Online citations, reference lists, and bibliographies.
← Back to Search

Escherichia Coli NemA Is An Efficient Chromate Reductase That Can Be Biologically Immobilized To Provide A Cell Free System For Remediation Of Hexavalent Chromium

K. Robins, D. Hooks, B. H. Rehm, D. Ackerley
Published 2013 · Chemistry, Medicine

Cite This
Download PDF
Analyze on Scholarcy
Hexavalent chromium is a serious and widespread environmental pollutant. Although many bacteria have been identified that can transform highly water-soluble and toxic Cr(VI) to insoluble and relatively non-toxic Cr(III), bacterial bioremediation of Cr(VI) pollution is limited by a number of issues, in particular chromium toxicity to the remediating cells. To address this we sought to develop an immobilized enzymatic system for Cr(VI) remediation. To identify novel Cr(VI) reductase enzymes we first screened cell extracts from an Escherichia coli library of soluble oxidoreductases derived from a range of bacteria, but found that a number of these enzymes can reduce Cr(VI) indirectly, via redox intermediates present in the crude extracts. Instead, activity assays for 15 candidate enzymes purified as His6-tagged proteins identified E. coli NemA as a highly efficient Cr(VI) reductase (kcat/KM  = 1.1×105 M−1s−1 with NADH as cofactor). Fusion of nemA to the polyhydroxyalkanoate synthase gene phaC from Ralstonia eutropha enabled high-level biosynthesis of functionalized polyhydroxyalkanoate granules displaying stable and active NemA on their surface. When these granules were combined with either Bacillus subtilis glucose dehydrogenase or Candida boidinii formate dehydrogenase as a cofactor regenerating partner, high levels of chromate transformation were observed with only low initial concentrations of expensive NADH cofactor being required, the overall reaction being powered by consumption of the cheap sacrificial substrates glucose or formic acid, respectively. This system therefore offers promise as an economic solution for ex situ Cr(VI) remediation.
This paper references
In vivo monitoring of PHA granule formation using GFP-labeled PHA synthases.
V. Peters (2005)
uvrB gene deletion enhances SOS chromotest sensitivity for nitroreductases that preferentially generate the 4-hydroxylamine metabolite of the anti-cancer prodrug CB1954.
G.A. Prosser (2010)
A Soluble Flavoprotein Contributes to Chromate Reduction and Tolerance by Pseudomonas putida
A. (2003)
Future risk from a hypothesized RCRA site disposing of carcinogenic metals should a loss of societal memory occur
D. Okrent (1993)
Discovery and evaluation of Escherichia coli nitroreductases that activate the anti-cancer prodrug CB1954.
G. Prosser (2010)
Analysis of Novel Soluble Chromate and Uranyl Reductases and Generation of an Improved Enzyme by Directed Evolution
Y. Barak (2006)
In Vivo Enzyme Immobilization by Use of Engineered Polyhydroxyalkanoate Synthase
V. Peters (2006)
Mechanisms of bacterial resistance to chromium compounds
M. I. Ramírez-Díaz (2007)
Biotransformation of Explosives by the Old Yellow Enzyme Family of Flavoproteins
R. E. Williams (2004)
Ground-Water Treatment for Chlorinated Solvents
P. Mccarty (1994)
High cell-density culture of Escherichia coli.
S. Lee (1996)
Acceleration of azo dye decolorization by using quinone reductase activity of azoreductase and quinone redox mediator.
Guangfei Liu (2009)
Class I and III polyhydroxyalkanoate synthases from Ralstonia eutropha and Allochromatium vinosum: characterization and substrate specificity studies.
W. Yuan (2001)
Chemical contaminants on DOE lands and selection of contaminant mixtures for subsurface science research
R. G. Riley (1992)
Bacterial polymers: biosynthesis, modifications and applications
B. Rehm (2010)
Industrial biocatalysis today and tomorrow
A. Schmid (2001)
Glucose Dehydrogenase for the Regeneration of NADPH and NADH
Andrea Weckbecker (2005)
Molecular cloning of the nemA gene encoding N-ethylmaleimide reductase from Escherichia coli.
