Expression and functional analysis of two lycopene β-cyclases from citrus fruits.
Identifieur interne : 000600 ( PubMed/Checkpoint ); précédent : 000599; suivant : 000601Expression and functional analysis of two lycopene β-cyclases from citrus fruits.
Auteurs : Lancui Zhang [Japon] ; Gang Ma ; Yuki Shirai ; Masaya Kato ; Kazuki Yamawaki ; Yoshinori Ikoma ; Hikaru MatsumotoSource :
- Planta [ 1432-2048 ] ; 2012.
English descriptors
- KwdEn :
- Base Sequence, Biosynthetic Pathways, Carotenoids (analysis), Carotenoids (metabolism), Citrus (enzymology), Citrus (genetics), Escherichia coli (enzymology), Escherichia coli (genetics), Gene Expression, Gene Expression Regulation, Plant (genetics), Genetic Complementation Test, Intramolecular Lyases (genetics), Intramolecular Lyases (metabolism), Molecular Sequence Data, Plant Proteins (genetics), Plant Proteins (metabolism), Recombinant Proteins, Sequence Alignment, Sequence Analysis, DNA, Xanthophylls (analysis), Xanthophylls (metabolism), beta Carotene (analysis), beta Carotene (metabolism).
- MESH :
- chemical , analysis : Carotenoids, Xanthophylls, beta Carotene.
- chemical , genetics : Intramolecular Lyases, Plant Proteins.
- chemical , metabolism : Carotenoids, Intramolecular Lyases, Plant Proteins, Xanthophylls, beta Carotene.
- enzymology : Citrus, Escherichia coli.
- genetics : Citrus, Escherichia coli, Gene Expression Regulation, Plant.
- Base Sequence, Biosynthetic Pathways, Gene Expression, Genetic Complementation Test, Molecular Sequence Data, Recombinant Proteins, Sequence Alignment, Sequence Analysis, DNA.
Abstract
In the present study, two LCYb genes (CitLCYb1 and CitLCYb2) were isolated from Satsuma mandarin (Citrus unshiu Marc.), Valencia orange (Citrus sinensis Osbeck) and Lisbon lemon (Citrus limon Burm.f.) and their functions were analyzed by the color complementation assay in lycopene-accumulating E. coli cells. The results showed that CitLCYb1 and CitLCYb2 shared high identity at the amino acid level among the three citrus varieties. The N-terminal region of the two proteins encoded by CitLCYb1 and CitLCYb2 was predicted to contain a 51-residue chloroplastic transit peptide, which shared low similarity. In Satsuma mandarin, the secondary structures of the CitLCYb1 and CitLCYb2 encoding proteins without the transit peptide were quite similar. Moreover, functional analysis showed that both enzymes of CitLCYb1 and CitLCYb2 participated in the formation of β-carotene, and when they were co-expressed with CitLCYe, α-carotene could be produced from lycopene in E. coli cells. However, although CitLCYb2 could convert lycopene to α-carotene in E. coli cells, its extremely low level of expression indicated that CitLCYb2 did not participate in the formation of α-carotene during the green stage in the flavedo. In addition, the high expression levels of CitLCYb1 and CitLCYb2 during the orange stage played an important role in the accumulation of β,β-xanthophylls in citrus fruits. The results presented in this study might contribute to elucidate the mechanism of carotenoid accumulation in citrus fruits.
DOI: 10.1007/s00425-012-1690-2
PubMed: 22729824
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Expression and functional analysis of two lycopene β-cyclases from citrus fruits.</title><author><name sortKey="Zhang, Lancui" sort="Zhang, Lancui" uniqKey="Zhang L" first="Lancui" last="Zhang">Lancui Zhang</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biological and Environmental Sciences, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Department of Biological and Environmental Sciences, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529</wicri:regionArea><wicri:noRegion>Shizuoka 422-8529</wicri:noRegion></affiliation></author><author><name sortKey="Ma, Gang" sort="Ma, Gang" uniqKey="Ma G" first="Gang" last="Ma">Gang Ma</name></author><author><name sortKey="Shirai, Yuki" sort="Shirai, Yuki" uniqKey="Shirai Y" first="Yuki" last="Shirai">Yuki Shirai</name></author><author><name sortKey="Kato, Masaya" sort="Kato, Masaya" uniqKey="Kato M" first="Masaya" last="Kato">Masaya Kato</name></author><author><name sortKey="Yamawaki, Kazuki" sort="Yamawaki, Kazuki" uniqKey="Yamawaki K" first="Kazuki" last="Yamawaki">Kazuki Yamawaki</name></author><author><name sortKey="Ikoma, Yoshinori" sort="Ikoma, Yoshinori" uniqKey="Ikoma Y" first="Yoshinori" last="Ikoma">Yoshinori Ikoma</name></author><author><name sortKey="Matsumoto, Hikaru" sort="Matsumoto, Hikaru" uniqKey="Matsumoto H" first="Hikaru" last="Matsumoto">Hikaru Matsumoto</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2012">2012</date><idno type="RBID">pubmed:22729824</idno><idno type="pmid">22729824</idno><idno type="doi">10.1007/s00425-012-1690-2</idno><idno type="wicri:Area/PubMed/Corpus">000568</idno><idno type="wicri:Area/PubMed/Curation">000568</idno><idno 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uniqKey="Shirai