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Nitroreductase from Salmonella typhimurium: characterization and catalytic activity
Yanto Yanto1, Mélanie Hall, Andreas S Bommarius
1School of Chemical and Biomolecular Engineering, Parker H. Petit Institute of Bioengineering and Biosciences, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA 30332-0363, USA.
Organic & Biomolecular Chemistry
|May 8, 2010
Summary
Salmonella typhimurium nitroreductase (NRSal) efficiently reduces nitroalkenes and nitroaromatics, offering a biocatalytic route to nitroalkanes and aniline. While enone reduction is limited, NRSal shows broad substrate scope and high conversion efficiency.
Area of Science:
- Biochemistry
- Enzymology
- Organic Synthesis
Background:
- Nitroreductases are crucial enzymes for reducing nitro-containing compounds.
- Investigating novel nitroreductases can expand biocatalytic toolkits for chemical synthesis.
Purpose of the Study:
- To characterize the biocatalytic activity of nitroreductase from Salmonella typhimurium (NRSal).
- To explore NRSal's potential in reducing alpha,beta-unsaturated carbonyl compounds, nitroalkenes, and nitroaromatics.
Main Methods:
- Gene synthesis and subcloning into a pET28 overexpression system.
- Expression in E. coli BL21 and purification of the NRSal protein.
- Assaying biocatalytic activity with various nitro-containing substrates.
Main Results:
- NRSal exhibited broad substrate acceptance for nitroalkenes and nitroaromatics (>95% conversion).
- High stereoselectivity was observed in the reduction of enones, though overall efficiency was low.
- NRSal catalyzed the reduction of nitrobenzene to aniline via nitrosobenzene and phenylhydroxylamine intermediates.
Conclusions:
- NRSal is an efficient biocatalyst for preparing nitroalkanes from nitroalkenes.
- The enzyme facilitates the first reported single enzyme-catalyzed reduction of nitrobenzene to aniline.
- Further optimization is needed to overcome intermediate condensation and improve aniline yield.
