Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous

The cloning and nucleotide sequence of the genes (idi, crtE, crtYB, crtl and crtS) controlling the astaxanthin biosynthesis pathway of the wild-type ATCC 24230 strain of Xanthophyllomyces dendrorhous in their genomic and cDNA version were obtained. The idi, crtE, crtYB, crtl and crtS genes were clon...

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Autores principales:	NIKLITSCHEK,MAURICIO, ALCAÍNO,JENNIFER, BARAHONA,SALVADOR, SEPÚLVEDA,DIONISIA, LOZANO,CARLA, CARMONA,MARISELA, MARCOLETA,ANDRÉS, MARTÍNEZ,CLAUDIO, LODATO,PATRICIA, BAEZA,MARCELO, CIFUENTES,VÍCTOR
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2008
Materias:	astaxanthin biosynthesis Xanthophyllomyces dendrorhous Phaffia rhodozyma
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100011
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spelling	oai:scielo:S0716-976020080001000112008-08-21Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhousNIKLITSCHEK,MAURICIOALCAÍNO,JENNIFERBARAHONA,SALVADORSEPÚLVEDA,DIONISIALOZANO,CARLACARMONA,MARISELAMARCOLETA,ANDRÉSMARTÍNEZ,CLAUDIOLODATO,PATRICIABAEZA,MARCELOCIFUENTES,VÍCTOR astaxanthin biosynthesis Xanthophyllomyces dendrorhous Phaffia rhodozyma The cloning and nucleotide sequence of the genes (idi, crtE, crtYB, crtl and crtS) controlling the astaxanthin biosynthesis pathway of the wild-type ATCC 24230 strain of Xanthophyllomyces dendrorhous in their genomic and cDNA version were obtained. The idi, crtE, crtYB, crtl and crtS genes were cloned, as fragments of 10.9, 11.5, 15.8, 5.9 and 4 kb respectively. The nucleotide sequence data analysis indicates that the idi, crtE, crtYB, crtl and crtS genes have 4, 8,4, 11, and 17 introns and 5, 9, 5, 12 and 18 exons respectively. In addition, a highly efficient site-directed mutagenesis system was developed by transformation by integration, followed by mitotic recombination (the double recombinant method). Heterozygote idi (idi+ / idi-::hph), crtE (crtE+ / crtE -::hph), crtYB (crtYB + / crtYB -::hph), crtI (crtI+ / crtI-::hph) and crtS (crtS +/crtS -::hph) and homozygote mutants crtYB (crtYB -::hph/crtYB -::hph), crtI (crtI -::hph/crtI -::hph) and crtS (crtS -::hph / crtS -::hph) were constructed. All the heterozygote mutants have a pale phenotype and produce less carotenoids than the wild-type strain. The genetic analysis of the crtYB, crtl and crtS loci in the wild-type, heterozygote, and homozygote give evidence of the diploid constitution of ATCC 24230 strains. In addition, the cloning of a truncated form of the crtYB that lacks 153 amino acids of the N-terminal region derived from alternatively spliced mRNA was obtained. Their heterologous expression in Escherichia coli carrying the carotenogenic cluster of Erwinia uredovora result in trans-complementation and give evidence of its functionality in this bacterium, maintaining its phytoene synthase activity but not the lycopene cyclase activityinfo:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.41 n.1 20082008-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100011en10.4067/S0716-97602008000100011
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	astaxanthin biosynthesis Xanthophyllomyces dendrorhous Phaffia rhodozyma
spellingShingle	astaxanthin biosynthesis Xanthophyllomyces dendrorhous Phaffia rhodozyma NIKLITSCHEK,MAURICIO ALCAÍNO,JENNIFER BARAHONA,SALVADOR SEPÚLVEDA,DIONISIA LOZANO,CARLA CARMONA,MARISELA MARCOLETA,ANDRÉS MARTÍNEZ,CLAUDIO LODATO,PATRICIA BAEZA,MARCELO CIFUENTES,VÍCTOR Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous
description	The cloning and nucleotide sequence of the genes (idi, crtE, crtYB, crtl and crtS) controlling the astaxanthin biosynthesis pathway of the wild-type ATCC 24230 strain of Xanthophyllomyces dendrorhous in their genomic and cDNA version were obtained. The idi, crtE, crtYB, crtl and crtS genes were cloned, as fragments of 10.9, 11.5, 15.8, 5.9 and 4 kb respectively. The nucleotide sequence data analysis indicates that the idi, crtE, crtYB, crtl and crtS genes have 4, 8,4, 11, and 17 introns and 5, 9, 5, 12 and 18 exons respectively. In addition, a highly efficient site-directed mutagenesis system was developed by transformation by integration, followed by mitotic recombination (the double recombinant method). Heterozygote idi (idi+ / idi-::hph), crtE (crtE+ / crtE -::hph), crtYB (crtYB + / crtYB -::hph), crtI (crtI+ / crtI-::hph) and crtS (crtS +/crtS -::hph) and homozygote mutants crtYB (crtYB -::hph/crtYB -::hph), crtI (crtI -::hph/crtI -::hph) and crtS (crtS -::hph / crtS -::hph) were constructed. All the heterozygote mutants have a pale phenotype and produce less carotenoids than the wild-type strain. The genetic analysis of the crtYB, crtl and crtS loci in the wild-type, heterozygote, and homozygote give evidence of the diploid constitution of ATCC 24230 strains. In addition, the cloning of a truncated form of the crtYB that lacks 153 amino acids of the N-terminal region derived from alternatively spliced mRNA was obtained. Their heterologous expression in Escherichia coli carrying the carotenogenic cluster of Erwinia uredovora result in trans-complementation and give evidence of its functionality in this bacterium, maintaining its phytoene synthase activity but not the lycopene cyclase activity
author	NIKLITSCHEK,MAURICIO ALCAÍNO,JENNIFER BARAHONA,SALVADOR SEPÚLVEDA,DIONISIA LOZANO,CARLA CARMONA,MARISELA MARCOLETA,ANDRÉS MARTÍNEZ,CLAUDIO LODATO,PATRICIA BAEZA,MARCELO CIFUENTES,VÍCTOR
author_facet	NIKLITSCHEK,MAURICIO ALCAÍNO,JENNIFER BARAHONA,SALVADOR SEPÚLVEDA,DIONISIA LOZANO,CARLA CARMONA,MARISELA MARCOLETA,ANDRÉS MARTÍNEZ,CLAUDIO LODATO,PATRICIA BAEZA,MARCELO CIFUENTES,VÍCTOR
author_sort	NIKLITSCHEK,MAURICIO
title	Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous
title_short	Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous
title_full	Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous
title_fullStr	Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous
title_full_unstemmed	Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous
title_sort	genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of xanthophyllomyces dendrorhous
publisher	Sociedad de Biología de Chile
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100011
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Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous

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