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Contact Details Department of Biology (Area 8) University of York PO Box 373 York YO10 5YW Tel: +44 (0)1904 328777 E-mail: ncb5@york.ac.uk Office: B/M219 Web Pages: CNAP Bruce Lab web pages

Career Outline

1983	BSc Applied Biology	Hatfield Polytechnic
1986-Sep 90	Postdoctoral Research Associate	Institute of Biotechnology, University of Cambridge
1987	PhD	University of Kent
1988-Sep 91	Research Fellow	Wolfson College, Cambridge
1990-Sep 01	University Lecturer	Institute of Biotechnology, University of Cambridge
1991-Sep 02	Staff Fellow, Tutor and Director of Studies for Natural Sciences (Biological)	Trinity Hall, Cambridge
2001-Jun 02	Reader in Biotechnology	University of Cambridge
2002-	Professor of Biotechnology	University of York

Research Interests

The major research themes of our laboratory are microbial metabolism, biocatalysis and environmental biotechnology. A primary goal is to understand how microorganisms have adapted to utilise xenobiotic compounds as carbon and nitrogen sources for growth. The enzymes mediating these pathways often have potential commercial applications as recognition components in biosensors, as biocatalysts for synthetic chemistry and for the bioremediation of soil and ground water. We are now engaged in extensive structural analysis of a number of these enzymes using X-ray crystallography. Work is also focusing on generating carefully designed mutant forms of a number of these enzymes to understand their catalytic mechanisms. A principal theme of our research is the biodegradation, biotransformation and phytoremediation of explosives.

Discoveries
We discovered a novel cytochrome P450 system termed XplA/B from Rhodococcus rhodochrous (11Y) that degrades the high explosive RDX. As a model system for RDX phytoremediation, Arabidopsis expressing XplA/B were grown in RDX contaminated soil and found to remove and degrade the explosive from the soil. Our work suggests that expressing XPlA/B in landscape plants may provide a suitable remediation strategy for explosive contaminated sites.

Some Recent Publications

Jackson RG, Rylott EL, Fournier D, Hawari J and Bruce NC (2007) Exploring the biochemical properties and remediation applications of the unusual explosive degrading cytochrome P450 system XplA/B Proc. Natl. Acad. Sci. USA 104: 16822-827

Gandia-Herrero F, Lorenz A, Larson TR, Graham IA, Bowles DJ, Rylott EL, Bruce NC (2008) Detoxification of the explosive 2,4,6-trinitrotoluene in Arabidopsis- discovery of bifunctional O- and C-glucosyltransferases. The Plant Journal 56: 963-974.

Szolkowy, C., Eltis, L.D., Bruce, N.C. and Grogan, G. (2009) Insights into sequence-activity relationships amonst Baeyer-Villiger Monooxygenases as revealed by the intragenomic complement of enzymes from Rhodococcus jostii RHA1 ChemBioChem 10: 1208-1217.

Current Research Projects

• Advanced optical waveguide biosensors for the detection of illicit drugs and explosives (joint with Nicholas Goddard, University of Manchester)
  Funding body: BBSRC
• Sustainable range management of RDX and TNT by phytoremediation with engineered plants
  Funding body: SERDP
• Engineering cytochrome P450s for biohydroxylation (joint with Gideon Grogan, YSBL)
  Funding body: CoEBio3
• Gold phytomining
  Funding body: Burgess
• Rhizosphere bacterial degradation of RDX, Understanding and enhancement
  Funding body: SERDP
• The molecular biology of nitroamine degradation in soils
  Funding body: SERDP
• Developing new enzymes for novel bioprocesses
  Funding body: CoEBio3, Manchester
• Engineering plants for the phytoremediation of TNT
  Funding body: BBSRC
• Developing a new platform technology for oxidative biocatalysis. Mining the potential of plant P450s for biohydroxylations
  Funding body: CoEBio3
• New tools for the realisation of cost-effective liquid biofuels from plant biomass (joint with Simon McQueen-Mason)
  Funding body: BBSRC
• Engineering cytochrome P450s for biohydroxylation (joint with Gideon Grogan, Department of Chemistry)
  Funding body: CoEBio3

Professional Activities

• Editorial Boards: Applied and Environmental Microbiology
• Member of the College of Reviewers for the Canada Research Chairs Program
• York Science Director of CoEBio3 (Centre of Excellence in Biocatalysis, Biotransformations and Biocatalytic Manufacture)
• Co-Founder and Director of Bioniqs Ltd

