School of Biological Sciences
Royal Holloway University of London
Egham, Surrey, TW20 0EX
United Kingdom

Tel: 01784-443555
Fax: 01784-414224
E-mail:p.bramley@rhul.ac.uk

Carotenoids are a group of major plant pigments responsible for most of the yellow to red colours of flowers, fruits and vegetables. In the diet they act as powerful antioxidants and are believed to protect the body against free radical attack and hence reduce the incidence of cataracts, heart disease and certain cancers.

One approach to increasing the dietary intake of carotenoids is to increase their levels in fruit and vegetables. In order to achieve this goal, it Is necessary to understand how plants control the formation and accumulation of carotenoids, using biochemical, molecular biology and electron microscopy techniques. Currently, we are focusing on the tomato in order to elevate the levels of the three major carotenoids in ripe fruit, lycopene, ß-carotene and astaxanthin.

Carotenoid genes isolated from plants and bacteria and encoding key enzymes in the pathway have been introduced into the tomato by genetic transformation using Agrobacterium-based vectors. The transgenic plants are analysed for pigment content by HPLC, gene expression (northern and western blotting, RT-PCR), plastid ultrastructure and antioxidant potential. We are also analysing a large number of introgression lines of tomato by metabolomics in order to identify quantitative trait loci (QTLs) that can be used in plant breeding to produce elite lines that have elevated levels of antioxidants.

We are also developing a systems biology approach to predicting the metabolic control of isoprenoid biosynthesis and accumulation in tomato.

We are using proteomics (MudPIT, HPLC, IEX, SEC, electrophoresis, isoelectric focusing) and metabolomics (GC/MS, HPLC/MS, HPLC/DAD) in studies to detect:

• False labelling of foods
• Adulteration of foods
• Toxic compounds in foods
• GM ingredients in foods and feeds.
• Identification of animal proteins in feeds.

• Carotenoid biosynthesis and its regulation in transgenic and mutant varieties of tomato, funded by Syngenta, EU (EU-Sol) and BBSRC.
• A systems biology approach to the elucidation of metabolic networks underlying health based quality traits in tomato fruit, funded by BBSRC as part of ANR/BBSRC projects in systems biology.
• Development of bioinformatic software (BioSynLab) with colleagues in the Department of Computer Science
• Development of high throughput approaches to optimise the nutritional value of crops and crop-based foods. Funded by EU, in Develonutri network.
• New sources of natural, gastric stable, food additives colourants and novel functional foods. Funded by EU, in Colorspore network.
• Metabolomic approaches to the identification of health based consumer traits in tomato, funded by BBSRC
• The rapid and quantitative estimation of gelatine-derived peptides in animal feed, funded by Food Standards Agency
• The application of metabolic profiling to the safety assessment of GM foods, funded by the Foods Standards Agency.
• The development of sensitive and robust mass spectrometric methods for the detection of GM material in foods and food products, funded by the Foods Standards Agency
• A bioinformatic approach to the elucidation of regulators involved in the formation of health promoting phytochemicals, funded by the British-Italian Councils.
• South Africa-UK exploratory network, funded by the Royal Society.
• A systems biology approach to tomato fruit ripening. (BBSRC-ESB-LINK) with Prof. Graham Seymour, University of Nottingham and Syngenta.
• The application of mass spectrometry to the identification and characterisation of animal proteins in foods (BBSRC CASE award with PDM Foods).
• Metabolite profiling of fungi for long chain fatty polyunsaturated fatty acids and carotenoids (with CABI)
• A systems biology approach to the study of flavonoid biosynthesis in tomato (CASE award with Syngenta).
• Metagenomic approaches to study intervention strategies for Salmonella typhimurium in pigs (with VLA Weybridge), funded by FSA
• Comparative genomics to identify genetic variabilities associated with Salmonella typhimurium DT104 epidemicity (with VLA Weybridge)
• HPLC/mass spectrometry methods for the detection of active
• Immunogens in Leptospira vaccine batches. With VLA Weybridge.
• Genetic variability within Salmonella enterica serovar enteritidis can be used to identify markers associated with human epidemicity (with VLA Weybridge)

• Automated HPLC-diode array spectrophotometry with on-line radioactivity detector for carotenoid analyses.
• Analysis of sterols and tocopherols by GC and HPLC.
• Metabolite profiling using HPLC, GC/MS and LC/MS/MS.
• Proteomics using 2D-PAGE, multidimensional chromatography and MS/MS
• Characterisation of carotenogenic enzymes in vitro
• Purification of carotenogenic enzymes.
• Molecular biology techniques on higher plants, e.g. gene cloning, mRNA analysis, quantitative RT-PCR, real time PCR, vector construction
• Transformation of tomato, tobacco and flax.

