Production of poplar xyloglucan endotransglycosylase using the methylotrophic yeast Pichia pastoris.
Identifieur interne : 003F71 ( Main/Exploration ); précédent : 003F70; suivant : 003F72Production of poplar xyloglucan endotransglycosylase using the methylotrophic yeast Pichia pastoris.
Auteurs : Monika Bollok [Suède] ; Hongbin Henriksson ; Asa Kallas ; Mehmedalija Jahic ; Tuula T. Teeri ; Sven-Olof EnforsSource :
- Applied biochemistry and biotechnology [ 0273-2289 ] ; 2005.
Descripteurs français
- KwdFr :
- Activation enzymatique (MeSH), Glycosyltransferase (biosynthèse), Glycosyltransferase (génétique), Ingénierie des protéines (méthodes), Méthanol (métabolisme), Pichia (croissance et développement), Pichia (enzymologie), Pichia (génétique), Populus (enzymologie), Populus (génétique), Protéines recombinantes (biosynthèse), Techniques de culture cellulaire (méthodes).
- MESH :
- biosynthèse : Glycosyltransferase, Protéines recombinantes.
- croissance et développement : Pichia.
- enzymologie : Pichia, Populus.
- génétique : Glycosyltransferase, Pichia, Populus.
- métabolisme : Méthanol.
- méthodes : Ingénierie des protéines, Techniques de culture cellulaire.
- Activation enzymatique.
English descriptors
- KwdEn :
- Cell Culture Techniques (methods), Enzyme Activation (MeSH), Glycosyltransferases (biosynthesis), Glycosyltransferases (genetics), Methanol (metabolism), Pichia (enzymology), Pichia (genetics), Pichia (growth & development), Populus (enzymology), Populus (genetics), Protein Engineering (methods), Recombinant Proteins (biosynthesis).
- MESH :
- chemical , biosynthesis : Glycosyltransferases, Recombinant Proteins.
- chemical , genetics : Glycosyltransferases.
- chemical , metabolism : Methanol.
- enzymology : Pichia, Populus.
- genetics : Pichia, Populus.
- growth & development : Pichia.
- methods : Cell Culture Techniques, Protein Engineering.
- Enzyme Activation.
Abstract
The gene XET16A encoding the enzyme xyloglucan endotransglycosylase (XET) from hybrid aspen (Populus tremula x tremuloides Mich) was transformed into Pichia pastoris GS115 and the enzyme was secreted to the medium. The influence of process conditions on the XET production, activity, and proteolytic degradation were examined. Inactivation of XET occurred in the foam, but could be decreased significantly by using an efficient antifoam. Rich medium (yeast extract plus peptone) was needed for product accumulation, but not for growth. The proteolytic degradation of the enzyme in the medium was substantially decreased by also adding yeast extract and peptone to the glycerol medium before induction with methanol. Decreasing the fermentation pH from 5.0 to 4.0 further reduced the proteolysis. The specific activity was further improved by production at 15 degrees C instead of 22 degrees C. In this way a XET production of 54 mg/L active enzyme could be achieved in the process with a specific activity of 18 Unit/mg protein after a downstream process including centrifugation, micro- and ultrafiltration, and ion exchange chromatography.
DOI: 10.1007/s12010-005-0006-4
PubMed: 16014999
Affiliations:
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Teeri</name></author><author><name sortKey="Enfors, Sven Olof" sort="Enfors, Sven Olof" uniqKey="Enfors S" first="Sven-Olof" last="Enfors">Sven-Olof Enfors</name></author></analytic><series><title level="j">Applied biochemistry and biotechnology</title><idno type="ISSN">0273-2289</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell Culture Techniques (methods)</term><term>Enzyme Activation (MeSH)</term><term>Glycosyltransferases (biosynthesis)</term><term>Glycosyltransferases (genetics)</term><term>Methanol (metabolism)</term><term>Pichia (enzymology)</term><term>Pichia (genetics)</term><term>Pichia (growth & development)</term><term>Populus (enzymology)</term><term>Populus (genetics)</term><term>Protein Engineering (methods)</term><term>Recombinant Proteins (biosynthesis)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Activation enzymatique (MeSH)</term><term>Glycosyltransferase (biosynthèse)</term><term>Glycosyltransferase (génétique)</term><term>Ingénierie des protéines (méthodes)</term><term>Méthanol (métabolisme)</term><term>Pichia (croissance et développement)</term><term>Pichia (enzymologie)</term><term>Pichia (génétique)</term><term>Populus (enzymologie)</term><term>Populus (génétique)</term><term>Protéines recombinantes (biosynthèse)</term><term>Techniques de culture cellulaire (méthodes)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Glycosyltransferases</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Glycosyltransferases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Methanol</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Glycosyltransferase</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Pichia</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Pichia</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Pichia</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Pichia</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Pichia</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycosyltransferase</term><term>Pichia</term><term>Populus</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>Méthanol</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" xml:lang="en"><term>Enzyme Activation</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Activation enzymatique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The gene XET16A encoding the enzyme xyloglucan endotransglycosylase (XET) from hybrid aspen (Populus tremula x tremuloides Mich) was transformed into Pichia pastoris GS115 and the enzyme was secreted to the medium. The influence of process conditions on the XET production, activity, and proteolytic degradation were examined. Inactivation of XET occurred in the foam, but could be decreased significantly by using an efficient antifoam. Rich medium (yeast extract plus peptone) was needed for product accumulation, but not for growth. The proteolytic degradation of the enzyme in the medium was substantially decreased by also adding yeast extract and peptone to the glycerol medium before induction with methanol. Decreasing the fermentation pH from 5.0 to 4.0 further reduced the proteolysis. The specific activity was further improved by production at 15 degrees C instead of 22 degrees C. In this way a XET production of 54 mg/L active enzyme could be achieved in the process with a specific activity of 18 Unit/mg protein after a downstream process including centrifugation, micro- and ultrafiltration, and ion exchange chromatography.