Cloning and homologous expression of novel lignin peroxidase genes in the white-rot fungus Phanerochaete sordida YK-624.
Identifieur interne : 000647 ( Main/Exploration ); précédent : 000646; suivant : 000648Cloning and homologous expression of novel lignin peroxidase genes in the white-rot fungus Phanerochaete sordida YK-624.
Auteurs : Tatsuki Sugiura [Japon] ; Kenji Yamagishi ; Toshiyuki Kimura ; Tomoaki Nishida ; Hirokazu Kawagishi ; Hirofumi HiraiSource :
- Bioscience, biotechnology, and biochemistry [ 1347-6947 ] ; 2009.
Descripteurs français
- KwdFr :
- ADN complémentaire (génétique), ADN fongique (MeSH), Clonage moléculaire (MeSH), Données de séquences moléculaires (MeSH), Expression des gènes (MeSH), Génome fongique (génétique), Peroxidases (biosynthèse), Peroxidases (génétique), Phanerochaete (génétique), Plasmides (génétique), Protéines recombinantes (biosynthèse), Protéines recombinantes (génétique), Réaction de polymérisation en chaîne (MeSH), Transformation génétique (MeSH).
- MESH :
- biosynthèse : Peroxidases, Protéines recombinantes.
- génétique : ADN complémentaire, Génome fongique, Peroxidases, Phanerochaete, Plasmides, Protéines recombinantes.
- ADN fongique, Clonage moléculaire, Données de séquences moléculaires, Expression des gènes, Réaction de polymérisation en chaîne, Transformation génétique.
English descriptors
- KwdEn :
- Cloning, Molecular (MeSH), DNA, Complementary (genetics), DNA, Fungal (MeSH), Gene Expression (MeSH), Genome, Fungal (genetics), Molecular Sequence Data (MeSH), Peroxidases (biosynthesis), Peroxidases (genetics), Phanerochaete (genetics), Plasmids (genetics), Polymerase Chain Reaction (MeSH), Recombinant Proteins (biosynthesis), Recombinant Proteins (genetics), Transformation, Genetic (MeSH).
- MESH :
- chemical , biosynthesis : Peroxidases, Recombinant Proteins.
- chemical , genetics : DNA, Complementary, Peroxidases, Recombinant Proteins.
- genetics : Genome, Fungal, Phanerochaete, Plasmids.
- Cloning, Molecular, DNA, Fungal, Gene Expression, Molecular Sequence Data, Polymerase Chain Reaction, Transformation, Genetic.
Abstract
Two genes, encoding YK-LiP1 and YK-LiP2, were cloned from the white-rot fungus Phanerochaete sordida YK-624, and a homologous expression system for the gene was constructed. Two full-length cDNAs (ylpA and ylpB) were isolated by degenerate RT-PCR and RACE-PCR. The results of N-terminal amino acid sequence analysis of native YK-LiP1 and YK-LiP2 showed that ylpA and ylpB coded for YK-LiP2 and YK-LiP1 respectively. The promoter of glyceraldehyde-3-phosphate dehydrogenase cloned from P. sordida YK-624 (PsGPD) was used to drive the expression of ylpA. Expression vector pGPD-g-ylpA was transformed into a P. sordida YK-624 uracil auxotrophic mutant, UV-64. The YlpA protein was secreted in active form by the transformants after 4 d of growth in a medium containing an excessive nitrogen source, whereas endogenous YK-LiP1 and YK-LiP2 were not produced. The physical and catalytic properties of the purified YlpA protein were very similar to those of YK-LiP2. These results suggest that homologous expression of recombinant YK-LiP2 was successful.
