Description of project
In the proposed master project, you will be working on the enzyme hydrogenase, which functionally catalyze the hydrogen gas production from proton H+ and electron donor. You will design mutants of hydrogenases in order to increase the thermostability, the catalytic activity and reduce the oxygen sensitivity of the enzyme. You will transform plasmid DNA constructs with mutant hydrogenases and induce protein expression in the host bacteria E.coli. The overexpressed recombinant protein will be purified in large scale. The study will use protein structure analysis tools in the design process. In addition, with the similar strategy, you will take part in cloning, expression and purification of the enzyme formate dehydrogenase, aiming to be used as biocatalyst to capture environment CO2 and produce formic acid.
Methods
During this project you will gain work experience in a wide variety of practical techniques, including recombinant DNA technology, protein structure, PCR, protein expression and purification, protein biochemistry, enzymatic assays, Western blotting and related nanotechnology.
Research environment
We have purified and biochemically characterized a number of recombination proteins, including the DNA/RNA repair enzymes endonuclease V (Dalhus et al, Nature Struct Mol Biol. 2009; Vik ES et al, Nature Commun. 2013), the DNA glycosylase OGG1 (Dalhus B et al, Structure. 2011) and a new HEAT-like repeat DNA glycosylase involved in recombination repair (Backe PH et al J Struct Biol. 2013). In cell biology, we have identified and characterized a novel anti-oxidation protein OXR1. By knocking down in cell lines, we showed that it functionally prevents oxidative stress induced cell death and maintains genome stability by up-regulating an anti-oxidation pathway (Yang et al, Free Radic Biol Med. 2014; Scientific reports 2015).
Working place
Oslo University Hospital - Rikshospitalet, Department for Medical Biochemistry.
Contact persons
Dr Paul Hoff Backe, Department for Medical Biochemistry, Oslo University Hospital.
Dr Bj?rn Dalhus, Department for Medical Biochemistry, Oslo University Hospital
Dr Mingyi Yang, Department for Microbiology, Oslo University Hospital