran. J. Environ. Health. Sci. Eng., 2010, Vol. 7, No. 4, pp. 313-318 ISOLATION AND CHARACTERIZATION OF A NOVEL DENITRIFYING BACTERIUM WITH HIGH NITRATE REMOVAL: PSEUDOMONAS STUTZERI *1A. Rezaee, 2H. Godini, 1S. Dehestani, 1S. Kaviani 1 Department of Environmental Health, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2 Department of Environmental Health, School of Public Health, Lorestan University of Medical Sciences, Khoramabbad, Iran Received 16 August 2009; revised 13 Jully 2010; accepted 20 August 2010 ABSTRACT The aim of this study was to isolate and characterize a high efficiency denitrifier bacterium for reducing nitrate in wastewater. Six denitrifier bacteria with nitrate removal activities were isolated from a petrochemical industry effluent with high salinity and high nitrogen concentrations without treatment. The isolated bacteria were tested for nitrate reomoval activity. One of the bacterium displayed the highest reduction of nitrate. The strain was preliminarily identified using biochemical tests and further identified based on similarity of PCR-16S rRNA using universal primers. Biochemical and molecular experiments showed that the best bacterium with high nitrate removal potential was Pseudomonas stutzeri, a member of the α subclass of the class Proteobacteria. The extent of nitrate removal efficiency was 99% at 200 mg/L NO3 and the nitrite content of the effluent was in the prescribed limit. The experiments showed the ability of Pseudomonas stutzeri to rapidly remove nitrate under anoxic conditions. The strain showed to be potentially good candidate for biodenitrification of high nitrate solutions. Key words: Pseudomonas stutzeri; Denitrification; Polymerase Chain Reaction, Isolation; Characterization INTRODUCTION Biological denitrification is a process carried to respire anaerobically using nitrogen oxides as out by numerous genera of bacteria.
    [Show full text]
  • Characterization of Biosurfactant from Pseudomonas Stutzeri SJ3 for Remediation of Crude Oil-Contaminated Soil
    Characterization of Biosurfactant From Pseudomonas Stutzeri SJ3 for Remediation of Crude Oil-Contaminated Soil Sekar Harikrishnan ( [email protected] ) Annamalai University Faculty of Marine Sciences https://orcid.org/0000-0001-5139-6648 Singaram Jayalakshmi Annamalai University Faculty of Marine Sciences Mohamad S. Alsalhi King Saud University Alager Kartick Annamalai University Faculty of Marine Sciences Sandhanasamy Devanesan King Saud University Aruliah Rajasekar Thiruvalluvar University Research Article Keywords: Oil degrading bacteria, hydrocarbonoclastic bacteria, FTIR, Emulsication activity, Antibacterial activity Posted Date: June 2nd, 2021 DOI: https://doi.org/10.21203/rs.3.rs-497731/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/14 Abstract In the present work, production of biosurfactant was studied from the bacterial strains isolated from the soil samples collected from oil contaminated sites in Karaikal ONGC, Puducherry, India. Six morphologically different hydrocarbonoclastic bacterial strains (SJ1-SJ6) isolated on oil agar plates were further screened for biosurfactant production. Based on the screening methods results of 26 mm oil displacement zone, positive results of drop collapse test, 68.14% emulsication index (E24) and 79.2% of bacterial adherence percentage, the isolate SJ3 was selected as the most potent strain and it was identied as P. stutzeri using standard biochemical and 16S rRNA gene sequencing-based methods. Optimization of the P. stutzeri strain showed 36 h incubation, 150 rpm agitation, pH 7.5, 37oC, 1% salinity, 2% glucose as carbon source and 1% yeast extract as nitrogen source were the ideal conditions for growth and the biosurfactant production was found to be growth dependent.
