← Back to Search
Purification To Homogeneity And Characterization Of A Novel Pseudomonas Putida Chromate Reductase
C. H. Park, M. Keyhan, B. Wielinga, S. Fendorf, A. Matin
Published 2000 · Chemistry, Medicine
Download PDFAnalyze on Scholarcy
ABSTRACT Cr(VI) (chromate) is a widespread environmental contaminant. Bacterial chromate reductases can convert soluble and toxic chromate to the insoluble and less toxic Cr(III). Bioremediation can therefore be effective in removing chromate from the environment, especially if the bacterial propensity for such removal is enhanced by genetic and biochemical engineering. To clone the chromate reductase-encoding gene, we purified to homogeneity (>600-fold purification) and characterized a novel soluble chromate reductase from Pseudomonas putida, using ammonium sulfate precipitation (55 to 70%), anion-exchange chromatography (DEAE Sepharose CL-6B), chromatofocusing (Polybuffer exchanger 94), and gel filtration (Superose 12 HR 10/30). The enzyme activity was dependent on NADH or NADPH; the temperature and pH optima for chromate reduction were 80°C and 5, respectively; and theKm was 374 μM, with aVmax of 1.72 μmol/min/mg of protein. Sulfate inhibited the enzyme activity noncompetitively. The reductase activity remained virtually unaltered after 30 min of exposure to 50°C; even exposure to higher temperatures did not immediately inactivate the enzyme. X-ray absorption near-edge-structure spectra showed quantitative conversion of chromate to Cr(III) during the enzyme reaction.
This paper references
Use of starvation promoters to limit growth and selectively express trichloroethylene and phenol transformation activity in recombinant
A. Matin (1995)
Chemical contaminants on DOE lands and selection of contaminant mixtures for subsurface science research
R. G. Riley (1992)
SDS polyacrylamide gel electrophoresis of proteins.
B. Smith (1994)
RECOMBINANT DNA TECHNOLOGY II
R. Bajpai (1994)
Simplified method for silver staining of proteins in polyacrylamide gels and the mechanism of silver staining
J. Heukeshoven (1985)
Ground-Water Treatment for Chlorinated Solvents
P. Mccarty (1994)
A 13-element Ge detector for fluorescence EXAFS
Stephen P. Cramer (1988)
Standard Methods for the examination of water and wastewater
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
U. Laemmli (1970)
Molecular basis of hexavalent chromium carcinogenicity: effect on gene expression.
K. Wetterhahn (1989)
Mutagenicity of chromates in bacteria and its relevance to chromate carcinogenesis
S. Venitt (1974)
Use of Starvation Promoters To Limit Growth and Select for Trichloroethylene and Phenol Transformation Activity in Recombinant Escherichia coli
A. Matin (1995)
Chromium reduction in Pseudomonas putida.
Y. Ishibashi (1990)
PUTIDA CHROMATE REDUCTASE
Reduction of Hexavalent Chromium by Amorphous Iron Sulfide
Ronald R. Patterson and (1997)
Starvation Promoters of Escherichia coli: Their Function, Regulation, and Use in Bioprocessing and Bioremediation
A. Matin (1994)
Reduction of chromate by amorphous iron sulfide
R R Patterson (1997)
Designing microorganisms for the treatment of toxic wastes.
K. Timmis (1994)
In-situ bioremediation of soils and groundwater contaminated with petroleum hydrocarbons, p. 17–37
R. D. Norris (1994)
A carbon starvation survival gene of Pseudomonas putida is regulated by sigma 54.
Y. Kim (1995)
Isolation and Characterization of an Enterobacter cloacae Strain That Reduces Hexavalent Chromium under Anaerobic Conditions.
P. C. Wang (1989)
Use of starvation promoters to limit growth and selectively enrich expression of trichloroethylene- and phenol-transforming activity in recombinant Escherichia coli [corrected].
A. Matin (1995)
Decreased chromate uptake in Pseudomonas fluorescens carrying a chromate resistance plasmid.
H. Ohtake (1987)
In-situ bioremediation of soils and groundwater contaminated with petroleum hydrocarbons Handbook of bioremediation
R D Norris (1994)
NAD(P)H-dependent chromium (VI) reductase of Pseudomonas ambigua G-1: a Cr(V) intermediate is formed during the reduction of Cr(VI) to Cr(III).
T. Suzuki (1992)
The complete general secretory pathway in gram-negative bacteria.
A. Pugsley (1993)
Fate and Transport of Hexavalent Chromium in Undisturbed Heterogeneous Soil
P. M. Jardine (1999)
J Stohr (1992)
This paper is referenced by
Enzyme technology and biological remediation
C. Whiteley (2006)
Reduction of Hexavalent Chromium by Cell-Free Extract of Bacillus sphaericus AND 303 Isolated from Serpentine Soil
Arundhati Pal (2005)
Isolation of Cr(VI) reducing bacteria from industrial effluents and their potential use in bioremediation of chromium containing wastewater.
