Use of Genetically Engineered Microorganisms (GEMs) for the Bioremediation of Contaminants

ABSTRACT This paper presents a critical review of the literature on the application of genetically engineered microorganisms (GEMs) in bioremediation. The important aspects of using GEMs in bioremediation, such as development of novel strains with desirable properties through pathway construction and the modification of enzyme specificity and affinity, are discussed in detail. Particular attention is given to the genetic engineering of bacteria using bacterial hemoglobin (VHb) for the treatment of aromatic organic compounds under hypoxic conditions. The application of VHb technology may advance treatment of contaminated sites, where oxygen availability limits the growth of aerobic bioremediating bacteria, as well as the functioning of oxygenases required for mineralization of many organic pollutants. Despite the many advantages of GEMs, there are still concerns that their introduction into polluted sites to enhance bioremediation may have adverse environmental effects, such as gene transfer. The extent of horizontal gene transfer from GEMs in the environment, compared to that of native organisms including benefits regarding bacterial bioremediation that may occur as a result of such transfer, is discussed. Recent advances in tracking methods and containment strategies for GEMs, including several biological systems that have been developed to detect the fate of GEMs in the environment, are also summarized in this review. Critical research questions pertaining to the development and implementation of GEMs for enhanced bioremediation have been identified and posed for possible future research.

[1]  J. Bailey,et al.  Intracellular Expression of VitreoscillaHemoglobin (VHb) Enhances Total Protein Secretion and Improves the Production of α‐Amylase and Neutral Protease in Bacillus subtilis , 1996, Biotechnology progress.

[2]  Thomas Macek,et al.  Polychlorinated Biphenyl Rhizoremediation by Pseudomonas fluorescens F113 Derivatives, Using a Sinorhizobium meliloti nod System To Drive bph Gene Expression , 2005, Applied and Environmental Microbiology.

[3]  W. Stemmer,et al.  Molecular evolution of an arsenate detoxification pathway by DNA shuffling , 1997, Nature Biotechnology.

[4]  W. Suen,et al.  Cloning and characterization of Pseudomonas sp. strain DNT genes for 2,4-dinitrotoluene degradation , 1993, Journal of bacteriology.

[5]  F. Mondello,et al.  Identification and modification of biphenyl dioxygenase sequences that determine the specificity of polychlorinated biphenyl degradation , 1997, Applied and environmental microbiology.

[6]  S. Farrand,et al.  A novel gene tag for identifying microorganisms released into the environment , 1994, Applied and environmental microbiology.

[7]  H. Cha,et al.  Enhanced detoxification of organophosphates using recombinant Escherichia coli with co-expression of organophosphorus hydrolase and bacterial hemoglobin , 2002, Biotechnology Letters.

[8]  W. Reineke.  Development of hybrid strains for the mineralization of chloroaromatics by patchwork assembly. , 1998, Annual review of microbiology.

[9]  Z. Kawabata,et al.  Fate of a PCBS degrading recombinant pseudomonas putida AC30(PMFB2) and its effect on the densities of microbes in marine microcosms contaminated with PCBS , 1998 .

[10]  Plasmid transfer for enhancing degradation capabilities. , 1995 .

[11]  D. Webster,et al.  Genetic engineering to contain the Vitreoscilla hemoglobin gene enhances degradation of benzoic acid by Xanthomonas maltophilia. , 2000, Biotechnology and bioengineering.

[12]  A Mulchandani,et al.  Engineering of improved microbes and enzymes for bioremediation. , 1999, Current opinion in biotechnology.

[13]  H. Suenaga,et al.  Biphenyl Dioxygenases: Functional Versatilities and Directed Evolution , 2004, Journal of bacteriology.

[14]  Lisa Alvarez-Cohen,et al.  Aerobic biodegradation of N-nitrosodimethylamine (NDMA) by axenic bacterial strains. , 2005, Biotechnology and bioengineering.

