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...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2008-01-01
|
| Series: | Biological Research |
| Subjects: | |
| Online Access: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100011 |
| _version_ | 1819178852636688384 |
|---|---|
| author | MAURICIO NIKLITSCHEK JENNIFER ALCAÍNO SALVADOR BARAHONA DIONISIA SEPÚLVEDA CARLA LOZANO MARISELA CARMONA ANDRÉS MARCOLETA CLAUDIO MARTÍNEZ PATRICIA LODATO MARCELO BAEZA VÍCTOR CIFUENTES |
| author_facet | MAURICIO NIKLITSCHEK JENNIFER ALCAÍNO SALVADOR BARAHONA DIONISIA SEPÚLVEDA CARLA LOZANO MARISELA CARMONA ANDRÉS MARCOLETA CLAUDIO MARTÍNEZ PATRICIA LODATO MARCELO BAEZA VÍCTOR CIFUENTES |
| author_sort | MAURICIO NIKLITSCHEK |
| collection | DOAJ |
| 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 |
| first_indexed | 2024-12-22T21:49:08Z |
| format | Article |
| id | doaj.art-8b10acf404974bf78161d4fe13cf5476 |
| institution | Directory Open Access Journal |
| issn | 0716-9760 0717-6287 |
| language | English |
| last_indexed | 2024-12-22T21:49:08Z |
| publishDate | 2008-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biological Research |
| spelling | doaj.art-8b10acf404974bf78161d4fe13cf54762022-12-21T18:11:25ZengBMCBiological Research0716-97600717-62872008-01-0141193108Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhousMAURICIO NIKLITSCHEKJENNIFER ALCAÍNOSALVADOR BARAHONADIONISIA SEPÚLVEDACARLA LOZANOMARISELA CARMONAANDRÉS MARCOLETACLAUDIO MARTÍNEZPATRICIA LODATOMARCELO BAEZAVÍCTOR CIFUENTESThe 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 activityhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100011astaxanthin biosynthesisXanthophyllomyces dendrorhousPhaffia rhodozyma |
| spellingShingle | MAURICIO NIKLITSCHEK JENNIFER ALCAÍNO SALVADOR BARAHONA DIONISIA SEPÚLVEDA CARLA LOZANO MARISELA CARMONA ANDRÉS MARCOLETA CLAUDIO MARTÍNEZ PATRICIA LODATO MARCELO BAEZA VÍCTOR CIFUENTES Genomic organization of the structural genes controlling the astaxanthin biosynthesis pathway of Xanthophyllomyces dendrorhous Biological Research astaxanthin biosynthesis Xanthophyllomyces dendrorhous Phaffia rhodozyma |
| title | 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_short | 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 |
| topic | astaxanthin biosynthesis Xanthophyllomyces dendrorhous Phaffia rhodozyma |
| url | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100011 |
| work_keys_str_mv | AT mauricioniklitschek genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT jenniferalcaino genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT salvadorbarahona genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT dionisiasepulveda genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT carlalozano genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT mariselacarmona genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT andresmarcoleta genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT claudiomartinez genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT patricialodato genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT marcelobaeza genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous AT victorcifuentes genomicorganizationofthestructuralgenescontrollingtheastaxanthinbiosynthesispathwayofxanthophyllomycesdendrorhous |