Y" first="Yuki" last="Shirai">Yuki Shirai</name></author><author><name sortKey="Kato, Masaya" sort="Kato, Masaya" uniqKey="Kato M" first="Masaya" last="Kato">Masaya Kato</name></author><author><name sortKey="Yamawaki, Kazuki" sort="Yamawaki, Kazuki" uniqKey="Yamawaki K" first="Kazuki" last="Yamawaki">Kazuki Yamawaki</name></author><author><name sortKey="Ikoma, Yoshinori" sort="Ikoma, Yoshinori" uniqKey="Ikoma Y" first="Yoshinori" last="Ikoma">Yoshinori Ikoma</name></author><author><name sortKey="Matsumoto, Hikaru" sort="Matsumoto, Hikaru" uniqKey="Matsumoto H" first="Hikaru" last="Matsumoto">Hikaru Matsumoto</name></author></analytic><series><title level="j">Planta</title><idno type="eISSN">1432-2048</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term><term>Biosynthetic Pathways</term><term>Carotenoids (analysis)</term><term>Carotenoids (metabolism)</term><term>Citrus (enzymology)</term><term>Citrus (genetics)</term><term>Escherichia coli (enzymology)</term><term>Escherichia coli (genetics)</term><term>Gene Expression</term><term>Gene Expression Regulation, Plant (genetics)</term><term>Genetic Complementation Test</term><term>Intramolecular Lyases (genetics)</term><term>Intramolecular Lyases (metabolism)</term><term>Molecular Sequence Data</term><term>Plant Proteins (genetics)</term><term>Plant Proteins (metabolism)</term><term>Recombinant Proteins</term><term>Sequence Alignment</term><term>Sequence Analysis, DNA</term><term>Xanthophylls (analysis)</term><term>Xanthophylls (metabolism)</term><term>beta Carotene (analysis)</term><term>beta Carotene (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Carotenoids</term><term>Xanthophylls</term><term>beta Carotene</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Intramolecular Lyases</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Carotenoids</term><term>Intramolecular Lyases</term><term>Plant Proteins</term><term>Xanthophylls</term><term>beta Carotene</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Citrus</term><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Citrus</term><term>Escherichia coli</term><term>Gene Expression Regulation, Plant</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Biosynthetic Pathways</term><term>Gene Expression</term><term>Genetic Complementation Test</term><term>Molecular Sequence Data</term><term>Recombinant Proteins</term><term>Sequence Alignment</term><term>Sequence Analysis, DNA</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In the present study, two LCYb genes (CitLCYb1 and CitLCYb2) were isolated from Satsuma mandarin (Citrus unshiu Marc.), Valencia orange (Citrus sinensis Osbeck) and Lisbon lemon (Citrus limon Burm.f.) and their functions were analyzed by the color complementation assay in lycopene-accumulating E. coli cells. The results showed that CitLCYb1 and CitLCYb2 shared high identity at the amino acid level among the three citrus varieties. The N-terminal region of the two proteins encoded by CitLCYb1 and CitLCYb2 was predicted to contain a 51-residue chloroplastic transit peptide, which shared low similarity. In Satsuma mandarin, the secondary structures of the CitLCYb1 and CitLCYb2 encoding proteins without the transit peptide were quite similar. Moreover, functional analysis showed that both enzymes of CitLCYb1 and CitLCYb2 participated in the formation of β-carotene, and when they were co-expressed with CitLCYe, α-carotene could be produced from lycopene in E. coli cells. However, although CitLCYb2 could convert lycopene to α-carotene in E. coli cells, its extremely low level of expression indicated that CitLCYb2 did not participate in the formation of α-carotene during the green stage in the flavedo. In addition, the high expression levels of CitLCYb1 and CitLCYb2 during the orange stage played an important role in the accumulation of β,β-xanthophylls in citrus fruits. The results presented in this study might contribute to elucidate the mechanism of carotenoid accumulation in citrus fruits.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22729824</PMID><DateCreated><Year>2012</Year><Month>9</Month><Day>27</Day></DateCreated><DateCompleted><Year>2013</Year><Month>09</Month><Day>05</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1432-2048</ISSN><JournalIssue CitedMedium="Internet"><Volume>236</Volume><Issue>4</Issue><PubDate><Year>2012</Year><Month>Oct</Month></PubDate></JournalIssue><Title>Planta</Title><ISOAbbreviation>Planta</ISOAbbreviation></Journal><ArticleTitle>Expression and functional analysis of two lycopene β-cyclases from citrus fruits.