[15]  B. Ensley,et al.  Biochemical diversity of trichloroethylene metabolism. , 1991, Annual review of microbiology.

[16]  Dirk Springael,et al.  Horizontal gene transfer and microbial adaptation to xenobiotics: new types of mobile genetic elements and lessons from ecological studies. , 2004, Trends in microbiology.

[17]  J. Martín,et al.  The Production of Cephalosporin C by Acremonium chrysogenum is Improved by the Intracellular Expression of a Bacterial Hemoglobin , 1993, Bio/Technology.

[18]  P. Riley,et al.  Value-enhanced crops: biotechnology's next stage , 1999 .

[19]  J. D. Díaz Ricci,et al.  Plasmid Effects on Escherichia coli Metabolism , 2000, Critical reviews in biotechnology.

[20]  D. Stahl,et al.  The role of genes in biological processes. Part 1 , 1990 .

[21]  J. Jansson,et al.  Quantification of the presence and activity of specific microorganisms in nature , 1997, Molecular biotechnology.

[22]  D. Stahl,et al.  Plasmid transfer for enhancing degradation capabilities. , 1995, Environmental health perspectives.

[23]  K. Dikshit,et al.  Oxygen dependent regulation of Vitreoscilla globin gene: evidence for positive regulation by FNR. , 1994, Biochemical and biophysical research communications.

[24]  G. Toranzos,et al.  Stability of manipulated plasmid DNA in aquatic environments , 1996 .

[25]  K. Timmis,et al.  Bacteria designed for bioremediation. , 1999, Trends in biotechnology.

[26]  Zhipei Liu,et al.  Novel Partial Reductive Pathway for 4-Chloronitrobenzene and Nitrobenzene Degradation in Comamonas sp. Strain CNB-1 , 2006, Applied and Environmental Microbiology.

[27]  J. Ramos,et al.  Characterization of Cell Lysis in Pseudomonas putida Induced upon Expression of Heterologous Killing Genes , 1998, Applied and Environmental Microbiology.

[28]  V. de Lorenzo,et al.  Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria , 1990, Journal of bacteriology.

[29]  Jonathan J Dennis,et al.  The evolution of IncP catabolic plasmids. , 2005, Current opinion in biotechnology.

[30]  F. Brockman.  Nucleic‐acid‐based methods for monitoring the performance of in situ bioremediation , 1995 .

[31]  Wilfred Chen,et al.  Detoxification of organophosphate nerve agents by immobilized dual functional biocatalysts in a cellulose hollow fiber bioreactor. , 2005, Biotechnology and bioengineering.

[32]  J. R. van der Meer,et al.  Low-Frequency Horizontal Transfer of an Element Containing the Chlorocatechol Degradation Genes fromPseudomonas sp. Strain B13 to Pseudomonas putidaF1 and to Indigenous Bacteria in Laboratory-Scale Activated-Sludge Microcosms , 1998, Applied and Environmental Microbiology.

[33]  D. Webster,et al.  The bacterial hemoglobin from Vitreoscilla can support the aerobic growth of Escherichia coli lacking terminal oxidases. , 1992, Archives of biochemistry and biophysics.

[34]  J. Tiedje,et al.  Quantitative Detection of Microbial Genes by Using DNA Microarrays , 2002, Applied and Environmental Microbiology.

[35]  M. Moran,et al.  Adaptation of model genetically engineered microorganisms to lake water: growth rate enhancements and plasmid loss , 1992, Applied and environmental microbiology.

[36]  M. Daly,et al.  Hg(II) sequestration and protection by the MerR metal-binding domain (MBD). , 2006, Microbiology.

[37]  S. Lindow,et al.  Relative Expression and Stability of a Chromosomally Integrated and Plasmid-Borne Marker Gene Fusion in Environmentally Competent Bacteria , 1997, Current Microbiology.

[38]  D. Webster,et al.  Chromosomal integration of the Vitreoscilla hemoglobin gene in Burkholderia and Pseudomonas for the purpose of producing stable engineered strains with enhanced bioremediating ability , 2001, Journal of Industrial Microbiology and Biotechnology.