    [Show full text]
  • Extracellular Electron Transfer of Pseudomonas Stutzeri Driven By
    Received: January 13, 2016 Electrochemistry Accepted: March 3, 2016 Published: May 5, 2016 The Electrochemical Society of Japan http://dx.doi.org/10.5796/electrochemistry.84.312 Communication Electrochemistry, 84(5), 312–314 (2016) Extracellular Electron Transfer of Pseudomonas stutzeri Driven by Lithotrophic and Mixotrophic Denitrification Tetsuya YAMADA,a Satoshi KAWAICHI,b Akihisa MATSUYAMA,c Minoru YOSHIDA,c Nobuhiro MATSUSHITA,d and Ryuhei NAKAMURAb,* a Department of Electronic Chemistry, Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute of Technology, 4259 Midori, Nagatsuta, Yokohama 226-8503, Japan b Biofunctional Catalyst Research Team, RIKEN Center for Sustainable Resource Science, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan c Chemical Genomics Research Group, RIKEN Center for Sustainable Resource Science, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan d Department of Chemistry and Materials Science, Graduate School of Science and Engineering, Tokyo Institute of Technology, 4259 Midori, Nagatsuta, Yokohama 226-8503, Japan * Corresponding author: [email protected] ABSTRACT The present study investigated extracellular electron transfer (EET) of Pseudomonas stutzeri, a model organism for bacterial denitrification. Electrochemical cultivation of P. stutzeri in a lithotrophic medium with Fe2+ ions generated the clear cathodic current associated with denitrification reactions. The EET ability of P. stutzeri was greatly strengthened by the addition of acetate, which correlated with the enhanced rate of the enzymatic oxidation of Fe2+. Since the addition of acetate induced the change of cellular metabolisms from lithotrophic denitrification to mixotrophic one, the present finding suggests the effectiveness of EET monitoring as a descriptor of bacterial denitrification activity and changing metabolisms according to the environmental conditions.
    [Show full text]
  • Pseudomonas Stutzeri As an Alternative Host for Membrane Proteins Manuel Sommer, Hao Xie* and Hartmut Michel*
    Sommer et al. Microb Cell Fact (2017) 16:157 DOI 10.1186/s12934-017-0771-0 Microbial Cell Factories RESEARCH Open Access Pseudomonas stutzeri as an alternative host for membrane proteins Manuel Sommer, Hao Xie* and Hartmut Michel* Abstract Background: Studies on membrane proteins are often hampered by insufcient yields of the protein of interest. Several prokaryotic hosts have been tested for their applicability as production platform but still Escherichia coli by far is the one most commonly used. Nevertheless, it has been demonstrated that in some cases hosts other than E. coli are more appropriate for certain target proteins. Results: Here we have developed an expression system for the heterologous production of membrane proteins using a single plasmid-based approach. The gammaproteobacterium Pseudomonas stutzeri was employed as a new production host. We investigated several basic microbiological features crucial for its handling in the laboratory. The organism belonging to bio-safety level one is a close relative of the human pathogen Pseudomonas aeruginosa. Pseu- domonas stutzeri is comparable to E. coli regarding its growth and cultivation conditions. Several efective antibiotics were identifed and a protocol for plasmid transformation was established. We present a workfow including cloning of the target proteins, small-scale screening for the best production conditions and fnally large-scale production in the milligram range. The GFP folding assay was used for the rapid analysis of protein folding states. In summary, out of 36 heterologous target proteins, 20 were produced at high yields. Additionally, eight transporters derived from P. aeruginosa could be obtained with high yields. Upscaling of protein production and purifcation of a Gluconate:H+ Symporter (GntP) family transporter (STM2913) from Salmonella enterica to high purity was demonstrated.