Mora L, Sentandreu M, Koistenen K, Fraser PD, Toldra F and Bramley PM. (2009) Peptide Fragments Naturally Generated in Dry-Cured Ham, J Agric Food Chem, in press

Fraser PD and Bramley PM. Genetic Manipulation of Carotenoid Content and Composition in Crop Plants. In Carotenoids, vol 5 (Britton G., ed), chapter 6, in press

Fernández Ocaña M, Paul D. Fraser PD, Patel RKP, Halket JM and Peter M. Bramley (2008) Stable isotope labelling strategies for the quantitation of CP4 EPSPS in genetically modified soya. Analytica Chim Acta. 2009 634(1): 75-82

Fraser PD, Enfissi EMA and Bramley PM. Genetic engineering of carotenoid formation in tomato fruit and the potential application of systems and synthetic biology approaches. Archiv Biochem Biophys, 2009 483(2): 196-204

Shindo K, Endo M, Miyake Y, Wakasugi K, Morritt D, Bramley PM, Fraser PD, Kasai H, Misawa N.(2008) Methyl glucosyl-3,4-dehydro-apo-8'-lycopenoate, a novel antioxidative glyco-C(30)-carotenoic acid produced by a marine bacterium Planococcus maritimus. J Antibiot (Tokyo) 2008 12:729-735

Dagustu N, Fraser PD, Enfissi EMA and Bramley PM (2008) Screening for high callus induction and Agrobacterium-mediated transformation of sunflower (Helianthus annuus l.) Biotechnology and Biotechnological Equipment, 2008 22(4): 933-937

Fraser PD, Enfissi EM, Halket JM, Truesdale MR, Yu D, Gerrish C, Bramley PM. Manipulation of phytoene levels in tomato fruit: effects on isoprenoids, plastids, and intermediary metabolism. Plant Cell. 2007 19(10):3194-211

Millar DJ, Long M, Donovan G, Fraser PD, Boudet AM, Danoun S, Bramley PM, Bolwell GP. Introduction of sense constructs of cinnamate 4-hydroxylase (CYP73A24) in transgenic tomato plants shows opposite effects on flux into stem lignin and fruit flavonoids. Phytochemistry. 2007 68(11):1497-509

Simkin AJ, Gaffé J, Alcaraz JP, Carde JP, Bramley PM, Fraser PD, Kuntz M. Fibrillin influence on plastid ultrastructure and pigment content in tomato fruit. Phytochemistry. 2007 68(11):1545-56

Fraser PD, Enfissi EM, Goodfellow M, Eguchi T, Bramley PM. Metabolite profiling of plant carotenoids using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Plant J. 2007 49(3):552-64

Ocaña MF, Fraser PD, Patel RK, Halket JM, Bramley PM. Mass spectrometric detection of CP4 EPSPS in genetically modified soya and maize. Rapid Commun Mass Spectrom. 2007 21(3):319-28

Enfissi EM, Fraser PD, Lois LM, Boronat A, Schuch W, Bramley PM. Metabolic engineering of the mevalonate and non-mevalonate isopentenyl diphosphate-forming pathways for the production of health-promoting isoprenoids in tomato. Plant Biotechnol J. 2005 3(1):17-27

Xu CJ, Fraser PD, Wang WJ, Bramley PM. Differences in the carotenoid content of ordinary citrus and lycopene-accumulating mutants. J Agric Food Chem. 2006 54(15):5474-81