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16014999</PMID><DateCompleted><Year>2005</Year><Month>10</Month><Day>19</Day></DateCompleted><DateRevised><Year>2019</Year><Month>09</Month><Day>11</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0273-2289</ISSN><JournalIssue CitedMedium="Print"><Volume>126</Volume><Issue>1</Issue><PubDate><Year>2005</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Applied biochemistry and biotechnology</Title><ISOAbbreviation>Appl Biochem Biotechnol</ISOAbbreviation></Journal><ArticleTitle>Production of poplar xyloglucan endotransglycosylase using the methylotrophic yeast Pichia pastoris.</ArticleTitle><Pagination><MedlinePgn>61-77</MedlinePgn></Pagination><Abstract><AbstractText>The gene XET16A encoding the enzyme xyloglucan endotransglycosylase (XET) from hybrid aspen (Populus tremula x tremuloides Mich) was transformed into Pichia pastoris GS115 and the enzyme was secreted to the medium. The influence of process conditions on the XET production, activity, and proteolytic degradation were examined. Inactivation of XET occurred in the foam, but could be decreased significantly by using an efficient antifoam. Rich medium (yeast extract plus peptone) was needed for product accumulation, but not for growth. The proteolytic degradation of the enzyme in the medium was substantially decreased by also adding yeast extract and peptone to the glycerol medium before induction with methanol. Decreasing the fermentation pH from 5.0 to 4.0 further reduced the proteolysis. The specific activity was further improved by production at 15 degrees C instead of 22 degrees C. In this way a XET production of 54 mg/L active enzyme could be achieved in the process with a specific activity of 18 Unit/mg protein after a downstream process including centrifugation, micro- and ultrafiltration, and ion exchange chromatography.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Bollok</LastName><ForeName>Monika</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Biotechnology, Royal Institute of Technology, KTH, Alba Nova University Center, Roslagstullsbacken 21, S-106 91, Stockholm, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Henriksson</LastName><ForeName>Hongbin</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Kallas</LastName><ForeName>Asa</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Jahic</LastName><ForeName>Mehmedalija</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Teeri</LastName><ForeName>Tuula T</ForeName><Initials>TT</Initials></Author><Author ValidYN="Y"><LastName>Enfors</LastName><ForeName>Sven-Olof</ForeName><Initials>SO</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Appl Biochem Biotechnol</MedlineTA><NlmUniqueID>8208561</NlmUniqueID><ISSNLinking>0273-2289</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.4.-</RegistryNumber><NameOfSubstance UI="D016695">Glycosyltransferases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.4.1.207</RegistryNumber><NameOfSubstance UI="C473049">xyloglucan - xyloglucosyltransferase</NameOfSubstance></Chemical><Chemical><RegistryNumber>Y4S76JWI15</RegistryNumber><NameOfSubstance UI="D000432">Methanol</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="D004789" MajorTopicYN="N">Enzyme Activation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016695" MajorTopicYN="N">Glycosyltransferases</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000432" MajorTopicYN="N">Methanol</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010843" MajorTopicYN="N">Pichia</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D032107" MajorTopicYN="N">Populus</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</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="Q000096" MajorTopicYN="N">biosynthesis</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2004</Year><Month>07</Month><Day>09</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2005</Year><Month>03</Month><Day>22</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2005</Year><Month>7</Month><Day>15</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2005</Year><Month>10</Month><Day>20</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2005</Year><Month>7</Month><Day>15</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">16014999</ArticleId><ArticleId IdType="pii">ABAB:126:1:061</ArticleId><ArticleId IdType="doi">10.1007/s12010-005-0006-4</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Suède</li></country><region><li>Svealand</li></region><settlement><li>Stockholm</li></settlement></list><tree><noCountry><name sortKey="Enfors, Sven Olof" sort="Enfors, Sven Olof" uniqKey="Enfors S" first="Sven-Olof" last="Enfors">Sven-Olof Enfors</name><name sortKey="Henriksson, Hongbin" sort="Henriksson, Hongbin" uniqKey="Henriksson H" first="Hongbin" last="Henriksson">Hongbin Henriksson</name><name sortKey="Jahic, Mehmedalija" sort="Jahic, Mehmedalija" uniqKey="Jahic M" first="Mehmedalija" last="Jahic">Mehmedalija Jahic</name><name sortKey="Kallas, Asa" sort="Kallas, Asa" uniqKey="Kallas A" first="Asa" last="Kallas">Asa Kallas</name><name sortKey="Teeri, Tuula T" sort="Teeri, Tuula T" uniqKey="Teeri T" first="Tuula T" last="Teeri">Tuula T. Teeri</name></noCountry><country name="Suède"><region name="Svealand"><name sortKey="Bollok, Monika" sort="Bollok, Monika" uniqKey="Bollok M" first="Monika" last="Bollok">Monika Bollok</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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