Metabolic engineering of glycerol production in Saccharomyces cerevisiae.
Identifieur interne : 002761 ( Main/Exploration ); précédent : 002760; suivant : 002762Metabolic engineering of glycerol production in Saccharomyces cerevisiae.
Auteurs : Karin M. Overkamp [Pays-Bas] ; Barbara M. Bakker ; Peter Kötter ; Marijke A H. Luttik ; Johannes P. Van Dijken ; Jack T. PronkSource :
- Applied and environmental microbiology [ 0099-2240 ] ; 2002.
Descripteurs français
- KwdFr :
- Cytosol (métabolisme), Glucose (métabolisme), Glycérol (métabolisme), Glycérophosphate (métabolisme), Génie génétique (MeSH), Mitochondries (métabolisme), NAD (métabolisme), Oxydoréduction (MeSH), Phénotype (MeSH), Saccharomyces cerevisiae (croissance et développement), Saccharomyces cerevisiae (métabolisme), Techniques de culture cellulaire (MeSH).
- MESH :
- croissance et développement : Saccharomyces cerevisiae.
- métabolisme : Cytosol, Glucose, Glycérol, Glycérophosphate, Mitochondries, NAD, Saccharomyces cerevisiae.
- Génie génétique, Oxydoréduction, Phénotype, Techniques de culture cellulaire.
English descriptors
- KwdEn :
- Cell Culture Techniques (MeSH), Cytosol (metabolism), Genetic Engineering (MeSH), Glucose (metabolism), Glycerol (metabolism), Glycerophosphates (metabolism), Mitochondria (metabolism), NAD (metabolism), Oxidation-Reduction (MeSH), Phenotype (MeSH), Saccharomyces cerevisiae (growth & development), Saccharomyces cerevisiae (metabolism).
- MESH :
- chemical , metabolism : Glucose, Glycerol, Glycerophosphates, NAD.
- growth & development : Saccharomyces cerevisiae.
- metabolism : Cytosol, Mitochondria, Saccharomyces cerevisiae.
- Cell Culture Techniques, Genetic Engineering, Oxidation-Reduction, Phenotype.
Abstract
Inactivation of TPI1, the Saccharomyces cerevisiae structural gene encoding triose phosphate isomerase, completely eliminates growth on glucose as the sole carbon source. In tpi1-null mutants, intracellular accumulation of dihydroxyacetone phosphate might be prevented if the cytosolic NADH generated in glycolysis by glyceraldehyde-3-phosphate dehydrogenase were quantitatively used to reduce dihydroxyacetone phosphate to glycerol. We hypothesize that the growth defect of tpi1-null mutants is caused by mitochondrial reoxidation of cytosolic NADH, thus rendering it unavailable for dihydroxyacetone-phosphate reduction. To test this hypothesis, a tpi1delta nde1delta nde2delta gut2delta quadruple mutant was constructed. NDE1 and NDE2 encode isoenzymes of mitochondrial external NADH dehydrogenase; GUT2 encodes a key enzyme of the glycerol-3-phosphate shuttle. It has recently been demonstrated that these two systems are primarily responsible for mitochondrial oxidation of cytosolic NADH in S. cerevisiae. Consistent with the hypothesis, the quadruple mutant grew on glucose as the sole carbon source. The growth on glucose, which was accompanied by glycerol production, was inhibited at high-glucose concentrations. This inhibition was attributed to glucose repression of respiratory enzymes as, in the quadruple mutant, respiratory pyruvate dissimilation is essential for ATP synthesis and growth. Serial transfer of the quadruple mutant on high-glucose media yielded a spontaneous mutant with much higher specific growth rates in high-glucose media (up to 0.10 h(-1) at 100 g of glucose. liter(-1)). In aerated batch cultures grown on 400 g of glucose. liter(-1), this engineered S. cerevisiae strain produced over 200 g of glycerol. liter(-1), corresponding to a molar yield of glycerol on glucose close to unity.
