Genetic influence on biosynthesis of astaxanthin in Phaffia rhodozyma  Page description

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Details of project

  
Identifier
37471
Type K
Principal investigator Vágvölgyi, Csaba
Title in Hungarian Astaxanthin bioszintézis genetikai befolyásolása Phaffia rhodozyma-ban
Title in English Genetic influence on biosynthesis of astaxanthin in Phaffia rhodozyma
Panel Genetics, Genomics, Bioinformatics and Systems Biology
Department or equivalent Department of Microbiology (University of Szeged)
Participants Kasza, Zsolt
Nyilasi, Ildikó
Starting date 2002-01-01
Closing date 2005-12-31
Funding (in million HUF) 11.280
FTE (full time equivalent) 0.00
state closed project





 

Final report

  
Results in Hungarian
Az astaxanthin (3,3¢-dihydroxy-b,b-carotene-4,4¢-dione) termelő élesztők nagy biotechnológiai jelentőségűek. RAPD analízisen alapuló csoport analízist végrehajtva igazoltuk , hogy az astaxanthin-termelő izolátumok két fajba (Xanthopyllomyces dendrorhous és Phaffia rhodozyma) tartoznak. Különböző mutációs eljárásokat - gamma-, UV-sugárzás és kémiai mutagenezis, illetve ezek kombinációi - alkalmazva auxotróf és színmutánsokat izoláltunk mindkét élesztőfajból. Pigment- illetve astaxanthin túltermelő mutánsokat, melyek színanyag profilja jelentősen eltért hoztunk létre. Protoplaszt fúziós kísérleteket végeztünk és fúziós hibrideket izoláltunk és jellemeztünk. In vivo EPR vizsgálatokkal jellemeztük a különböző pigment-profilú mutánsok plazmamembránjának szerkezeti és dinamikus sajátosságait a hőmérséklettel összefüggésben. Meghatároztuk a X. dendrorhous 3-hydroxy-3-methylglutharyl coenzyme A reductase (HMG-CoA reductase) génjének részleges génszekvenciáját. Felfedeztük, hogy mind a P. rhodozyma és a X. dendrorhous képes lebontani az ochratoxin A-t ochratoxin alfává, feltehetően egy sejthez kötött karboxipeptidáz A aktivitás révén.
Results in English
Astaxanthin (3,3¢-dihydroxy-b,b-carotene-4,4¢-dione) - accumulating yeasts are of great biotechnological interest. RAPD analysis involving astaxanthin-producing yeast strains was performed: cluster analysis based on RAPD markers differentiated isolates of Xanthopyllomyces dendrorhous and Phaffia rhodozyma (asexual) at an intraspecific level. Different mutation strategies, which involves *- and UV-irradiations, as well as chemical mutagenesis of cells was worked out to obtain auxotrophic and colour mutants from these astaxanthin-producing yeasts. Overproducer mutants exhibited different patterns of carotenoid production have also been isolated. Protoplast fusion experiment have been carried out and fusion hybrides have been isolated and characterized. Spin labelling methods were used to describe the structure and dynamic properties of various carotenoids containing X. dendrorhous’s plasma membranes in relation to temperature. A partial gene sequence of 3-hydroxy-3-methylglutharyl coenzyme A reductase (HMG-CoA reductase) of X. dendrorhous has been determined. Ochratoxin degrading activities of P. rhodozyma and X. dendrorhous isolates were discovered. P. rhodozyma is able to convert ochratoxin A to ochratoxin a (possibly mediated by a cell-bound carboxypeptidase A).
Full text http://real.mtak.hu/220/
Decision
Yes





 