    [Show full text]
  • Analysis of Susceptibility Profile of Pseudomonas Spp. and Prevalence
    Revista de Ciências Farmacêuticas Básica e Aplicada Rev. Ciênc. Farm. Básica Apl., v. 26, n.2, p. 145-148, 2005 Journal of Basic and Applied Pharmaceutical Sciences ISSN 1808-4532 Analysis of susceptibility profile of Pseudomonas spp. and prevalence of bacterial samples from the surfaces of dental consulting-rooms Pietro, R.C.L.R.1,2*; Kashima, S.1; Almeida, A.M.F.1; Silva, C.H.P.M.3; Rocha, L.B.4; Pádua, J.M.4; Lia, R.C.C.4 1Curso de Ciências Farmacêuticas da Universidade de Ribeirão Preto,UNAERP; 2Departamento de Fármacos e Medicamentos da 3 4 Faculdade de Ciências Farmacêuticas de Araraquara, UNESP; Curso de Odontologia, FAESA; Curso de Odontologia da Universidade de Ribeirão Preto, UNAERP Recebido 15/09/05 / Aceito 21/11/05 ABSTRACT presence in the environment. The objective of this study was to analyze the The aim of this research was to evaluate the susceptibility susceptibility profile of P.aeruginosa, one of the most profile of Pseudomonas spp. and the prevalence of prevalent bacteria involved in cross-contamination, and the bacterial samples isolated from horizontal surfaces incidence of microbial contamination on surfaces in dental surrounding wash-basins used by dentists in several consulting-rooms, near to and distant from the dentists’ adjoined consulting-rooms, at points next to and at a wash basins, with the purpose of revealing any high risks distance from the basin, before and after surgical of infections that could be reduced with effective procedures. Our results showed a high percentage of contamination control procedures. Gram-positive cocci and Gram-negative bacilli; 34.66% were Staphylococcus spp.
    [Show full text]
  • Pseudomonas Stutzeri Prosthetic Valve Endocarditis: a Case Report And
    Journal of Infection and Public Health 12 (2019) 434–437 Contents lists available at ScienceDirect Journal of Infection and Public Health j ournal homepage: http://www.elsevier.com/locate/jiph Pseudomonas stutzeri prosthetic valve endocarditis: A case report and review of the literature a,1 b,1 b b,∗ Zeina Halabi , Michele Mocadie , Saeed El Zein , Souha S. Kanj a Department of Family Medicine, American University of Beirut Medical Center, Beirut, Lebanon b Department of Internal Medicine, Division of Infectious Diseases, American University of Beirut Medical Center, Beirut, Lebanon a r a t i b s c t l e i n f o r a c t Article history: We report a case of Pseudomonas stutzeri endocarditis in Lebanon. The patient had a recent history of Received 17 April 2018 prosthetic aortic valve replacement and presented to the emergency department with fever and chills. Received in revised form 1 July 2018 Transesophageal echocardiography confirmed the presence of a vegetation on the prosthetic valve and Accepted 5 July 2018 blood cultures yielded P. stutzeri. The patient was treated with surgery and antibiotics but deterio- rated and passed away four days after admission. To our knowledge, this is the fifth case of P. stutzeri Keywords: endocarditis reported in the literature, and the first case with early presentation and mortality. Pseudomonas stutzeri © 2018 The Authors. Published by Elsevier Limited on behalf of King Saud Bin Abdulaziz University Prosthetic valve Endocarditis for Health Sciences. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).
    [Show full text]
  • Phosphorus Metabolism in Pseudomonas Stutzeri
    AN ABSTRACT OF THE THESIS OF Lloyd Floren Elliott for the Ph. D. in Microbiology (Name) (Degree) (Major) Date thesis is presented ,L4.74. Title PHOSPHORUS METABOLISM IN PSEUDOMONAS STUTZERI Abstract approved ( or Professor) The role of nitrate oxygen as a terminal acceptor of hydrogen stands as a unique form of bacterial respiration. As nitrate is re- duced to nitrite on to the gaseous state (N2, N2O) the substrate is oxidized to give the requisite energy for cell growth. The present study will deal with selected aspects of the energy levels gained when nitrate and molecular oxygen operate as the terminal electron accep- tors. Pseudomonas stutzeri, an active nitrate reducer, was used throughout the entire study. The culture was maintained on nitrate agar. A semi - synthetic medium was used for the experimental studies. An electrolytic respirometer assembly was used for oxygen uptake and nitrogen evolution was measured using a Beckman model GC -2 chromatograph. Analyses for nitrites, total nitrogen, carbon dioxide and phosphorus were determined by established chemical procedures. Several older aspects of the problem have been clarified and new information presented. As might have been expected, large cell masses did not show a measurable uptake of phosphorus pre- sumably because of existing cell reserves. On the other hand a growing cell system did fix significant amounts of phosphorus and when oxygen uptake was measured, applicable growth PIO ratios were obtained. The aerobic system showed a relative PIO ratio of 0. 01 whereas the ratio for the anaerobic nitrate cell system was 0. 0036. The observed difference in the two ratios of 2.
    [Show full text]