Production and separation of manganese peroxidase from heme amended yeast cultures.
Identifieur interne : 000679 ( Main/Exploration ); précédent : 000678; suivant : 000680Production and separation of manganese peroxidase from heme amended yeast cultures.
Auteurs : Fei Jiang [États-Unis] ; Puapong Kongsaeree ; Rose Charron ; Curtis Lajoie ; Haowen Xu ; Gary Scott ; Christine KellySource :
- Biotechnology and bioengineering [ 1097-0290 ] ; 2008.
Descripteurs français
- KwdFr :
- Activation enzymatique (MeSH), Hème (métabolisme), Ingénierie des protéines (méthodes), Peroxidases (génétique), Peroxidases (isolement et purification), Peroxidases (métabolisme), Phanerochaete (génétique), Phanerochaete (métabolisme), Pichia (physiologie), Prolifération cellulaire (MeSH), Protéines recombinantes (métabolisme), Stabilité enzymatique (MeSH), Techniques de culture cellulaire (méthodes).
- MESH :
- génétique : Peroxidases, Phanerochaete.
- isolement et purification : Peroxidases.
- métabolisme : Hème, Peroxidases, Phanerochaete, Protéines recombinantes.
- méthodes : Ingénierie des protéines, Techniques de culture cellulaire.
- physiologie : Pichia.
- Activation enzymatique, Prolifération cellulaire, Stabilité enzymatique.
English descriptors
- KwdEn :
- Cell Culture Techniques (methods), Cell Proliferation (MeSH), Enzyme Activation (MeSH), Enzyme Stability (MeSH), Heme (metabolism), Peroxidases (genetics), Peroxidases (isolation & purification), Peroxidases (metabolism), Phanerochaete (genetics), Phanerochaete (metabolism), Pichia (physiology), Protein Engineering (methods), Recombinant Proteins (metabolism).
- MESH :
- chemical , genetics : Peroxidases.
- chemical , isolation & purification : Peroxidases.
- chemical , metabolism : Heme, Peroxidases, Recombinant Proteins.
- genetics : Phanerochaete.
- metabolism : Phanerochaete.
- methods : Cell Culture Techniques, Protein Engineering.
- physiology : Pichia.
- Cell Proliferation, Enzyme Activation, Enzyme Stability.
Abstract
A method for the production and concentration of the lignin-degrading enzyme, manganese peroxidase (rMnP), was developed using the yeast Pichia pastoris in high cell density, fed-batch cultivations. A gene encoding manganese peroxidase (mnp1) from the white-rot fungus Phanerochaete chrysosporium was cloned into a protease deficient (pep4-) strain of the methylotrophic yeast P. pastoris. Heme is an important cofactor for active rMnP production, and amendment of yeast cultures with heme increased active rMnP concentrations. In both shake-flasks and fed-batch bioreactors, the relationship between heme concentration and rMnP activity was logarithmic, with increasing heme concentrations resulting in progressively lesser increases in enzyme activity. Scale-up from shake-flasks to 2 L fed-batch cultivations increased rMnP activities from 200 U/L to 2,500 U/L, with addition of 0.1 g/L heme (added heme per liquid volume) at the beginning of the fed-batch phase resulting in higher enzyme activities than addition at the beginning of the batch phase. A combination of centrifugation, acetone precipitation, dialysis, and freeze drying was found to be effective for concentrating the rMnP from 2,500 U/L in the P. pastoris bioreactor culture to 30,000 U/L in 0.1 M potassium phosphate buffer pH 6. The rMnP recovery yield was 60% and the purity was 1-4%. By using 0.1 g/L heme during the fed-batch cultivation, the heme content of the final enzyme preparation could be reduced by 97%, and had sufficiently high rMnP activity and low enough color to be suitable for pulp bleaching experiments.