K. Miura (1997)
Vibrio harveyi Nitroreductase Is Also a Chromate Reductase
Young Hak Kwak (2003)
Discovery and evaluation of Escherichia coli nitroreductases that activate the anticancer prodrug CB1954
GA Prosser (2010)
Chromium in Drinking Water: Sources, Metabolism, and Cancer Risks
A. Zhitkovich (2011)
Bioremediation of Soils Polluted with Hexavalent Chromium using Bacteria: A Challenge
C. Viti (2007)
Effect of chromate stress on Escherichia coli K-12.
D. Ackerley (2006)
Immobilization of organophosphohydrolase OpdA from Agrobacterium radiobacter by overproduction at the surface of polyester inclusions inside engineered Escherichia coli
P. Blatchford (2012)
Cofactor regeneration for sustainable enzymatic biosynthesis.
Wenfang Liu (2007)
Creation and screening of a multi-family bacterial oxidoreductase library to discover novel nitroreductases that efficiently activate the bioreductive prodrugs CB1954 and PR-104A.
G. Prosser (2013)
Chromate-Reducing Properties of Soluble Flavoproteins from Pseudomonas putida and Escherichia coli
D. Ackerley (2004)
Replacement of the catalytic nucleophile cysteine-296 by serine in class II polyhydroxyalkanoate synthase from Pseudomonas aeruginosa-mediated synthesis of a new polyester: identification of catalytic residues.
A. A. Amara (2003)
Chromate reductase activity of the Paracoccusdenitrificans ferric reductase B (FerB) protein and its physiological relevance
Vojtěch Sedláček (2010)
Recent developments in pyridine nucleotide regeneration.
W. A. van der Donk (2003)
Remediation of Chromium and Uranium Contamination by Microbial Activity
Y. Cheng (2012)
High‐resolution structures of formate dehydrogenase from Candida boidinii
K. Schirwitz (2007)
Aqueous geochemistry of chromium: A review
F. Richard (1991)
Mutagenicity of inorganic compounds in Salmonella typhimurium: arsenic, chromium and selenium
G. Loefroth (1978)
Standard Methods for the examination of water and wastewater
Awwa (1999)
Tolerance of the Ralstonia eutropha Class I Polyhydroxyalkanoate Synthase for Translational Fusions to Its C Terminus Reveals a New Mode of Functional Display
A. C. Jahns (2009)
Multifunctional inorganic-binding beads self-assembled inside engineered bacteria.
A. C. Jahns (2008)
Molecular Dynamics of the Shewanella oneidensis Response to Chromate Stress*S
S. Brown (2006)
A novel chromate reductase from Thermus scotoductus SA-01 related to old yellow enzyme.
D. Opperman (2008)
Surface-mediated chromate-resistant mechanism of Enterobacter cloacae bacteria investigated by atomic force microscopy.
Chunpeng Yang (2007)
Reaction of chromium(VI) with ascorbate produces chromium(V), chromium(IV), and carbon-based radicals.
D. Stearns (1994)
Microbial community response to addition of polylactate compounds to stimulate hexavalent chromium reduction in groundwater.
Eoin L. Brodie (2011)
A membrane-associated protein with Cr(VI)-reducing activity from Thermus scotoductus SA-01.
D. Opperman (2008)
Kinetic and structural characterisation of Escherichia coli nitroreductase mutants showing improved efficacy for the prodrug substrate CB1954.
P. Race (2007)
In vivo reduction of chromium (VI) and its related free radical generation
K. J. Liu (2004)
Cr(VI) Remediation by PHA Bead immobilized NemA PLOS ONE | www.plosone
Handbook of Bioremediation
R. D. Norris (1993)

This paper is referenced by
Operational Characteristics of Immobilized Ochrobactrum sp. CUST210-1 Biosystem and Immobilized Chromate Reductase Biosystem in Continuously Treating Actual Chromium-Containing Wastewater
Gueyhorng Wang (2020)
Metatranscriptomic analysis reveals active microbes and genes responded to short-term Cr(VI) stress
Zhengsheng Yu (2020)
Bioengineering towards self-assembly of particulate vaccines.
B. H. Rehm (2017)
Chromate Interaction with the Chromate Reducing Actinobacterium Intrasporangium chromatireducens Q5-1
H. Liu (2015)
NfoR: Chromate Reductase or Flavin Mononucleotide Reductase?