PhD Research Projects Available for 2010

Exploring the molecular biology of gold uptake in plants (for 2010-11)
Large amounts of land worldwide are unfit for agricultural use as a result metal mining. Interestingly, some plants have the remarkable ability to hyperaccumulate these toxic metals, a property which has only recently been exploited commercially to phytomine nickel. To realise the full potential of this technology, a significant amount of work is now focused on understanding the molecular mechanisms behind these processes. In field trials conducted at former mines in Brazil, plants were shown to accumulate mercury, the toxic metal used by miners in the past to extract gold. Surprisingly, these plants also accumulated gold. More recent studies have found that plants such as oilseed rape, corn and mustard planted on old mining tailings with high concentrations of gold take up solubilised gold storing it in the cell walls as gold particles. Although the process requires the use of low concentrations of cyanide to solubilise the gold, the process is controlled and results in an improvement of the quality of the soil at the end of the treatment. The gold is easily recoverable following incineration; providing a cash incentive for miners to clean their land, re-green mining sites and cleaning waterways. We are currently exploring the biology underpinning gold uptake and deposition in plants. We have observed gold uptake in Arabidopsis, a relative of oilseed rape and mustard, using liquid culture studies and electron microscopy. The aim of this project is to determine the genes, proteins and mechanisms involved these processes in the plant Arabidopsis. The project will make use of transgenic tools, mutant screens and genomics data to elucidate the molecular biology and biochemistry involved in the initial response and subsequent translocation, processing and storage of gold in plants.

Engineering explosives degrading enzymes (for 2010-11)
Royal Demolition Explosive (RDX) and 2,4,6-trinitrotoluene (TNT) are widely used military explosives and are environmental hazards owing to their toxicity and recalcitrance to biodegradation. Concern is growing regarding the large areas of land and ground water polluted worldwide as a result of the manufacture, use and disposal of these compounds. We have a long-standing interest in discovering how microorganisms and plants metabolise xenobiotic compounds such as explosives. This project will focus on two enzyme systems: an unusual bacterial cytochrome P450, XplA, that can degrade RDX and plant glutathione transferases that possess activity towards TNT. We have heterologously expressed and purified both XplA and the TNT-active GSTs. Interestingly, recombinant XplA was shown to degrade RDX both aerobically and anaerobically by a reductive denitration mechanism. The presence of oxygen appears to determine the final products of RDX degradation, demonstrating that the degradation chemistry is flexible. A reductive denitration mechanism is unusual for P450s and the details of this are still unclear. We have recently solved the structure of the heme domain of XplA has been solved in two crystal forms. Like RDX, TNT is highly resistant to biodegradation; however we have shown that certain plant GSTs apparently catalyse a novel denitration of TNT resulting in a transformation product that may be more amenable to microbial degradation. The way is now open for further characterisation, optimisation and engineering of these environmentally important enzymes.

Lab Members

Status	Name	Project
Admin	Margaret Cafferky
Post Doctoral Fellow	Dr Hazel Housden	Biosensor development for the detection of explosives and illicit drugs
Post Doctoral Fellow	Joe Bennett	Developing new enzymes for novel bioprocesses
Post Doctoral Fellow	Dr Katrin Besser	Discovering novel lignocellulose degrading enzymes from 'gribble' for the production of liquid biofuels (joint supervision with Simon McQueen-Mason)
Post Doctoral Fellow	Dr Marcelo Kern	Discovering novel lignocellulose degrading enzymes from 'gribble' for the production of liquid biofuels (joint supervision with Simon McQueen-Mason)
Post Doctoral Fellow	Dr Astrid Lorenz	Rhizosphere bacterial degradation of RDX, Understanding and enhancement
Post Doctoral Fellow	Dr Elizabeth Rylott	Sustainable range management of RDX and TNT by phytoremediation
Post Doctoral Fellow	Dr Cyril Bontemps	The molecular biology of nitroamine degradation in soils
Research Student	Claudia Szolkowy	Developing a generic high-throughput method for the rapid generation of 'designer' oxidoreductase biocatalysts (joint supervision with Gideon Grogan, YSBL)
Research Student	Julia Schückel	Developing a new platform technology for oxidative biocatalysis: Mining the potential of plant P450s for biohydroxylations
Research Student	William Eborall	Discovering novel lignocellulose degrading enzymes from ‘gribble’ for the production of liquid biofuels (joint supervision with Simon McQueen-Mason)
Research Student	Federico Sabbadin	Engineering cytochrome P450s for biohydroxylation
Research Student	Helen Sparrow	Engineering plants for the phytoremediation of TNT
Research Student	Andrew Taylor	Gold phytomining
Research Student	Chung Shion Chong	Microbial degradation of the explosive RDX
Research Student	Florian Fisch	New malonate decarboxylases for industrial biocatalysis (joint supervision with Gideon Grogan, YSBL)
Technician	Luisa Elias	Discovering novel lignocellulose degrading enzymes from 'gribble' for the production of liquid biofuels (joint supervision with Simon McQueen-Mason)
Visitor	Eric Nassau	microbial degradation of Endocrine Disrupting Compounds (Graduate student, University of Washington, USA).