DOI: 10.1002/bit.21590
PubMed: 17680655
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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last="Lajoie">Curtis Lajoie</name></author><author><name sortKey="Xu, Haowen" sort="Xu, Haowen" uniqKey="Xu H" first="Haowen" last="Xu">Haowen Xu</name></author><author><name sortKey="Scott, Gary" sort="Scott, Gary" uniqKey="Scott G" first="Gary" last="Scott">Gary Scott</name></author><author><name sortKey="Kelly, Christine" sort="Kelly, Christine" uniqKey="Kelly C" first="Christine" last="Kelly">Christine Kelly</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:17680655</idno><idno type="pmid">17680655</idno><idno type="doi">10.1002/bit.21590</idno><idno type="wicri:Area/Main/Corpus">000726</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000726</idno><idno type="wicri:Area/Main/Curation">000726</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000726</idno><idno 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sort="Lajoie, Curtis" uniqKey="Lajoie C" first="Curtis" last="Lajoie">Curtis Lajoie</name></author><author><name sortKey="Xu, Haowen" sort="Xu, Haowen" uniqKey="Xu H" first="Haowen" last="Xu">Haowen Xu</name></author><author><name sortKey="Scott, Gary" sort="Scott, Gary" uniqKey="Scott G" first="Gary" last="Scott">Gary Scott</name></author><author><name sortKey="Kelly, Christine" sort="Kelly, Christine" uniqKey="Kelly C" first="Christine" last="Kelly">Christine Kelly</name></author></analytic><series><title level="j">Biotechnology and bioengineering</title><idno type="eISSN">1097-0290</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell Culture Techniques (methods)</term><term>Cell Proliferation (MeSH)</term><term>Enzyme Activation (MeSH)</term><term>Enzyme Stability (MeSH)</term><term>Heme (metabolism)</term><term>Peroxidases (genetics)</term><term>Peroxidases (isolation & purification)</term><term>Peroxidases (metabolism)</term><term>Phanerochaete (genetics)</term><term>Phanerochaete (metabolism)</term><term>Pichia (physiology)</term><term>Protein Engineering (methods)</term><term>Recombinant Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Activation enzymatique (MeSH)</term><term>Hème (métabolisme)</term><term>Ingénierie des protéines (méthodes)</term><term>Peroxidases (génétique)</term><term>Peroxidases (isolement et purification)</term><term>Peroxidases (métabolisme)</term><term>Phanerochaete (génétique)</term><term>Phanerochaete (métabolisme)</term><term>Pichia (physiologie)</term><term>Prolifération cellulaire (MeSH)</term><term>Protéines recombinantes (métabolisme)</term><term>Stabilité enzymatique (MeSH)</term><term>Techniques de culture cellulaire (méthodes)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Peroxidases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Peroxidases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Heme</term><term>Peroxidases</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Peroxidases</term><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Peroxidases</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Cell Culture Techniques</term><term>Protein Engineering</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Hème</term><term>Peroxidases</term><term>Phanerochaete</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Ingénierie des protéines</term><term>Techniques de culture cellulaire</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Pichia</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Pichia</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cell Proliferation</term><term>Enzyme Activation</term><term>Enzyme Stability</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Activation enzymatique</term><term>Prolifération cellulaire</term><term>Stabilité enzymatique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A method for the production and concentration of the lignin-degrading enzyme, manganese peroxidase (rMnP), was developed using the yeast Pichia pastoris in high cell density, fed-batch cultivations. A gene encoding manganese peroxidase (mnp1) from the white-rot fungus Phanerochaete chrysosporium was cloned into a protease deficient (pep4-) strain of the methylotrophic yeast P. pastoris. Heme is an important cofactor for active rMnP production, and amendment of yeast cultures with heme increased active rMnP concentrations. In both shake-flasks and fed-batch bioreactors, the relationship between heme concentration and rMnP activity was logarithmic, with increasing heme concentrations resulting in progressively lesser increases in enzyme activity. Scale-up from shake-flasks to 2 L fed-batch cultivations increased rMnP activities from 200 U/L to 2,500 U/L, with addition of 0.1 g/L heme (added heme per liquid volume) at the beginning of the fed-batch phase resulting in higher enzyme activities than addition at the beginning of the batch phase. A combination of centrifugation, acetone precipitation, dialysis, and freeze drying was found to be effective for concentrating the rMnP from 2,500 U/L in the P. pastoris bioreactor culture to 30,000 U/L in 0.1 M potassium phosphate buffer pH 6. The rMnP recovery yield was 60% and the purity was 1-4%. By using 0.1 g/L heme during the fed-batch cultivation, the heme content of the final enzyme preparation could be reduced by 97%, and had sufficiently high rMnP activity and low enough color to be suitable for pulp bleaching experiments.