DOI: 10.1128/aem.68.6.2814-2821.2002
PubMed: 12039737
PubMed Central: PMC123913
Affiliations:
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Overkamp</name><affiliation wicri:level="3"><nlm:affiliation>Kluyver Laboratory of Biotechnology, Delft University of Technology, NL-2628 BC Delft, Amsterdam.</nlm:affiliation><country>Pays-Bas</country><placeName><settlement type="city">Amsterdam</settlement><region nuts="2" type="province">Hollande-Septentrionale</region></placeName><wicri:orgArea>Kluyver Laboratory of Biotechnology, Delft University of Technology, NL-2628 BC Delft</wicri:orgArea></affiliation></author><author><name sortKey="Bakker, Barbara M" sort="Bakker, Barbara M" uniqKey="Bakker B" first="Barbara M" last="Bakker">Barbara M. Bakker</name></author><author><name sortKey="Kotter, Peter" sort="Kotter, Peter" uniqKey="Kotter P" first="Peter" last="Kötter">Peter Kötter</name></author><author><name sortKey="Luttik, Marijke A H" sort="Luttik, Marijke A H" uniqKey="Luttik M" first="Marijke A H" last="Luttik">Marijke A H. Luttik</name></author><author><name sortKey="Van Dijken, Johannes P" sort="Van Dijken, Johannes P" uniqKey="Van Dijken J" first="Johannes P" last="Van Dijken">Johannes P. Van Dijken</name></author><author><name sortKey="Pronk, Jack T" sort="Pronk, Jack T" uniqKey="Pronk J" first="Jack T" last="Pronk">Jack T. Pronk</name></author></analytic><series><title level="j">Applied and environmental microbiology</title><idno type="ISSN">0099-2240</idno><imprint><date when="2002" type="published">2002</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell Culture Techniques (MeSH)</term><term>Cytosol (metabolism)</term><term>Genetic Engineering (MeSH)</term><term>Glucose (metabolism)</term><term>Glycerol (metabolism)</term><term>Glycerophosphates (metabolism)</term><term>Mitochondria (metabolism)</term><term>NAD (metabolism)</term><term>Oxidation-Reduction (MeSH)</term><term>Phenotype (MeSH)</term><term>Saccharomyces cerevisiae (growth & development)</term><term>Saccharomyces cerevisiae (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Cytosol (métabolisme)</term><term>Glucose (métabolisme)</term><term>Glycérol (métabolisme)</term><term>Glycérophosphate (métabolisme)</term><term>Génie génétique (MeSH)</term><term>Mitochondries (métabolisme)</term><term>NAD (métabolisme)</term><term>Oxydoréduction (MeSH)</term><term>Phénotype (MeSH)</term><term>Saccharomyces cerevisiae (croissance et développement)</term><term>Saccharomyces cerevisiae (métabolisme)</term><term>Techniques de culture cellulaire (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glucose</term><term>Glycerol</term><term>Glycerophosphates</term><term>NAD</term></keywords><keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Saccharomyces cerevisiae</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Saccharomyces cerevisiae</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Cytosol</term><term>Mitochondria</term><term>Saccharomyces cerevisiae</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cytosol</term><term>Glucose</term><term>Glycérol</term><term>Glycérophosphate</term><term>Mitochondries</term><term>NAD</term><term>Saccharomyces cerevisiae</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cell Culture Techniques</term><term>Genetic Engineering</term><term>Oxidation-Reduction</term><term>Phenotype</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Génie génétique</term><term>Oxydoréduction</term><term>Phénotype</term><term>Techniques de culture cellulaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Inactivation of TPI1, the Saccharomyces cerevisiae structural gene encoding triose phosphate isomerase, completely eliminates growth on glucose as the sole carbon source. In tpi1-null mutants, intracellular accumulation of dihydroxyacetone phosphate might be prevented if the cytosolic NADH generated in glycolysis by glyceraldehyde-3-phosphate dehydrogenase were quantitatively used to reduce dihydroxyacetone phosphate to glycerol. We hypothesize that the growth defect of tpi1-null mutants is caused by mitochondrial reoxidation of cytosolic NADH, thus rendering it unavailable for dihydroxyacetone-phosphate reduction. To test this hypothesis, a tpi1delta nde1delta nde2delta gut2delta quadruple mutant was constructed. NDE1 and NDE2 encode isoenzymes of mitochondrial external NADH dehydrogenase; GUT2 encodes a key enzyme of the glycerol-3-phosphate shuttle. It has recently been demonstrated that these two systems are primarily responsible for mitochondrial oxidation of cytosolic NADH in S. cerevisiae. Consistent with the hypothesis, the quadruple mutant grew on glucose as the sole carbon source. The growth on glucose, which was accompanied by glycerol production, was inhibited at high-glucose concentrations. This inhibition was attributed to glucose repression of respiratory enzymes as, in the quadruple mutant, respiratory pyruvate dissimilation is essential for ATP synthesis and growth. Serial transfer of the quadruple mutant on high-glucose media yielded a spontaneous mutant with much higher specific growth rates in high-glucose media (up to 0.10 h(-1) at 100 g of glucose. liter(-1)). In aerated batch cultures grown on 400 g of glucose. liter(-1), this engineered S. cerevisiae strain produced over 200 g of glycerol. liter(-1), corresponding to a molar yield of glycerol on glucose close to unity.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">12039737</PMID><DateCompleted><Year>2002</Year><Month>07</Month><Day>22</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>08</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0099-2240</ISSN><JournalIssue CitedMedium="Print"><Volume>68</Volume><Issue>6</Issue><PubDate><Year>2002</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Applied and environmental microbiology</Title><ISOAbbreviation>Appl Environ Microbiol</ISOAbbreviation></Journal><ArticleTitle>Metabolic engineering of glycerol production in Saccharomyces cerevisiae.