Production of the sesquiterpene (+)-valencene by metabolically engineered Corynebacterium glutamicum.
Identifieur interne : 000953 ( Main/Exploration ); précédent : 000952; suivant : 000954Production of the sesquiterpene (+)-valencene by metabolically engineered Corynebacterium glutamicum.
Auteurs : Jonas Frohwitter [Allemagne] ; Sabine A E. Heider [Allemagne] ; Petra Peters-Wendisch [Allemagne] ; Jules Beekwilder [Pays-Bas] ; Volker F. Wendisch [Allemagne]Source :
- Journal of biotechnology [ 1873-4863 ] ; 2014.
English descriptors
- KwdEn :
- Alkanes (metabolism), Amino Acid Sequence (genetics), Carotenoids (biosynthesis), Carotenoids (genetics), Citrus (chemistry), Corynebacterium glutamicum (enzymology), Corynebacterium glutamicum (genetics), Escherichia coli, Escherichia coli Proteins (genetics), Escherichia coli Proteins (metabolism), Geranyltranstransferase, Metabolic Engineering, Polyisoprenyl Phosphates (biosynthesis), Polyisoprenyl Phosphates (metabolism), Sequence Alignment, Sesquiterpenes (chemistry), Sesquiterpenes (metabolism).
- MESH :
- chemical , biosynthesis : Carotenoids, Polyisoprenyl Phosphates.
- chemical , chemistry : Sesquiterpenes.
- chemical , genetics : Carotenoids, Escherichia coli Proteins.
- chemical , metabolism : Alkanes, Escherichia coli Proteins, Polyisoprenyl Phosphates, Sesquiterpenes.
- chemistry : Citrus.
- enzymology : Corynebacterium glutamicum.
- genetics : Amino Acid Sequence, Corynebacterium glutamicum.
- Escherichia coli, Geranyltranstransferase, Metabolic Engineering, Sequence Alignment.
Abstract
The sesquiterpene (+)-valencene is an aroma compound of citrus fruits and is used to flavor foods and drinks. Biosynthesis of (+)-valencene starts from farnesyl pyrophosphate, an intermediate of carotenoid biosynthesis. Corynebacterium glutamicum, the workhorse of the million-ton scale amino acid industry, is naturally pigmented as it synthesizes the rare fifty carbon atoms (C50) containing carotenoid decaprenoxanthin. Since the carotenoid pathway of this Gram-positive bacterium has previously been engineered for efficient production of several C50 and C40 carotenoids, its potential to produce a sesquiterpene was assessed. Growth of C. glutamicum was negatively affected by (+)-valencene, but overlaying n-dodecane as organic phase for extraction of (+)-valencene was shown to be biocompatible. Heterologous expression of the (+)-valencene synthase gene from the sweet orange Citrus sinensis was not sufficient to enable (+)-valencene production, likely because provision of farnesyl pyrophosphate (FPP) by endogenous prenyltransferases was too low. However, upon deletion of two endogenous prenyltransferase genes and heterologous expression of either FPP synthase gene ispA from Escherichia coli or ERG20 from Saccharomyces cerevisiae (+)-valence production by C. sinensis valencene synthase was observed. Employing the valencene synthase from Nootka cypress improved (+)-valencene titers 10 fold to 2.41±0.26mgl(-1) (+)-valencene, which is equivalent to 0.25±0.03mgg(-1) cell dry weight (CDW). This is the first report on sesquiterpene overproduction by recombinant C. glutamicum.