[39]  V. de Lorenzo,et al.  Designing recombinant Pseudomonas strains to enhance biodesulfurization , 1997, Journal of bacteriology.

[40]  P. de Vos,et al.  Effect of Dissemination of 2,4-Dichlorophenoxyacetic Acid (2,4-D) Degradation Plasmids on 2,4-D Degradation and on Bacterial Community Structure in Two Different Soil Horizons , 2000, Applied and Environmental Microbiology.

[41]  R. Seidler,et al.  Effects of 2,4-Dichlorophenol, a Metabolite of a Genetically Engineered Bacterium, and 2,4-Dichlorophenoxyacetate on Some Microorganism-Mediated Ecological Processes in Soil , 1991, Applied and environmental microbiology.

[42]  M. R. Bruins,et al.  Pseudomonas pickettii: a common soil and groundwater aerobic bacteria with pathogenic and biodegradation properties. , 2000, Ecotoxicology and environmental safety.

[43]  W. V. van Zyl,et al.  The metabolic burden of the PGK1 and ADH2 promoter systems for heterologous xylanase production by Saccharomyces cerevisiae in defined medium. , 2001, Biotechnology and bioengineering.

[44]  A. Ohta,et al.  Novel Approach to the Improvement of Biphenyl and Polychlorinated Biphenyl Degradation Activity: Promoter Implantation by Homologous Recombination , 2003, Applied and Environmental Microbiology.

[45]  L. Bülow,et al.  Gene expression profiling of Escherichia coli expressing double Vitreoscilla haemoglobin. , 2004, Journal of biotechnology.

[46]  R. Seidler,et al.  Survival and degradative capacity of Pseudomonas putida induced or constitutively expressing plasmid-mediated degradation of 2,4-dichlorophenoxyacetate (TFD) in soil , 1990 .

[47]  L. Zentilin,et al.  A novel procedure for quantitative polymerase chain reaction by coamplification of competitive templates. , 1992, Gene.

[48]  S. Molin,et al.  Effects of genes exerting growth inhibition and plasmid stability on plasmid maintenance , 1987, Journal of bacteriology.

[49]  J. Ramos,et al.  Construction and behavior of biologically contained bacteria for environmental applications in bioremediation , 1995, Applied and environmental microbiology.

[50]  J. Bailey,et al.  Effect of Vitreoscilla hemoglobin dosage on microaerobic Escherichia coli carbon and energy metabolism , 1996, Biotechnology and bioengineering.

[51]  D. Webster,et al.  ArcA works with Fnr as a positive regulator of Vitreoscilla (bacterial) hemoglobin gene expression in Escherichia coli. , 2005, Microbiological research.

[52]  J. Pandey,et al.  Accessing microbial diversity for bioremediation and environmental restoration. , 2005, Trends in biotechnology.

[53]  L. Forney,et al.  Enhancement of 2,4-dichlorophenoxyacetic acid (2,4-D) degradation in soil by dissemination of catabolic plasmids , 2004, Antonie van Leeuwenhoek.

[54]  K. Timmis,et al.  Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. , 1981, Gene.

[55]  K. Furukawa.  Biochemical and genetic bases of microbial degradation of polychlorinated biphenyls (PCBs). , 2000, The Journal of general and applied microbiology.

[56]  A. Mulchandani,et al.  Detoxification of organophosphate nerve agents by immobilized Escherichia coli with surface-expressed organophosphorus hydrolase. , 1999, Biotechnology and bioengineering.

[57]  M. Stefanidou,et al.  Alternative Luciferase for Monitoring Bacterial Cells under Adverse Conditions , 2005, Applied and Environmental Microbiology.

[58]  H. Wand,et al.  Acquisition of a deliberately introduced phenol degradation operon, pheBA, by different indigenous Pseudomonas species , 1997, Applied and environmental microbiology.