Audrey G. O’Neill (2020)
Use of Bacterial Polyhydroxyalkanoates in Protein Display Technologies
I. Hay (2014)
Tibet plateau probiotic mitigates chromate toxicity in mice by alleviating oxidative stress in gut microbiota
Pengya Feng (2020)
Detoxification and Bioremediation of Hexavalent Chromium Using Microbes and Their Genes: An Insight into Genomic, Proteomic and Bioinformatics Studies
H. N. Thatoi (2017)
Copper (II) binding of NAD(P)H- flavin oxidoreductase (NfoR) enhances its Cr (VI)-reducing ability
Huawen Han (2017)
Development of bacterial nitroreductase enzymes for noninvasive imaging in cancer gene therapy
E. Williams (2013)
Bioremediation of hexavalent chromate using permeabilized Brevibacterium sp. and Stenotrophomonas sp. cells.
Shimei Ge (2015)
Plasmidic Expression of nemA and yafC* Increased Resistance of Ethanologenic Escherichia coli LY180 to Nonvolatile Side Products from Dilute Acid Treatment of Sugarcane Bagasse and Artificial Hydrolysate
A. Shi (2016)
Exploring novel Cr(VI) remediation genes for Cr(VI)-contaminated industrial wastewater treatment by comparative metatranscriptomics and metagenomics.
Yaxin Pei (2020)
Speciation, toxicity, microbial remediation and phytoremediation of soil chromium contamination
S. Guo (2020)
Hydrogen ions and organic acids secreted by ectomycorrhizal fungi, Pisolithus sp1, are involved in the efficient removal of hexavalent chromium from waste water.
L. Shi (2018)
Microbial interactions with chromium: basic biological processes and applications in environmental biotechnology
J. F. Gutiérrez-Corona (2016)
The use of bacterial bioremediation of metals in aquatic environments in the twenty-first century: a systematic review
Feliphe Lacerda Souza Alencar (2017)
Molecular mechanisms of Cr(VI) resistance in bacteria and fungi.
C. Viti (2014)
Bioengineering toward direct production of immobilized enzymes: A paradigm shift in biocatalyst design
Fabian B. H. Rehm (2018)
Mechanisms of Cr(VI) reduction by Bacillus sp. CRB-1, a novel Cr(VI)-reducing bacterium isolated from tannery activated sludge.
Y. Zhu (2019)
Alginate Encapsulation of Bioengineered Protein‐Coated Polyhydroxybutyrate Particles: A New Platform for Multifunctional Composite Materials
Kampachiro Ogura (2019)
Reduction of chromium-VI by chromium-resistant Escherichia coli FACU: a prospective bacterium for bioremediation
M. Mohamed (2020)
Hazards, Risks, and Low Hazard Development Paths of Synthetic Biology
B. Giese (2015)
Creation and screening of a multi-family bacterial oxidoreductase library to discover novel nitroreductases that efficiently activate the bioreductive prodrugs CB1954 and PR-104A.
G. Prosser (2013)
Polyhydroyxalkanoate Synthase Fusions as a Strategy for Oriented Enzyme Immobilisation
D. Hooks (2014)
Enzyme Engineering for In Situ Immobilization
Fabian B. H. Rehm (2016)
Recombinant surface engineering to enhance and expand the potential of biologically produced nanoparticles: A review
Gurusamy Kutralam-Muniasamy (2017)
Nitroreductase gene-directed enzyme prodrug therapy: insights and advances toward clinical utility.
E. Williams (2015)
Reduction of Hexavalent Chromium Using Recyclable Pt/Pd Nanoparticles Immobilized on Procyanidin-Grafted Eggshell Membrane
M. Liang (2014)
Multi-omics response of Pannonibacter phragmitetus BB to hexavalent chromium.
Li-yuan Chai (2019)
Bioremediation of Hexavalent Chromium Pollution by Sporosarcina saromensis M52 Isolated from Offshore Sediments in Xiamen, China.
R. Zhao (2016)
Simultaneous aerobic denitrification and Cr(VI) reduction by Pseudomonas brassicacearum LZ-4 in wastewater.
X. Yu (2016)
See more
Semantic Scholar Logo Some data provided by SemanticScholar