A. Zahoor (2009)
Complete genome sequence of Pseudomonas stutzeri strain RCH2 isolated from a Hexavalent Chromium [Cr(VI)] contaminated site
R. Chakraborty (2017)
Chromium(VI) Resistance and Extracellular Polysaccharide (EPS) Synthesis by Pseudomonas, Stenotrophomonas and Methylobacterium Strains
Sahlan Ozturk (2008)
Characterization of two genes involved in chromate resistance in a Cr(VI)-hyper-resistant bacterium
Francesca Decorosi (2009)
Simultaneous aerobic denitrification and Cr(VI) reduction by Pseudomonas brassicacearum LZ-4 in wastewater.
X. Yu (2016)
Cr(VI) reduction by cell-free extract of thermophillic Bacillus fusiformis NTR9
P. Pattanapipitpaisal (2013)
Occurrence and evaluation of chromium reducing bacteria in seepage water from chromite mine quarries of Orissa, India.
Satarupa Dey (2010)
The Role of Microbial Mats in the Removal of Hexavalent Chromium and Associated Shifts in Their Bacterial Community Composition
R. M. Abed (2020)
Optimization of Chromate Reduction by Whole Cells of Arthrobacter sp. SUK 1205 Isolated from Metalliferous Chromite Mine Environment
Satarupa Dey (2012)
Factors Affecting Detoxification of Hexavalent Chromium into Trivalent in Industrial Effluents by Indigenous Bacteria
Tasnim Ahmad (2015)
Purification of Chromate(VI) Reductase from Lichen photobiont, Trebouxia erici and its Effect on Wastewaters.
Efe M. Omwirhiren (2016)
Hexavalent chromium reduction by immobilized cells of Bacillus sphaericus AND 303
Arundhati Pal (2013)
Removal and reduction of chromium by Pseudomonas spp. and their correlation to rhamnolipid production.
Sahlan Ozturk (2012)
Enzyme improvement in the absence of structural knowledge: a novel statistical approach
Y. Barak (2008)
Bioremediation of chromium solutions and chromium containing wastewaters
P. Malaviya (2016)
Sorption and Transformation of Toxic Metals by Microorganisms
X. Han (2010)
Chromium reducing and plant growth promoting novel strain Pseudomonas aeruginosa OSG41 enhance chickpea growth in chromium amended soils
M. Oves (2013)
Mechanism of chromate reduction by the Escherichia coli protein, NfsA, and the role of different chromate reductases in minimizing oxidative stress during chromate reduction.
D. Ackerley (2004)
Surface-Enhanced Raman Imaging of Intracellular Bioreduction of Chromate in Shewanella oneidensis
S. Ravindranath (2011)
Isolation and Characterization of Heavy Metal Resistant Bacteria from Barak River Contaminated with Pulp Paper Mill Effluent
South Assam (2012)
Biochemical and molecular mechanisms involved in simultaneous phenol and Cr(VI) removal by Acinetobacter guillouiae SFC 500-1A
O. Ontañon (2015)
Influence of metal ions on biofilm formation by Arthrobacter sp. SUK 1205 and evaluation of their Cr(VI) removal efficacy
Satarupa Dey (2018)
Biomolecular Strategy To Decrease ChromateToxicity To Remediating Bacteria
D. Ackerley (2005)
Comparative genomics of 16 Microbacterium spp. that tolerate multiple heavy metals and antibiotics
D. R. Learman (2019)
Mechanisms of bacterial resistance to chromium compounds
M. I. Ramírez-Díaz (2007)
Cr (VI) Reduction by Indigenous Bacillus Species PB5 Isolated from Contaminated Soil of Abeokuta Ogun State, Nigeria
P. A. Wani (2015)
Prospective of chromium (VI) reduction under in vitro and in vivo conditions and stimulation of antioxidant defense of cowpea under the exposure of Cr (VI)
P. A. Wani (2018)
Characterization of a Cr(VI)-sensitive Pseudomonas corrugata 28 mutant impaired in a pyridine nucleotide transhydrogenase gene.
Francesca Decorosi (2011)
Isolation and Characterization of Heavy Metal Resistant Bacteria from Barak River Contaminated with Pulp Paper Mill Effluent, South Assam
Bibhas Rajkumar (2012)
Novel Biochemical Properties and Physiological Role of the Flavin Mononucleotide Oxidoreductase YhdA from Bacillus subtilis
Luz I Valenzuela-García (2020)See more