[59]  S. Daunert,et al.  Novel reporter gene in a fluorescent‐based whole cell sensing system , 2006, Biotechnology and bioengineering.

[60]  D. Webster,et al.  Metabolic engineering of Serratia marcescens with the bacterial hemoglobin gene: alterations in fermentation pathways. , 1998, Biotechnology and bioengineering.

[61]  X. Nesme,et al.  Detection and enumeration of soil bacteria using the MPN-PCR technique , 1996 .

[62]  J. Ramos,et al.  Retrotransfer of DNA in the rhizosphere. , 2000, Environmental microbiology.

[63]  K. Mortelmans,et al.  Biodegradation of 2,4-dinitrotoluene by a Pseudomonas sp , 1991, Applied and environmental microbiology.

[64]  J. Tiedje,et al.  Evidence for Interspecies Gene Transfer in the Evolution of 2,4-Dichlorophenoxyacetic Acid Degraders , 1998, Applied and Environmental Microbiology.

[65]  Webster Da.  Structure and function of bacterial hemoglobin and related proteins. , 1988 .

[66]  T. Omori,et al.  Quantification of the carbazole 1,9a-dioxygenase gene by real-time competitive PCR combined with co-extraction of internal standards. , 2001, FEMS microbiology letters.

[67]  K. Timmis,et al.  An upstream XylR‐ and IHF‐induced nucleoprotein complex regulates the sigma 54‐dependent Pu promoter of TOL plasmid. , 1991, The EMBO journal.

[68]  D. Arp,et al.  Molecular and cellular fundamentals of aerobic cometabolism of trichloroethylene , 2004, Biodegradation.

[69]  J. Bailey,et al.  Fnr, a Global Transcriptional Regulator of Escherichia coli, Activates the Vitreoscilla Hemoglobin (VHb) Promoter and Intracellular VHb Expression Increases Cytochrome d Promoter Activity , 1995, Biotechnology progress.

[70]  W. Weigand,et al.  Effects of recombinant plasmid size on cellular processes in Escherichia coli. , 1987, Plasmid.

[71]  G. Fabro,et al.  Engineering Pseudomonas fluorescens for Biodegradation of 2,4-Dinitrotoluene , 2005, Applied and Environmental Microbiology.

[72]  D. Stahl,et al.  The specific growth rate of Pseudomonas putida PAW1 influences the conjugal transfer rate of the TOL plasmid , 1993, Applied and environmental microbiology.

[73]  D. Webster,et al.  Enhancement of 2,4-dinitrotoluene biodegradation by Burkholderia sp. in sand bioreactors using bacterial hemoglobin technology , 2004, Biodegradation.

[74]  J. Gerritse,et al.  Isolation of Alcaligenes sp. strain L6 at low oxygen concentrations and degradation of 3-chlorobenzoate via a pathway not involving (chloro)catechols , 1996, Applied and environmental microbiology.

[75]  J. Ramos,et al.  Suicidal genetic elements and their use in biological containment of bacteria. , 1993, Annual review of microbiology.

[76]  D. Webster.  Structure and function of bacterial hemoglobin and related proteins. , 1988, Advances in inorganic biochemistry.

[77]  B. Jank,et al.  Assessing the environmental impacts of transgenic plants. , 2001, Trends in biotechnology.

[78]  Z. Kawabata,et al.  Factors affecting the survival of genetically engineered Escherichia coli bearing a plasmid in a paddy field microcosm , 1998 .

[79]  J. Davison,et al.  Genetic exchange between bacteria in the environment. , 1999, Plasmid.

[80]  D. Webster,et al.  Genetic engineering of Serratia marcescens with bacterial hemoglobin gene: effects on growth, oxygen utilization, and cell size. , 1998, Biotechnology and bioengineering.

[81]  D. Paul,et al.  Conceptualizing "suicidal genetically engineered microorganisms" for bioremediation applications. , 2005, Biochemical and biophysical research communications.

