Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.

Phycomyces carRA gene encodes a protein with two domains. Domain R is characterized by red carR mutants that accumulate lycopene. Domain A is characterized by white carA mutants that do not accumulate significant amounts of carotenoids. The carRA-encoded protein was identified as the lycopene cyclas...

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Autores principales:	Catalina Sanz, Antonio Velayos, María Isabel Álvarez, Ernesto P Benito, Arturo P Eslava
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/284a4ce7474847c1b1b610a108d083d8
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spelling	oai:doaj.org-article:284a4ce7474847c1b1b610a108d083d82021-11-18T06:48:25ZFunctional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.1932-620310.1371/journal.pone.0023102https://doaj.org/article/284a4ce7474847c1b1b610a108d083d82011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21858003/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Phycomyces carRA gene encodes a protein with two domains. Domain R is characterized by red carR mutants that accumulate lycopene. Domain A is characterized by white carA mutants that do not accumulate significant amounts of carotenoids. The carRA-encoded protein was identified as the lycopene cyclase and phytoene synthase enzyme by sequence homology with other proteins. However, no direct data showing the function of this protein have been reported so far. Different Mucor circinelloides mutants altered at the phytoene synthase, the lycopene cyclase or both activities were transformed with the Phycomyces carRA gene. Fully transcribed carRA mRNA molecules were detected by Northern assays in the transformants and the correct processing of the carRA messenger was verified by RT-PCR. These results showed that Phycomyces carRA gene was correctly expressed in Mucor. Carotenoids analysis in these transformants showed the presence of ß-carotene, absent in the untransformed strains, providing functional evidence that the Phycomyces carRA gene complements the M. circinelloides mutations. Co-transformation of the carRA cDNA in E. coli with different combinations of the carotenoid structural genes from Erwinia uredovora was also performed. Newly formed carotenoids were accumulated showing that the Phycomyces CarRA protein does contain lycopene cyclase and phytoene synthase activities. The heterologous expression of the carRA gene and the functional complementation of the mentioned activities are not very efficient in E. coli. However, the simultaneous presence of both carRA and carB gene products from Phycomyces increases the efficiency of these enzymes, presumably due to an interaction mechanism.Catalina SanzAntonio VelayosMaría Isabel ÁlvarezErnesto P BenitoArturo P EslavaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 8, p e23102 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Catalina Sanz Antonio Velayos María Isabel Álvarez Ernesto P Benito Arturo P Eslava Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.
description	Phycomyces carRA gene encodes a protein with two domains. Domain R is characterized by red carR mutants that accumulate lycopene. Domain A is characterized by white carA mutants that do not accumulate significant amounts of carotenoids. The carRA-encoded protein was identified as the lycopene cyclase and phytoene synthase enzyme by sequence homology with other proteins. However, no direct data showing the function of this protein have been reported so far. Different Mucor circinelloides mutants altered at the phytoene synthase, the lycopene cyclase or both activities were transformed with the Phycomyces carRA gene. Fully transcribed carRA mRNA molecules were detected by Northern assays in the transformants and the correct processing of the carRA messenger was verified by RT-PCR. These results showed that Phycomyces carRA gene was correctly expressed in Mucor. Carotenoids analysis in these transformants showed the presence of ß-carotene, absent in the untransformed strains, providing functional evidence that the Phycomyces carRA gene complements the M. circinelloides mutations. Co-transformation of the carRA cDNA in E. coli with different combinations of the carotenoid structural genes from Erwinia uredovora was also performed. Newly formed carotenoids were accumulated showing that the Phycomyces CarRA protein does contain lycopene cyclase and phytoene synthase activities. The heterologous expression of the carRA gene and the functional complementation of the mentioned activities are not very efficient in E. coli. However, the simultaneous presence of both carRA and carB gene products from Phycomyces increases the efficiency of these enzymes, presumably due to an interaction mechanism.
format	article
author	Catalina Sanz Antonio Velayos María Isabel Álvarez Ernesto P Benito Arturo P Eslava
author_facet	Catalina Sanz Antonio Velayos María Isabel Álvarez Ernesto P Benito Arturo P Eslava
author_sort	Catalina Sanz
title	Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.
title_short	Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.
title_full	Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.
title_fullStr	Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.
title_full_unstemmed	Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.
title_sort	functional analysis of the phycomyces carra gene encoding the enzymes phytoene synthase and lycopene cyclase.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/284a4ce7474847c1b1b610a108d083d8
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Functional analysis of the Phycomyces carRA gene encoding the enzymes phytoene synthase and lycopene cyclase.

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