List of publications

  
Varga, J., Péteri, Zs., Tábori, K., and Vágvölgyi, Cs.: Degradation of ochratoxin A and other mycotoxins by Rhizopus isolates., Int. J. Food Microbiol. 99, 321-328., 2005
Ács, K., Kasza, Zs., Lukács, Gy., Schwab, H. and Vágvölgyi, Cs.: Cloning and sequence analysis of Mucor circinelloides glyceraldehyde-3-phosphate dehydrogenase gene, Acta Microbiol. Immunol. Hung. 49, 305-312., 2002
Nyilasi, I., Ács, K., Lukács, Gy., Papp, T., Kasza, Zs., and Vágvölgyi, Cs.: Agrobacterium tumefaciens-mediated transformation of Mucor circinelloides, FEMS Microbiol. Letters 222, Suppl. 1. 458., 2003
Lukács, Gy., Ács, K., Vastag, M., and Vágvölgyi, Cs.: Molecular cloning and sequence analysis of the gene encoding 3-hydroxy-3-methylglutharyl coenzyme A reductase in Rhizomucor miehei, FEMS Microbiol. Letters 222, Suppl. 1. 457., 2003
Nagy, Á., Pesti, M., Galgóczy, L. and Vágvölgyi, Cs.: Electrophoretic karyotype of two Micromucor species, J. Basic Microbiol. 44, 36-41., 2004
Vágvölgyi, Cs., Heinrich, H., Ács, K. and Papp, T.: Genetic variability in the species Rhizopus stolonifer, assessed by random amplified polymorphic DNA analysis, Antonie van Leeuwenhoek 86, 181-188., 2004
Vastag, M., Kasza, Zs, Ács, K., Papp, T., Schwab, H., and Vágvölgyi, Cs.: Cloning and sequences analysis of the glyceraldehyde-3-phosphate dehydrogenase gene from the zygomycetes fungus Rhizomucor miehei, Antonie van Leeuwenhoek 86, 111-119., 2004
Lukács, Gy., Papp, T., Nyilasi, I., Nagy, E., and Vágvölgyi, Cs.: Evaluation a new medium to differentiate Rhizomucor species on the basis of their different sensitivity to lovastatin., J. Clin. Microbiol. 42, 5400-5402., 2004
Palágyi, Zs., Papp, T., Takó, M., Nagy, Á., Pesti, M., and Vágvölgyi, Cs.: Genetic variability of astaxanthin-producing yeasts: random amplified polymorphic DNA (RAPD) analysis of Phaffia rhodozyma and Xanthopyllomyces dendrorhous, Acta Biol. Szeged. 48, 35-38., 2004
Papp, T., Eslava, A., Bartók, T., Vágvölgyi, Cs., and Iturriaga, E.: Heterologous expression of astaxanthin biosynthesis genes in Mucor circinelloides, Appl. Microbiol. Biotechnol. 69, 526-531., 2005
Lukács, Gy., Kovács, N., Papp, T., and Vágvölgyi, Cs.: The effect of vegetable oils on carotenoid production of Phaffia rhodozyma, Acta Microbiol. Immunol. Hung. 52, 267., 2005
Bartók, T., Papp, T., Lukács, Gy., Vágvölgyi, Cs.: High pressure liquid chromatography – atmospheric pressure photoionization mass spectrometry as a useful tool for the determination of carotenoids, Journal of Chromatography (submitted)., 2005
Takó, M., Vágvölgyi, Cs., Palágyi, Zs., Linka, B., and Papp, T.: Characterization of carotenoid over-producing Xanthophyllomycesdendrorhous mutants., Acta Microbiol. Hung. 52 (Suppl.) 161., 2005
Palágyi, Zs., Linka, B., Papp, T., and Vágvölgyi, Cs.: Isolation and characterization of Xanthophyllomyces dendrorhous mutants with altered carotenoid content., Acta Alimentaria Hung. (in press), 2006
Nyilasi, I., Ács, K., Papp, T., and Vágvölgyi, Cs.: Agrobacterium tumefaciens-mediated transformation of Mucor circinelloides, Folia Microbiol. 50, 414-420., 2005
Péteri, Zs., Téren, J., Vágvölgyi, Cs., and Varga, J.: Ochratoxin degradation and adsorption caused by astaxanthin-producing yeasts, Food Microbiol. (submitted), 2006
Csernetics, Á., Papp, T., Velayos, A., Iturriaga, E.A., Eslava, A.P., and Vágvölgyi, Cs.: Carotene production with genetically modified Mucor circinelloides strains., Acta Microbiol. Hung. 52 (Suppl.) 22., 2005
Papp, T., Csernetics, Á., Velayos, A., Iturriaga, E.A., Eslava, A.P., and Vágvölgyi, Cs.: Over-expression of isoprene biosynthetic enzymes in the b-carotene producer zygomycetes Mucor circinelloides., J. Biotech. 118 (S1), 153., 2005
Blasko, A., Belagyi, J., Dergez, T., Deli, J., Papp, G., Vagvolgyi, Cs. and Pesti, M.: Effect of the altered carotenoids composition on the plasma membrane dynamics of Xanthophyllomyces dendrorhous: in vivo EPR study., BBRC (submitted), 2006
Vágvölgyi, Cs., Lukács, Gy., Takó, M., Csernetics, Á., and Papp, T.: The effect of vegetable oils on astaxanthin production of Phaffia rhodozyma and Xanthophyllomyces dendrorhous, J. Biotech. 118 (S1), 153., 2005



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