[82]  F. Arnold,et al.  Functional Expression of a Fungal Laccase in Saccharomyces cerevisiae by Directed Evolution , 2003, Applied and Environmental Microbiology.

[83]  J. Tommassen,et al.  Construction of phoE-caa, a novel PCR- and immunologically detectable marker gene for Pseudomonas putida , 1994, Applied and environmental microbiology.

[84]  J. Bailey,et al.  Actinorhodin Production By Streptomyces coelicolor and Growth of Streptomyces lividans Are Improved by the Expression of a Bacterial Hemoglobin , 1991, Bio/Technology.

[85]  J. R. van der Meer,et al.  Genomic islands and the evolution of catabolic pathways in bacteria. , 2003, Current opinion in biotechnology.

[86]  A. Howard,et al.  Vitreoscilla Hemoglobin Binds to Subunit I of Cytochrome bo Ubiquinol Oxidases* , 2002, The Journal of Biological Chemistry.

[87]  G. Fox,et al.  A prototype stable RNA identification cassette for monitoring plasmids of genetically engineered microorganisms. , 1993, Systematic and applied microbiology.

[88]  C R Cantor,et al.  A new approach for containment of microorganisms: dual control of streptavidin expression by antisense RNA and the T7 transcription system. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[89]  K. Pagilla,et al.  Enhanced kinetics of genetically engineered Burkholderia cepacia: the role of vgb in the hypoxic metabolism of 2‐CBA , 2004, Biotechnology and bioengineering.

[90]  D. Webster,et al.  Effects of Vitreoscilla hemoglobin on the 2,4-dinitrotoluene (2,4-DNT) dioxygenase activity of Burkholderia and on 2,4-DNT degradation in two-phase bioreactors , 2003, Journal of Industrial Microbiology and Biotechnology.

[91]  F. Brockman,et al.  Direct Detection of 16S rRNA in Soil Extracts by Using Oligonucleotide Microarrays , 2001, Applied and Environmental Microbiology.

[92]  D. Webster,et al.  Cloning and Expression of Vitreoscilla Hemoglobin Gene in Burkholderia sp. Strain DNT for Enhancement of 2,4‐Dinitrotoluene Degradation , 2000, Biotechnology progress.

[93]  I. Pepper,et al.  Gene transfer of Alcaligenes eutrophus JMP134 plasmid pJP4 to indigenous soil recipients , 1996, Applied and environmental microbiology.

[94]  M. Moran,et al.  Impact of a genetically engineered bacterium with enhanced alkaline phosphatase activity on marine phytoplankton communities , 1996, Applied and environmental microbiology.

[95]  D. Webster,et al.  Cloning and expression of the Vitreoscilla hemoglobin gene in pseudomonads: effects on cell growth , 1995, Applied Microbiology and Biotechnology.

[96]  S T E V E N R I P P,et al.  Controlled Field Release of a Bioluminescent Genetically Engineered Microorganism for Bioremediation Process Monitoring and Control , 2000 .

[97]  L. Wackett,et al.  Field-scale remediation of atrazine-contaminated soil using recombinant Escherichia coli expressing atrazine chlorohydrolase. , 2000, Environmental microbiology.

[98]  C. Rensing,et al.  New Approaches for Bioaugmentation as a Remediation Technology , 2004 .

[99]  Dorothea K. Thompson,et al.  Development and Evaluation of Functional Gene Arrays for Detection of Selected Genes in the Environment , 2001, Applied and Environmental Microbiology.

[100]  Wilfred Chen,et al.  Specific Adhesion to Cellulose and Hydrolysis of Organophosphate Nerve Agents by a Genetically Engineered Escherichia coli Strain with a Surface-Expressed Cellulose-Binding Domain and Organophosphorus Hydrolase , 2002, Applied and Environmental Microbiology.

[101]  J. Jansson.  Tracking genetically engineered microorganisms in nature. , 1995, Current opinion in biotechnology.