DOI: 10.1016/j.jbiotec.2014.05.032
PubMed: 24910970
Affiliations:
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Heider</name><affiliation wicri:level="1"><nlm:affiliation>Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld 33615, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld 33615</wicri:regionArea><wicri:noRegion>33615</wicri:noRegion><wicri:noRegion>33615</wicri:noRegion><wicri:noRegion>Bielefeld 33615</wicri:noRegion></affiliation></author><author><name sortKey="Peters Wendisch, Petra" sort="Peters Wendisch, Petra" uniqKey="Peters Wendisch P" first="Petra" last="Peters-Wendisch">Petra Peters-Wendisch</name><affiliation wicri:level="1"><nlm:affiliation>Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld 33615, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld 33615</wicri:regionArea><wicri:noRegion>33615</wicri:noRegion><wicri:noRegion>33615</wicri:noRegion><wicri:noRegion>Bielefeld 33615</wicri:noRegion></affiliation></author><author><name sortKey="Beekwilder, Jules" sort="Beekwilder, Jules" uniqKey="Beekwilder J" first="Jules" last="Beekwilder">Jules Beekwilder</name><affiliation wicri:level="1"><nlm:affiliation>Plant Research International, Wageningen, The Netherlands.</nlm:affiliation><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Plant Research International, Wageningen</wicri:regionArea><wicri:noRegion>Wageningen</wicri:noRegion></affiliation></author><author><name sortKey="Wendisch, Volker F" sort="Wendisch, Volker F" uniqKey="Wendisch V" first="Volker F" last="Wendisch">Volker F. Wendisch</name><affiliation wicri:level="1"><nlm:affiliation>Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld 33615, Germany. Electronic address: volker.wendisch@uni-bielefeld.de.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Bielefeld 33615</wicri:regionArea><wicri:noRegion>33615</wicri:noRegion><wicri:noRegion>33615</wicri:noRegion><wicri:noRegion>Bielefeld 33615</wicri:noRegion></affiliation></author></analytic><series><title level="j">Journal of biotechnology</title><idno type="eISSN">1873-4863</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Alkanes (metabolism)</term><term>Amino Acid Sequence (genetics)</term><term>Carotenoids (biosynthesis)</term><term>Carotenoids (genetics)</term><term>Citrus (chemistry)</term><term>Corynebacterium glutamicum (enzymology)</term><term>Corynebacterium glutamicum (genetics)</term><term>Escherichia coli</term><term>Escherichia coli Proteins (genetics)</term><term>Escherichia coli Proteins (metabolism)</term><term>Geranyltranstransferase</term><term>Metabolic Engineering</term><term>Polyisoprenyl Phosphates (biosynthesis)</term><term>Polyisoprenyl Phosphates (metabolism)</term><term>Sequence Alignment</term><term>Sesquiterpenes (chemistry)</term><term>Sesquiterpenes (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Carotenoids</term><term>Polyisoprenyl Phosphates</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Sesquiterpenes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Carotenoids</term><term>Escherichia coli Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Alkanes</term><term>Escherichia coli Proteins</term><term>Polyisoprenyl Phosphates</term><term>Sesquiterpenes</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Citrus</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Corynebacterium glutamicum</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Amino Acid Sequence</term><term>Corynebacterium glutamicum</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Escherichia coli</term><term>Geranyltranstransferase</term><term>Metabolic Engineering</term><term>Sequence Alignment</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The sesquiterpene (+)-valencene is an aroma compound of citrus fruits and is used to flavor foods and drinks. Biosynthesis of (+)-valencene starts from farnesyl pyrophosphate, an intermediate of carotenoid biosynthesis. Corynebacterium glutamicum, the workhorse of the million-ton scale amino acid industry, is naturally pigmented as it synthesizes the rare fifty carbon atoms (C50) containing carotenoid decaprenoxanthin. Since the carotenoid pathway of this Gram-positive bacterium has previously been engineered for efficient production of several C50 and C40 carotenoids, its potential to produce a sesquiterpene was assessed. Growth of C. glutamicum was negatively affected by (+)-valencene, but overlaying n-dodecane as organic phase for extraction of (+)-valencene was shown to be biocompatible. Heterologous expression of the (+)-valencene synthase gene from the sweet orange Citrus sinensis was not sufficient to enable (+)-valencene production, likely because provision of farnesyl pyrophosphate (FPP) by endogenous prenyltransferases was too low. However, upon deletion of two endogenous prenyltransferase genes and heterologous expression of either FPP synthase gene ispA from Escherichia coli or ERG20 from Saccharomyces cerevisiae (+)-valence production by C. sinensis valencene synthase was observed. Employing the valencene synthase from Nootka cypress improved (+)-valencene titers 10 fold to 2.41±0.26mgl(-1) (+)-valencene, which is equivalent to 0.25±0.03mgg(-1) cell dry weight (CDW). This is the first report on sesquiterpene overproduction by recombinant C. glutamicum.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>Pays-Bas</li></country></list><tree><country name="Allemagne"><noRegion><name sortKey="Frohwitter, Jonas" sort="Frohwitter, Jonas" uniqKey="Frohwitter J" first="Jonas" last="Frohwitter">Jonas Frohwitter</name></noRegion><name sortKey="Heider, Sabine A E" sort="Heider, Sabine A E" uniqKey="Heider S" first="Sabine A E" last="Heider">Sabine A E. Heider</name><name sortKey="Peters Wendisch, Petra" sort="Peters Wendisch, Petra" uniqKey="Peters Wendisch P" first="Petra" last="Peters-Wendisch">Petra Peters-Wendisch</name><name sortKey="Wendisch, Volker F" sort="Wendisch, Volker F" uniqKey="Wendisch V" first="Volker F" last="Wendisch">Volker F. Wendisch</name></country><country name="Pays-Bas"><noRegion><name sortKey="Beekwilder, Jules" sort="Beekwilder, Jules" uniqKey="Beekwilder J" first="Jules" last="Beekwilder">Jules Beekwilder</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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