Long M, Millar DJ, Kimura Y, Donovan G, Rees J, Fraser PD, Bramley PM, Bolwell GP. Metabolite profiling of carotenoid and phenolic pathways in mutant and transgenic lines of tomato: identification of a high antioxidant fruit line. Phytochemistry. 2006 67(16):1750-7

Davuluri GR, van Tuinen A, Fraser PD, Manfredonia A, Newman R, Burgess D, Brummell DA, King SR, Palys J, Uhlig J, Bramley PM, Pennings HM, Bowler C. (2005) Fruit-specific RNAi-mediated suppression of DET1 enhances carotenoid and flavonoid content in tomatoes. Nat Biotechnol. 23: 890-5 [PubMed]

Fernandez Ocana M, Jarvis J, Parker R, Bramley PM, Halket JM, Patel RK, Neubert H. (2005) C-terminal sequencing by mass spectrometry: application to gelatine-derived proline-rich peptides. Proteomics. 5: 1209-16 [PubMed]

Giliberto L, Perrotta G, Pallara P, Weller JL, Fraser PD, Bramley PM, Fiore A, Tavazza M, Giuliano G. (2005) Manipulation of the blue light photoreceptor cryptochrome 2 in tomato affects vegetative development, flowering time, and fruit antioxidant content. Plant Physiol. 137: 199-208 [PubMed]

Halket JM, Waterman D, Przyborowska AM, Patel RK, Fraser PD, Bramley PM. (2005) Chemical derivatization and mass spectral libraries in metabolic profiling by GC/MS and LC/MS/MS. J Exp Bot. 56: 219-43 [PubMed]

Ralley L, Enfissi EM, Misawa N, Schuch W, Bramley PM, Fraser PD. (2004) Metabolic engineering of ketocarotenoid formation in higher plants. Plant J. 39: 477-86 [PubMed]

Fraser PD and Bramley PM (2004) The biosynthesis and nutritional uses of carotenoids. Prog. Lipid Res. 43: 228-265.

Fernandez-Ocana M, Neubert H, Przyborowska A, Parker R, Bramley PM, Halket J and Patel R (2004) BSE control: detection of gelatine-derived peptides in animal feed by mass spectrometry. Analyst 129: 111-115.

Giuliano G, Rosati C and Bramley PM (2003) To dye or not to dye: biochemistry of annatto unveiled. Trends Biotechnol. 21: 513-516

Bramley PM (2003) Genetic engineering for enhancing levels of carotenoids in plants. In: Plant Genetic Engineering, vol.1 (RP Singh and PK Jaiwal, eds), chaper 10, pp.229-259.

Cookson PJ, Kiano JW, Shipton CA, Fraser PD, Romer S, Schuch W, Bramley PM and Pyke KA (2003) Increases in cell elongation, plastid compartment size and phytoene synthase activity underlie the phenotype of the high pigment-1 mutant of tomato. Planta. 217: 896-903.

Bramley PM (2003) The genetic enhancement of phytochemicals: the case of carotenoids. In: Phytochemical Functional Foods (Johnson I and Williamson G, eds), chapter 13, pp. 253-0279. Woodhead Publishing Ltd, Cambridge.

Burns J, Fraser PD, Bramley PM (2003) Identification and quantification of carotenoids, tocopherols and chlorophylls in commonly consumed fruits and vegetables. Phytochemistry 62: 939-947.

Shahmuradov IA, Gammerman AJ, Hancock JM, Bramley PM, Solovyev VV (2003) PlantProm: a database of plant promoter sequences. Nucleic Acids Res. 31: 114-117. [Full text 76 KB]

Fester T, Schmidt D, Lohse S, Walter MH, Giuliano G, Bramley PM, Fraser PD, Hause B, Strack D (2002) Stimulation of carotenoid metabolism in arbuscular mycorrhizal roots. Planta 216: 148-154. [Full text 384 KB]

Fraser PD, Romer S, Shipton CA, Mills PB, Kiano JW, Misawa N, Drake RG, Schuch W, Bramley PM (2002) Evaluation of transgenic tomato plants expressing an additional phytoene synthase in a fruit-specific manner. Proc. Natl. Acad. Sci. USA 99: 1092-1097. [Full text 152 KB]

Re R, Bramley PM, Rice-Evans C (2002) Effects of food processing on flavonoids and lycopene status in a Mediterranean tomato variety. Free Radic. Res. 36: 803-810.