[102]  J. Keasling,et al.  Uranyl Precipitation by Pseudomonas aeruginosa via Controlled Polyphosphate Metabolism , 2004, Applied and Environmental Microbiology.

[103]  K. Furukawa.  Engineering dioxygenases for efficient degradation of environmental pollutants. , 2000, Current opinion in biotechnology.

[104]  D. Chandler,et al.  Redefining relativity: quantitative PCR at low template concentrations for industrial and environmental microbiology , 1998, Journal of Industrial Microbiology and Biotechnology.

[105]  D. Webster,et al.  Structure-function studies of the Vitreoscilla hemoglobin D-region. , 2004, Biochemical and biophysical research communications.

[106]  W. Deckwer,et al.  Plasmid stability of recombinant Pseudomonas sp. B13 FR1 pFRC20P in continuous culture. , 1998, Biotechnology and bioengineering.

[107]  K. Pagilla,et al.  Comparison of 2-chlorobenzoic acid biodegradation in a membrane bioreactor by B. cepacia and B. cepacia bearing the bacterial hemoglobin gene. , 2006, Water research.

[108]  Juan L. Ramos,et al.  Construction of an Efficient Biologically ContainedPseudomonas putida Strain and Its Survival in Outdoor Assays , 1998, Applied and Environmental Microbiology.

[109]  J. Bailey,et al.  Intracellular expression of Vitreoscilla hemoglobin alters Escherichia coli energy metabolism under oxygen-limited conditions. , 1994, European journal of biochemistry.

[110]  V. de Lorenzo,et al.  Mini‐transposons in microbial ecology and environmental biotechnology , 1998, FEMS Microbiology Ecology.

[111]  D. Webster,et al.  Improvement of bioremediation by Pseudomonas and Burkholderia by mutants of the Vitreoscilla hemoglobin gene (vgb) integrated into their chromosomes , 2005, Journal of Industrial Microbiology and Biotechnology.

[112]  D. Newby,et al.  Comparison of 2,4-Dichlorophenoxyacetic Acid Degradation and Plasmid Transfer in Soil Resulting from Bioaugmentation with Two Different pJP4 Donors , 2000, Applied and Environmental Microbiology.

[113]  J. Quensen,et al.  Construction and Characterization of Two Recombinant Bacteria That Grow on ortho- and para-Substituted Chlorobiphenyls , 1999, Applied and Environmental Microbiology.

[114]  R. Sharp,et al.  Activity and stability of a recombinant plasmid-borne TCE degradative pathway in suspended cultures. , 1998, Biotechnology and bioengineering.

[115]  C. Collins,et al.  Safe biotechnology 9: values in risk assessment for the environmental application of microorganisms. The Safety in Biotechnology Working Party of the European Federation of Biotechnology. , 1999, Trends in biotechnology.

[116]  Sven Panke,et al.  Engineering of Quasi-Natural Pseudomonas putida Strains for Toluene Metabolism through anortho-Cleavage Degradation Pathway , 1998, Applied and Environmental Microbiology.

[117]  N. Boon,et al.  Catabolic mobile genetic elements and their potential use in bioaugmentation of polluted soils and waters. , 2002, FEMS microbiology ecology.

[118]  V. V. Velkov,et al.  Stress-induced evolution and the biosafety of genetically modified microorganisms released into the environment , 2001, Journal of Biosciences.

[119]  D. Gibson,et al.  Aromatic hydrocarbon dioxygenases in environmental biotechnology. , 2000, Current opinion in biotechnology.

[120]  D. Webster,et al.  Sequence of the region downstream of theVitreoscilla hemoglobin gene:vgb is not part of a multigene operon , 1994, Applied Microbiology and Biotechnology.

[121]  L. Wackett,et al.  The Atrazine Catabolism Genes atzABC Are Widespread and Highly Conserved , 1998, Journal of bacteriology.

[122]  R. Gupta,et al.  Effect of growth environment on the stability of a recombinant shuttle plasmid, pCPPS-31, in Escherichia coli. , 1995, Journal of biotechnology.

