A5025746420- Porphyrin Metabolism and Disorders
- Photosynthetic Processes and Mechanisms
- Enzyme Structure and Function
- Folate and B Vitamins Research
- Biochemical and Molecular Research
- Heme Oxygenase-1 and Carbon Monoxide
- Analytical Chemistry and Chromatography
- Metal-Catalyzed Oxygenation Mechanisms
- Enzyme Production and Characterization
- Neonatal Health and Biochemistry
- ATP Synthase and ATPases Research
- Algal biology and biofuel production
- Force Microscopy Techniques and Applications
- Vanadium and Halogenation Chemistry
- Advanced Electron Microscopy Techniques and Applications
- Microbial Metabolic Engineering and Bioproduction
- Iron Metabolism and Disorders
- Hemoglobin structure and function
- Porphyrin and Phthalocyanine Chemistry
- Metabolism and Genetic Disorders
- Electrochemical sensors and biosensors
- Eicosanoids and Hypertension Pharmacology
- Electron Spin Resonance Studies
- Mitochondrial Function and Pathology
- Photoreceptor and optogenetics research
University of Sheffield
2014-2025
University of Kent
2006-2010
Universidade Nova de Lisboa
2008
Lancaster City Council
2007
Queen Mary University of London
2000-2005
University of Exeter
1998-2004
University of York
2004
National Institute of Standards and Technology
2004
National Institutes of Health
2004
Many heterotrophic bacteria have the ability to make polyhedral structures containing metabolic enzymes that are bounded by a unilamellar protein shell (metabolosomes or enterosomes). These bacterial organelles contain associated with specific process (e.g. 1,2-propanediol ethanolamine utilization). We show 21 gene regulon specifying pdu organelle and propanediol utilization from Citrobacter freundii is fully functional when cloned in Escherichia coli, both producing metabolosomes allowing...
Reaction center-light harvesting 1 (RC-LH1) complexes are the fundamental units of bacterial photosynthesis, which use solar energy to power reduction quinone quinol prior formation proton gradient that drives ATP synthesis. The dimeric RC-LH1-PufX complex Rhodobacter sphaeroides is composed 64 polypeptides and 128 cofactors, including 56 LH1 bacteriochlorophyll a (BChl a) molecules surround donate two RCs. 3D structure was determined 8 Å by X-ray crystallography, model built with...
Abstract Inorganic phosphate is the major bioavailable form of essential nutrient phosphorus. However, concentration in most natural habitats low enough to limit microbial growth. Under phosphate-depleted conditions some bacteria utilise phosphite and hypophosphite as alternative sources phosphorus, but molecular basis reduced phosphorus acquisition from environment not fully understood. Here, we present crystal structures ligand binding affinities periplasmic proteins bacterial ATP-binding...
Escherichia coli has been engineered to produce chlorophyll.
The cobaltochelatase required for the synthesis of vitamin B12 (cobalamin) in archaeal kingdom has been identified as CbiX through similarity searching with from Bacillus megaterium. However, proteins archaea are much shorter than found eubacteria, typically containing less half number amino acids their primary structure. For this reason have termed CbiXS and longer versions CbiXL. Methanosarcina barkeri Methanobacter thermoautotrophicum were overproduced Escherichia coli recombinant...
The class II chelatases associated with heme, siroheme, and cobalamin biosynthesis are structurally related enzymes that insert a specific metal ion (Fe 2+ or Co ) into the center of modified tetrapyrrole (protoporphyrin sirohydrochlorin). structures two enzymes, CbiX S from Archaeoglobus fulgidus CbiK Salmonella enterica , responsible for insertion cobalt along pathway presented in complex their metallated product. A further structure Desulfovibrio vulgaris Hildenborough reveals how is...
Quantifying cellular components is a basic and important step for understanding how cell works, it responds to environmental changes, re-engineering cells produce valuable metabolites increased biomass. We quantified proteins in the model cyanobacterium Synechocystis sp. PCC 6803 given general importance of cyanobacteria global photosynthesis, synthetic biology biotechnology research, their ancestral relationship chloroplasts plants. Four mass spectrometry methods were used quantify involved...
