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dc.contributor.authorZhou, Kang
dc.contributor.authorZou, Ruiyang
dc.contributor.authorStephanopoulos, Gregory
dc.contributor.authorToo, Heng-Phon
dc.date.accessioned2013-01-23T21:57:41Z
dc.date.available2013-01-23T21:57:41Z
dc.date.issued2012-11
dc.identifier.issn1475-2859
dc.identifier.urihttp://hdl.handle.net/1721.1/76588
dc.description.abstractBackground: Recombinant proteins are routinely overexpressed in metabolic engineering. It is well known that some over-expressed heterologous recombinant enzymes are insoluble with little or no enzymatic activity. This study examined the solubility of over-expressed homologous enzymes of the deoxyxylulose phosphate pathway (DXP) and the impact of inclusion body formation on metabolic engineering of microbes. Results: Four enzymes of this pathway (DXS, ISPG, ISPH and ISPA), but not all, were highly insoluble, regardless of the expression systems used. Insoluble dxs (the committed enzyme of DXP pathway) was found to be inactive. Expressions of fusion tags did not significantly improve the solubility of dxs. However, hypertonic media containing sorbitol, an osmolyte, successfully doubled the solubility of dxs, with the concomitant improvement in microbial production of the metabolite, DXP. Similarly, sorbitol significantly improved the production of soluble and functional ERG12, the committed enzyme in the mevalonate pathway. Conclusion: This study demonstrated the unanticipated findings that some over-expressed homologous enzymes of the DXP pathway were highly insoluble, forming inclusion bodies, which affected metabolite formation. Sorbitol was found to increase both the solubility and function of some of these over-expressed enzymes, a strategy to increase the production of secondary metabolites.en_US
dc.description.sponsorshipSingapore–MIT Alliance for Research and Technology (Flagship Research Project)en_US
dc.publisherBioMed Central Ltd.en_US
dc.relation.isversionofhttp://dx.doi.org/10.1186/1475-2859-11-148en_US
dc.rightsCreative Commons Attributionen_US
dc.rights.urihttp://creativecommons.org/licenses/by/2.0en_US
dc.sourceBioMed Central Ltden_US
dc.titleEnhancing solubility of deoxyxylulose phosphate pathway enzymes for microbial isoprenoid productionen_US
dc.typeArticleen_US
dc.identifier.citationZhou, Kang et al. “Enhancing Solubility of Deoxyxylulose Phosphate Pathway Enzymes for Microbial Isoprenoid Production.” Microbial Cell Factories 11.1 (2012): 148. Web.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemical Engineeringen_US
dc.contributor.departmentSingapore-MIT Alliance in Research and Technology (SMART)en_US
dc.contributor.mitauthorZhou, Kang
dc.contributor.mitauthorZou, Ruiyang
dc.contributor.mitauthorStephanopoulos, Gregory
dc.contributor.mitauthorToo, Heng-Phon
dc.relation.journalMicrobial Cell Factoriesen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2013-01-15T16:05:27Z
dc.language.rfc3066en
dc.rights.holderKang Zhou et al.; licensee BioMed Central Ltd.
dspace.orderedauthorsZhou, Kang; Zou, Ruiyang; Stephanopoulos, Gregory; Too, Heng-Phonen
dc.identifier.orcidhttps://orcid.org/0000-0001-6909-4568
mit.licensePUBLISHER_CCen_US
mit.metadata.statusComplete


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