[123]  J. Cairns,et al.  Genetically Engineered Erwinia carotovora in Aquatic Microcosms: Survival and Effects on Functional Groups of Indigenous Bacteria , 1989, Applied and Environmental Microbiology.

[124]  S. Lindow.  The use of reporter genes in the study of microbial ecology , 1995 .

[125]  A. Schmid,et al.  Biodegradation, Biotransformation, and Biocatalysis (B3) , 2002, Applied and Environmental Microbiology.

[126]  M. Mergeay,et al.  Community shifts in a seeded 3-chlorobenzoate degrading membrane biofilm reactor: indications for involvement of in situ horizontal transfer of the clc-element from inoculum to contaminant bacteria. , 2002, Environmental microbiology.

[127]  M. Suidan,et al.  Challenges in Biodegradation of Trace Organic Contaminants—Gasoline Oxygenates and Sex Hormones , 2005, Water environment research : a research publication of the Water Environment Federation.

[128]  A. Singh,et al.  Composting: A Potentially Safe Process for Disposal of Genetically Modified Organisms , 2006, Critical reviews in biotechnology.

[129]  J. Keasling,et al.  Dibenzothiophene biodesulfurization pathway improvement using diagnostic GFP fusions , 2004, Biotechnology and Bioengineering.

[130]  Investigation of the effect of growth environment on the stability of low-copy-number plasmids in Escherichia coli. , 1987, Journal of general microbiology.

[131]  T. Omori,et al.  Recent developments in molecular techniques for identification and monitoring of xenobiotic-degrading bacteria and their catabolic genes in bioremediation , 2002, Applied Microbiology and Biotechnology.

[132]  Robert K. Colwell,et al.  The Planned Introduction of Genetically Engineered Organisms: Ecological Considerations and Recommendations , 1989 .

[133]  J E Bailey,et al.  Plasmid presence changes the relative levels of many host cell proteins and ribosome components in recombinant Escherichia coli , 1991, Biotechnology and bioengineering.

[134]  Hyung Joon Cha,et al.  Enhanced Biodegradation of Toxic Organophosphate Compounds Using Recombinant Escherichia coli with Sec Pathway‐Driven Periplasmic Secretion of Organophosphorus Hydrolase , 2006, Biotechnology progress.

[135]  Ashok Mulchandani,et al.  Biodegradation of organophosphorus pesticides by surface-expressed organophosphorus hydrolase , 1997, Nature Biotechnology.

[136]  A. Schmid,et al.  Recombinant Chlorobenzene Dioxygenase from Pseudomonas sp. P51: A Biocatalyst for Regioselective Oxidation of Aromatic Nitriles , 2005 .

[137]  G. Grundmann,et al.  Impact of the Microscale Distribution of a Pseudomonas Strain Introduced into Soil on Potential Contacts with Indigenous Bacteria , 2005, Applied and Environmental Microbiology.

[138]  J. R. van der Meer,et al.  Evolution of a chlorobenzene degradative pathway among bacteria in a contaminated groundwater mediated by a genomic island in Ralstonia. , 2003, Environmental Microbiology.

[139]  Gary S. Sayler,et al.  Induction of the tod Operon by Trichloroethylene in Pseudomonas putida TVA8 , 1998, Applied and Environmental Microbiology.

[140]  Jeffrey Philip Obbard,et al.  Recent advances in the bioremediation of persistent organic pollutants via biomolecular engineering , 2005 .

[141]  D. Paul,et al.  Suicidal genetically engineered microorganisms for bioremediation: need and perspectives. , 2005, BioEssays : news and reviews in molecular, cellular and developmental biology.

[142]  Lawrence P. Wackett,et al.  Engineering a recombinant Deinococcus radiodurans for organopollutant degradation in radioactive mixed waste environments , 1998, Bio/Technology.