DOI: 10.1271/bbb.90152
PubMed: 19661691
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<series><title level="j">Bioscience, biotechnology, and biochemistry</title>
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<term>Gene Expression (MeSH)</term>
<term>Genome, Fungal (genetics)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Peroxidases (biosynthesis)</term>
<term>Peroxidases (genetics)</term>
<term>Phanerochaete (genetics)</term>
<term>Plasmids (genetics)</term>
<term>Polymerase Chain Reaction (MeSH)</term>
<term>Recombinant Proteins (biosynthesis)</term>
<term>Recombinant Proteins (genetics)</term>
<term>Transformation, Genetic (MeSH)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (génétique)</term>
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<term>Clonage moléculaire (MeSH)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Expression des gènes (MeSH)</term>
<term>Génome fongique (génétique)</term>
<term>Peroxidases (biosynthèse)</term>
<term>Peroxidases (génétique)</term>
<term>Phanerochaete (génétique)</term>
<term>Plasmides (génétique)</term>
<term>Protéines recombinantes (biosynthèse)</term>
<term>Protéines recombinantes (génétique)</term>
<term>Réaction de polymérisation en chaîne (MeSH)</term>
<term>Transformation génétique (MeSH)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Peroxidases</term>
<term>Recombinant Proteins</term>
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<term>Phanerochaete</term>
<term>Plasmids</term>
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<term>Peroxidases</term>
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<term>Plasmides</term>
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<term>Gene Expression</term>
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<term>Polymerase Chain Reaction</term>
<term>Transformation, Genetic</term>
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<term>Clonage moléculaire</term>
<term>Données de séquences moléculaires</term>
<term>Expression des gènes</term>
<term>Réaction de polymérisation en chaîne</term>
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<front><div type="abstract" xml:lang="en">Two genes, encoding YK-LiP1 and YK-LiP2, were cloned from the white-rot fungus Phanerochaete sordida YK-624, and a homologous expression system for the gene was constructed. Two full-length cDNAs (ylpA and ylpB) were isolated by degenerate RT-PCR and RACE-PCR. The results of N-terminal amino acid sequence analysis of native YK-LiP1 and YK-LiP2 showed that ylpA and ylpB coded for YK-LiP2 and YK-LiP1 respectively. The promoter of glyceraldehyde-3-phosphate dehydrogenase cloned from P. sordida YK-624 (PsGPD) was used to drive the expression of ylpA. Expression vector pGPD-g-ylpA was transformed into a P. sordida YK-624 uracil auxotrophic mutant, UV-64. The YlpA protein was secreted in active form by the transformants after 4 d of growth in a medium containing an excessive nitrogen source, whereas endogenous YK-LiP1 and YK-LiP2 were not produced. The physical and catalytic properties of the purified YlpA protein were very similar to those of YK-LiP2. These results suggest that homologous expression of recombinant YK-LiP2 was successful.</div>
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<ArticleTitle>Cloning and homologous expression of novel lignin peroxidase genes in the white-rot fungus Phanerochaete sordida YK-624.</ArticleTitle>
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<Abstract><AbstractText>Two genes, encoding YK-LiP1 and YK-LiP2, were cloned from the white-rot fungus Phanerochaete sordida YK-624, and a homologous expression system for the gene was constructed. Two full-length cDNAs (ylpA and ylpB) were isolated by degenerate RT-PCR and RACE-PCR. The results of N-terminal amino acid sequence analysis of native YK-LiP1 and YK-LiP2 showed that ylpA and ylpB coded for YK-LiP2 and YK-LiP1 respectively. The promoter of glyceraldehyde-3-phosphate dehydrogenase cloned from P. sordida YK-624 (PsGPD) was used to drive the expression of ylpA. Expression vector pGPD-g-ylpA was transformed into a P. sordida YK-624 uracil auxotrophic mutant, UV-64. The YlpA protein was secreted in active form by the transformants after 4 d of growth in a medium containing an excessive nitrogen source, whereas endogenous YK-LiP1 and YK-LiP2 were not produced. The physical and catalytic properties of the purified YlpA protein were very similar to those of YK-LiP2. These results suggest that homologous expression of recombinant YK-LiP2 was successful.</AbstractText>
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<country name="Japon"><noRegion><name sortKey="Sugiura, Tatsuki" sort="Sugiura, Tatsuki" uniqKey="Sugiura T" first="Tatsuki" last="Sugiura">Tatsuki Sugiura</name>
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