The ability of Porphyromonas gingivalis to biosynthesize tetrapyrroles de novo has been investigated. Extracts the bacterium do not possess activity for 5- aminolevulinic-acid dehydratase or porphobilinogen deaminase, two key enzymes involved in synthesis uroporphyrinogen III. Similarly, it was possible detect any genetic evidence these early with use degenerate polymerase chain reaction. However, does appear harbor some cobalamin biosynthesis since cobyric acid, a pathway intermediate,...
The cDNAs of lactate dehydrogenase b (LDH-b) from both deep-sea and shallow living fish species, Corphaenoides armatus Gadus morhua respectively, have been isolated, sequenced their encoded products overproduced as recombinant enzymes in E. coli. proteins were characterised terms kinetic physical properties ability to withstand high pressures. Although the two are very similar primary structure, only 21 differences at amino acid level exist between them, enzyme species has a significantly...
The biosynthesis of the tetrapyrrole framework has been investigated in sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough by characterization enzymes required for transformation aminolaevulinic acid into sirohydrochlorin. PBG (porphobilinogen) synthase (HemB) was found to be a zinc-dependent enzyme that exists its native state as homohexamer. deaminase (HemC) shown contain dipyrromethane cofactor. Uroporphyrinogen III is fused with uroporphyrinogen methyltransferase...
The mature architecture of the photosynthetic membrane purple phototroph Rhodobacter sphaeroides has been characterised to a level where an atomic-level model is available, but roles putative assembly proteins LhaA and PucC in establishing this are unknown. Here we investigate light-harvesting LH2 reaction centre-light-harvesting1-PufX (RC-LH1-PufX) photosystem complexes using spectroscopy, pull-downs, native gel electrophoresis, quantitative mass spectrometry fluorescence lifetime...
Magnesium chelatase initiates chlorophyll biosynthesis, catalysing the MgATP2--dependent insertion of a Mg2+ ion into protoporphyrin IX. The catalytic core this large enzyme complex consists three subunits: Bch/ChlI, Bch/ChlD and Bch/ChlH (in bacteriochlorophyll producing species, respectively). D I subunits are members AAA+ (ATPases associated with various cellular activities) superfamily enzymes, they form that binds to H, site metal insertion. In order investigate physical coupling...
Abstract The biogenesis of Photosystem II is a complicated process requiring numerous auxiliary factors to assist in all steps its assembly. cyanobacterial protein Ycf39 forms stress-induced complex with two small chlorophyll-binding, High-light-inducible proteins C and D (HliC HliD), has been reported participate the insertion chlorophyll molecules into central D1 subunit II. However, how this organized remains unknown. Here, we show that both HliC HliD can form distinct complexes synthase...
NirJ is involved in the transformation of precorrin‐2 into heme d 1 , although its precise role process has not been established. The purified protein was found to contain a 4Fe–4S centre, line with prediction that it belongs radical SAM class enzymes. This further confirmed by binding S‐adenosyl‐ l ‐methionine (SAM) dithionite‐reduced NirJ, which resulted decrease signal intensity and shift higher field [4Fe–4S] 1+ EPR signal. Significantly, though, this approach also led appearance small...
The sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough possesses a large number of porphyrin-containing proteins whose biosynthesis is poorly characterized. In this work, we have studied two putative CbiK cobaltochelatases present in the genome D. vulgaris. assays revealed that both enzymes insert cobalt and iron into sirohydrochlorin, with specific activities lower than measured cobalt. Nevertheless, chelatases complement an E. coli cysG mutant strain showing that, vivo, they...
Ring contraction during cobalamin (vitamin B12) biosynthesis requires a seemingly futile methylation of the C20 position tetrapyrrole framework. Along anaerobic route, this reaction is catalyzed by CbiL, which transfers methyl group from S-adenosyl-L-methionine to cobalt factor II generate III. CbiL belongs class III methyltransferases and displays similarity other biosynthetic that are responsible for regiospecific number positions on molecular canvas. In an attempt understand how...