</ArticleTitle><Pagination><MedlinePgn>1315-25</MedlinePgn></Pagination><Abstract><AbstractText>In the present study, two LCYb genes (CitLCYb1 and CitLCYb2) were isolated from Satsuma mandarin (Citrus unshiu Marc.), Valencia orange (Citrus sinensis Osbeck) and Lisbon lemon (Citrus limon Burm.f.) and their functions were analyzed by the color complementation assay in lycopene-accumulating E. coli cells. The results showed that CitLCYb1 and CitLCYb2 shared high identity at the amino acid level among the three citrus varieties. The N-terminal region of the two proteins encoded by CitLCYb1 and CitLCYb2 was predicted to contain a 51-residue chloroplastic transit peptide, which shared low similarity. In Satsuma mandarin, the secondary structures of the CitLCYb1 and CitLCYb2 encoding proteins without the transit peptide were quite similar. Moreover, functional analysis showed that both enzymes of CitLCYb1 and CitLCYb2 participated in the formation of β-carotene, and when they were co-expressed with CitLCYe, α-carotene could be produced from lycopene in E. coli cells. However, although CitLCYb2 could convert lycopene to α-carotene in E. coli cells, its extremely low level of expression indicated that CitLCYb2 did not participate in the formation of α-carotene during the green stage in the flavedo. In addition, the high expression levels of CitLCYb1 and CitLCYb2 during the orange stage played an important role in the accumulation of β,β-xanthophylls in citrus fruits. The results presented in this study might contribute to elucidate the mechanism of carotenoid accumulation in citrus fruits.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Lancui</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Department of Biological and Environmental Sciences, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ma</LastName><ForeName>Gang</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Shirai</LastName><ForeName>Yuki</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Kato</LastName><ForeName>Masaya</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Yamawaki</LastName><ForeName>Kazuki</ForeName><Initials>K</Initials></Author><Author ValidYN="Y"><LastName>Ikoma</LastName><ForeName>Yoshinori</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Matsumoto</LastName><ForeName>Hikaru</ForeName><Initials>H</Initials></Author></AuthorList><Language>ENG</Language><DataBankList CompleteYN="Y"><DataBank><DataBankName>GENBANK</DataBankName><AccessionNumberList><AccessionNumber>AB114652</AccessionNumber><AccessionNumber>AB114660</AccessionNumber><AccessionNumber>AB114668</AccessionNumber><AccessionNumber>AB719392</AccessionNumber><AccessionNumber>AB719393</AccessionNumber><AccessionNumber>AB719394</AccessionNumber></AccessionNumberList></DataBank></DataBankList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2012</Year><Month>Jun</Month><Day>24</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Planta</MedlineTA><NlmUniqueID>1250576</NlmUniqueID><ISSNLinking>0032-0935</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D024341">Xanthophylls</NameOfSubstance></Chemical><Chemical><RegistryNumber>01YAE03M7J</RegistryNumber><NameOfSubstance UI="D019207">beta Carotene</NameOfSubstance></Chemical><Chemical><RegistryNumber>36-88-4</RegistryNumber><NameOfSubstance UI="D002338">Carotenoids</NameOfSubstance></Chemical><Chemical><RegistryNumber>45XWE1Z69V</RegistryNumber><NameOfSubstance 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MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010940" MajorTopicYN="N">Plant Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016415" MajorTopicYN="N">Sequence Alignment</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D017422" MajorTopicYN="N">Sequence Analysis, DNA</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D024341" MajorTopicYN="N">Xanthophylls</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000378" 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PubStatus="medline"><Year>2013</Year><Month>9</Month><Day>6</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">22729824</ArticleId><ArticleId IdType="doi">10.1007/s00425-012-1690-2</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Japon</li></country></list><tree><noCountry><name sortKey="Ikoma, Yoshinori" sort="Ikoma, Yoshinori" uniqKey="Ikoma Y" first="Yoshinori" last="Ikoma">Yoshinori Ikoma</name><name sortKey="Kato, Masaya" sort="Kato, Masaya" uniqKey="Kato M" first="Masaya" last="Kato">Masaya Kato</name><name sortKey="Ma, Gang" sort="Ma, Gang" uniqKey="Ma G" first="Gang" last="Ma">Gang Ma</name><name sortKey="Matsumoto, Hikaru" sort="Matsumoto, Hikaru" uniqKey="Matsumoto H" first="Hikaru" last="Matsumoto">Hikaru Matsumoto</name><name sortKey="Shirai, Yuki" sort="Shirai, Yuki" uniqKey="Shirai Y" first="Yuki" last="Shirai">Yuki Shirai</name><name sortKey="Yamawaki, Kazuki" sort="Yamawaki, Kazuki" uniqKey="Yamawaki K" first="Kazuki" last="Yamawaki">Kazuki Yamawaki</name></noCountry><country name="Japon"><noRegion><name sortKey="Zhang, Lancui" sort="Zhang, Lancui" uniqKey="Zhang L" first="Lancui" last="Zhang">Lancui Zhang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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