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">17680655</PMID><DateCompleted><Year>2008</Year><Month>01</Month><Day>23</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1097-0290</ISSN><JournalIssue CitedMedium="Internet"><Volume>99</Volume><Issue>3</Issue><PubDate><Year>2008</Year><Month>Feb</Month><Day>15</Day></PubDate></JournalIssue><Title>Biotechnology and bioengineering</Title><ISOAbbreviation>Biotechnol Bioeng</ISOAbbreviation></Journal><ArticleTitle>Production and separation of manganese peroxidase from heme amended yeast cultures.</ArticleTitle><Pagination><MedlinePgn>540-9</MedlinePgn></Pagination><Abstract><AbstractText>A method for the production and concentration of the lignin-degrading enzyme, manganese peroxidase (rMnP), was developed using the yeast Pichia pastoris in high cell density, fed-batch cultivations. A gene encoding manganese peroxidase (mnp1) from the white-rot fungus Phanerochaete chrysosporium was cloned into a protease deficient (pep4-) strain of the methylotrophic yeast P. pastoris. Heme is an important cofactor for active rMnP production, and amendment of yeast cultures with heme increased active rMnP concentrations. In both shake-flasks and fed-batch bioreactors, the relationship between heme concentration and rMnP activity was logarithmic, with increasing heme concentrations resulting in progressively lesser increases in enzyme activity. Scale-up from shake-flasks to 2 L fed-batch cultivations increased rMnP activities from 200 U/L to 2,500 U/L, with addition of 0.1 g/L heme (added heme per liquid volume) at the beginning of the fed-batch phase resulting in higher enzyme activities than addition at the beginning of the batch phase. A combination of centrifugation, acetone precipitation, dialysis, and freeze drying was found to be effective for concentrating the rMnP from 2,500 U/L in the P. pastoris bioreactor culture to 30,000 U/L in 0.1 M potassium phosphate buffer pH 6. The rMnP recovery yield was 60% and the purity was 1-4%. By using 0.1 g/L heme during the fed-batch cultivation, the heme content of the final enzyme preparation could be reduced by 97%, and had sufficiently high rMnP activity and low enough color to be suitable for pulp bleaching experiments.</AbstractText><CopyrightInformation>(c) 2007 Wiley Periodicals, Inc.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Fei</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Cell Genesys, Inc., South San Francisco, California, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kongsaeree</LastName><ForeName>Puapong</ForeName><Initials>P</Initials></Author><Author ValidYN="Y"><LastName>Charron</LastName><ForeName>Rose</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Lajoie</LastName><ForeName>Curtis</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Xu</LastName><ForeName>Haowen</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Scott</LastName><ForeName>Gary</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Kelly</LastName><ForeName>Christine</ForeName><Initials>C</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biotechnol Bioeng</MedlineTA><NlmUniqueID>7502021</NlmUniqueID><ISSNLinking>0006-3592</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>42VZT0U6YR</RegistryNumber><NameOfSubstance UI="D006418">Heme</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.11.1.-</RegistryNumber><NameOfSubstance UI="D010544">Peroxidases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.11.1.13</RegistryNumber><NameOfSubstance UI="C051129">manganese peroxidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D018929" MajorTopicYN="N">Cell Culture Techniques</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D049109" MajorTopicYN="N">Cell Proliferation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004789" MajorTopicYN="N">Enzyme Activation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004795" MajorTopicYN="N">Enzyme Stability</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006418" MajorTopicYN="N">Heme</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010544" MajorTopicYN="N">Peroxidases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020075" MajorTopicYN="N">Phanerochaete</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010843" MajorTopicYN="N">Pichia</DescriptorName><QualifierName UI="Q000502" MajorTopicYN="Y">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015202" MajorTopicYN="N">Protein Engineering</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2007</Year><Month>8</Month><Day>8</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2008</Year><Month>1</Month><Day>24</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2007</Year><Month>8</Month><Day>8</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">17680655</ArticleId><ArticleId IdType="doi">10.1002/bit.21590</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><noCountry><name sortKey="Charron, Rose" sort="Charron, Rose" uniqKey="Charron R" first="Rose" last="Charron">Rose Charron</name><name sortKey="Kelly, Christine" sort="Kelly, Christine" uniqKey="Kelly C" first="Christine" last="Kelly">Christine Kelly</name><name sortKey="Kongsaeree, Puapong" sort="Kongsaeree, Puapong" uniqKey="Kongsaeree P" first="Puapong" last="Kongsaeree">Puapong Kongsaeree</name><name sortKey="Lajoie, Curtis" sort="Lajoie, Curtis" uniqKey="Lajoie C" first="Curtis" last="Lajoie">Curtis Lajoie</name><name sortKey="Scott, Gary" sort="Scott, Gary" uniqKey="Scott G" first="Gary" last="Scott">Gary Scott</name><name sortKey="Xu, Haowen" sort="Xu, Haowen" uniqKey="Xu H" first="Haowen" last="Xu">Haowen Xu</name></noCountry><country name="États-Unis"><region name="Californie"><name sortKey="Jiang, Fei" sort="Jiang, Fei" uniqKey="Jiang F" first="Fei" last="Jiang">Fei Jiang</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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