</ArticleTitle><Pagination><MedlinePgn>2814-21</MedlinePgn></Pagination><Abstract><AbstractText>Inactivation of TPI1, the Saccharomyces cerevisiae structural gene encoding triose phosphate isomerase, completely eliminates growth on glucose as the sole carbon source. In tpi1-null mutants, intracellular accumulation of dihydroxyacetone phosphate might be prevented if the cytosolic NADH generated in glycolysis by glyceraldehyde-3-phosphate dehydrogenase were quantitatively used to reduce dihydroxyacetone phosphate to glycerol. We hypothesize that the growth defect of tpi1-null mutants is caused by mitochondrial reoxidation of cytosolic NADH, thus rendering it unavailable for dihydroxyacetone-phosphate reduction. To test this hypothesis, a tpi1delta nde1delta nde2delta gut2delta quadruple mutant was constructed. NDE1 and NDE2 encode isoenzymes of mitochondrial external NADH dehydrogenase; GUT2 encodes a key enzyme of the glycerol-3-phosphate shuttle. It has recently been demonstrated that these two systems are primarily responsible for mitochondrial oxidation of cytosolic NADH in S. cerevisiae. Consistent with the hypothesis, the quadruple mutant grew on glucose as the sole carbon source. The growth on glucose, which was accompanied by glycerol production, was inhibited at high-glucose concentrations. This inhibition was attributed to glucose repression of respiratory enzymes as, in the quadruple mutant, respiratory pyruvate dissimilation is essential for ATP synthesis and growth. Serial transfer of the quadruple mutant on high-glucose media yielded a spontaneous mutant with much higher specific growth rates in high-glucose media (up to 0.10 h(-1) at 100 g of glucose. liter(-1)). In aerated batch cultures grown on 400 g of glucose. liter(-1), this engineered S. cerevisiae strain produced over 200 g of glycerol. liter(-1), corresponding to a molar yield of glycerol on glucose close to unity.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Overkamp</LastName><ForeName>Karin M</ForeName><Initials>KM</Initials><AffiliationInfo><Affiliation>Kluyver Laboratory of Biotechnology, Delft University of Technology, NL-2628 BC Delft, Amsterdam.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Bakker</LastName><ForeName>Barbara M</ForeName><Initials>BM</Initials></Author><Author ValidYN="Y"><LastName>Kötter</LastName><ForeName>Peter</ForeName><Initials>P</Initials></Author><Author ValidYN="Y"><LastName>Luttik</LastName><ForeName>Marijke A H</ForeName><Initials>MA</Initials></Author><Author ValidYN="Y"><LastName>Van Dijken</LastName><ForeName>Johannes P</ForeName><Initials>JP</Initials></Author><Author ValidYN="Y"><LastName>Pronk</LastName><ForeName>Jack T</ForeName><Initials>JT</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 Environ Microbiol</MedlineTA><NlmUniqueID>7605801</NlmUniqueID><ISSNLinking>0099-2240</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005994">Glycerophosphates</NameOfSubstance></Chemical><Chemical><RegistryNumber>0U46U6E8UK</RegistryNumber><NameOfSubstance UI="D009243">NAD</NameOfSubstance></Chemical><Chemical><RegistryNumber>9NTI6P3O4X</RegistryNumber><NameOfSubstance UI="C029620">alpha-glycerophosphoric acid</NameOfSubstance></Chemical><Chemical><RegistryNumber>IY9XDZ35W2</RegistryNumber><NameOfSubstance UI="D005947">Glucose</NameOfSubstance></Chemical><Chemical><RegistryNumber>PDC6A3C0OX</RegistryNumber><NameOfSubstance UI="D005990">Glycerol</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D018929" MajorTopicYN="N">Cell Culture Techniques</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003600" MajorTopicYN="N">Cytosol</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005818" MajorTopicYN="N">Genetic Engineering</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005947" MajorTopicYN="N">Glucose</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005990" MajorTopicYN="N">Glycerol</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005994" MajorTopicYN="N">Glycerophosphates</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008928" MajorTopicYN="N">Mitochondria</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009243" MajorTopicYN="N">NAD</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010084" MajorTopicYN="N">Oxidation-Reduction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010641" MajorTopicYN="N">Phenotype</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012441" MajorTopicYN="N">Saccharomyces cerevisiae</DescriptorName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName><QualifierName UI="Q000378" 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Bakker</name><name sortKey="Kotter, Peter" sort="Kotter, Peter" uniqKey="Kotter P" first="Peter" last="Kötter">Peter Kötter</name><name sortKey="Luttik, Marijke A H" sort="Luttik, Marijke A H" uniqKey="Luttik M" first="Marijke A H" last="Luttik">Marijke A H. Luttik</name><name sortKey="Pronk, Jack T" sort="Pronk, Jack T" uniqKey="Pronk J" first="Jack T" last="Pronk">Jack T. Pronk</name><name sortKey="Van Dijken, Johannes P" sort="Van Dijken, Johannes P" uniqKey="Van Dijken J" first="Johannes P" last="Van Dijken">Johannes P. Van Dijken</name></noCountry><country name="Pays-Bas"><region name="Hollande-Septentrionale"><name sortKey="Overkamp, Karin M" sort="Overkamp, Karin M" uniqKey="Overkamp K" first="Karin M" last="Overkamp">Karin M. Overkamp</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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|clé= pubmed:12039737
|texte= Metabolic engineering of glycerol production in Saccharomyces cerevisiae.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:12039737" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a DetoxFungiV1
| This area was generated with Dilib version V0.6.38. |  |