[143]  D. Webster,et al.  Biodegradation of 2-Chlorobenzoate by Recombinant Burkholderia Cepacia Expressing Vitreoscilla Hemoglobin Under Variable Levels of Oxygen Availability , 2003, Biodegradation.

[144]  R. Fulthorpe,et al.  Involvement of a chlorobenzoate-catabolic transposon, Tn5271, in community adaptation to chlorobiphenyl, chloroaniline, and 2,4-dichlorophenoxyacetic acid in a freshwater ecosystem , 1992, Applied and environmental microbiology.

[145]  T. Wood,et al.  Rhizoremediation of Trichloroethylene by a Recombinant, Root-Colonizing Pseudomonas fluorescensStrain Expressing Toluene ortho-Monooxygenase Constitutively , 1998, Applied and Environmental Microbiology.

[146]  Juan L. Ramos,et al.  Dual System To Reinforce Biological Containment of Recombinant Bacteria Designed for Rhizoremediation , 2001, Applied and Environmental Microbiology.

[147]  D. Webster,et al.  Effects of Culture Conditions on Enhancement of 2,4‐Dinitrotoluene Degradation by BurkholderiaEngineered with the Vitreoscilla Hemoglobin Gene , 2001, Biotechnology progress.

[148]  G S Sayler,et al.  Field applications of genetically engineered microorganisms for bioremediation processes. , 2000, Current opinion in biotechnology.

[149]  D. Newby,et al.  Detection and Characterization of Plasmid pJP4 Transfer to Indigenous Soil Bacteria , 2000, Applied and Environmental Microbiology.

[150]  C. Nakatsu,et al.  Evidence for Acquisition in Nature of a Chromosomal 2,4-Dichlorophenoxyacetic Acid/(alpha)-Ketoglutarate Dioxygenase Gene by Different Burkholderia spp , 1996, Applied and environmental microbiology.

[151]  J. Bailey,et al.  Effect of Vitreoscilla hemoglobin expression on growth and specific tissue plasminogen activator productivity in recombinant chinese hamster ovary cells. , 1994, Biotechnology and bioengineering.

[152]  D. Newby,et al.  The role of selective pressure and selfish DNA in horizontal gene transfer and soil microbial community adaptation , 2002 .

[153]  V. de Lorenzo,et al.  Mini-transposons in microbial ecology and environmental biotechnology , 1998, FEMS Microbiology Ecology.

[154]  A. Goldman,et al.  Substrate Specificity of and Product Formation by Muconate Cycloisomerases: an Analysis of Wild-Type Enzymes and Engineered Variants , 1998, Applied and Environmental Microbiology.

[155]  D. Webster,et al.  Presence of the bacterial hemoglobin gene improves α-amylase production of a recombinantEscherichia coli strain , 1990 .

[156]  Paul Keim,et al.  Development and Testing of a Bacterial Biosensor for Toluene-Based Environmental Contaminants , 1998, Applied and Environmental Microbiology.

[157]  J. Bailey,et al.  Transgenic tobacco expressing Vitreoscilla hemoglobin exhibits enhanced growth and altered metabolite production , 1997, Nature Biotechnology.

[158]  S. Molin,et al.  Application of a strain-specific rRNA oligonucleotide probe targeting Pseudomonas fluorescens Ag1 in a mesocosm study of bacterial release into the environment , 1995, Applied and environmental microbiology.

[159]  D. Webster,et al.  Vitreoscilla hemoglobin enhances the first step in 2,4-dinitrotoluene degradation in vitro and at low aeration in vivo , 2000 .

[160]  Ramandeep,et al.  Vitreoscilla Hemoglobin , 2001, The Journal of Biological Chemistry.

[161]  V. de Lorenzo,et al.  Resistance to Tellurite as a Selection Marker for Genetic Manipulations of Pseudomonas Strains , 1998, Applied and Environmental Microbiology.

[162]  L. Wong,et al.  Engineering molecular recognition in alkane oxidation catalysed by cytochrome P450cam , 1998 .