Fraser PD, Bramley P, Seymour GB (2001) Effect of the Cnr mutation on carotenoid formation during tomato fruit ripening. Phytochemistry 58: 75-79. [Full text 126 KB]

Fraser PD, Romer S, Kiano JW, Shipton CA, Mills PB, Drake R, Schuch W, Bramley PM (2001) Elevation of carotenoids in tomato by genetic manipulation. J. Sci. Food Agric. 81: 822-827

Re R, Fraser PD, Long M, Bramley PM, Rice-Evans C (2001) Isomerization of lycopene in the gastric milieu. Biochem. Biophys. Res. Commun. 281: 576-581

Fraser PD, Pinto MES, Holloway DE, Bramley PM (2000) Application of high-performance liquid chromatography with photodiode array detection to the metabolic profiling of plant isoprenoids. Plant J. 24: 551-558.

Fraser PD, Schuch W, Bramley PM (2000) Phytoene synthase from tomato (Lycopersicon esculentum) chloroplasts - partial purification and biochemical properties. Planta 211: 361-369.]

Romer S, Fraser PD, Kiano JW, Shipton CA, Misawa N, Schuch W, Bramley PM (2000) Elevation of the provitamin A content of transgenic tomato plants. Nat. Biotechnol. 18: 666-669

Bramley PM (2000) Is lycopene beneficial to human health? Phytochemistry 54: 233-236. [Full text 94 KB]

Bramley PM, Elmadfa I, Kafatos A, Kelly FJ, Manios Y, Roxborough HE, Schuch W, Sheehy PJA, Wagner KH (2000) Vitamin E. J. Sci. Food Agric. 80: 913-938.

Holloway DE, Yang M, Paganga G, Rice-Evans CA, Bramley PM (2000) Isomerization of dietary lycopene during assimilation and transport in plasma. Free Radic. Res. 32: 93-102.

Fraser PD, Kiano JW, Truesdale MR, Schuch W, Bramley PM (1999) Phytoene synthase-2 enzyme activity in tomato does not contribute to carotenoid synthesis in ripening fruit. Plant Mol.Biol. 40: 687-698.

Harker M, Bramley PM (1999) Expression of prokaryotic 1-deoxy-D-xylulose-5-phosphatases in Escherichia coli increases carotenoid and ubiquinone biosynthesis. FEBS Lett. 448: 115-119.[Full text 355 KB]

Fraser PD, Misawa N, Sandmann G, Johnson J, Schuch W, Bramley PM (1998) Production and characterisation of monoclonal antibodies to phytoene synthase of Lycopersicon esculentum. Phytochemistry 49: 971-978.

Kuntz M, Chen HC, Simkin AJ, Romer S, Shipton CA, Drake R, Schuch W, Bramley PM (1998) Upregulation of two ripening-related genes from a nonclimacteric plant (pepper) in a transgenic climacteric plant (tomato). Plant J. 13: 351-361.

Bramley PM (1997) The regulation and genetic manipulation of carotenoid biosynthesis in tomato fruit. Pure Appl. Chem. 69: 2159-2162.

RiceEvans CA, Sampson J, Bramley PM, Holloway DE (1997) Why do we expect carotenoids to be antioxidants in vivo? Free Radic. Res. 26: 381-398.

Schuch W, Drake R, Romer S, Bramley PM (1996) Manipulating carotenoids in transgenic plants, in Engineering Plants for Commercial Products and Applications, pp 13-19.

Miller NJ, Sampson J, Candeias LP, Bramley PM, RiceEvans CA (1996) Antioxidant activities of carotenes and xanthophylls. FEBS Lett. 384: 240-242.[Full text 282 KB]

Fraser PD, RuizHidalgo MJ, LopezMatas MA, Alvarez MI, Eslava AP, Bramley PM (1996) Carotenoid biosynthesis in wild type and mutant strains of Mucor circinelloides. Biochim. Biophys. Acta-Gen. Subj. 1289: 203-208.