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dc.contributor.authorMacEachran, Daniel P.
dc.contributor.authorSinskey, Anthony J.
dc.date.accessioned2013-11-15T17:22:26Z
dc.date.available2013-11-15T17:22:26Z
dc.date.issued2013-11
dc.date.submitted2013-09
dc.identifier.issn1475-2859
dc.identifier.urihttp://hdl.handle.net/1721.1/82129
dc.description.abstractBackground: The Gram-positive actinomycete Rhodococcus opacus is widely studied for its innate ability to store large amounts of carbon in the form of triacylglycerol (TAG). Several groups have demonstrated that R. opacus PD630 is capable of storing anywhere from 50 to 76% of its cell dry weight as TAG. While numerous studies have focused on phenomenological aspects of this process, few have sought to identify the underlying molecular and biochemical mechanisms responsible for the biosynthesis and storage of this molecule. Results: Herein we further our previous efforts to illuminate the black box that is lipid metabolism in actinomycetes using a genetic approach. Utilizing a simple, colorimetric genetic screen, we have identified a gene, referred to herein as tadD (triacylglycerol accumulation deficient), which is critical for TAG biosynthesis in R. opacus PD630. Furthermore, we demonstrate that the purified protein product of this gene is capable of oxidizing glyceraldehyde-3-phosphate, while simultaneously reducing NAD(P)[superscript +] to NAD(P)H. Supporting this biochemical data, we observed that the ratio of NAD(P)H to NAD(P)[superscript +] is elevated in wildtype cultures grown under lipid production conditions as compared to cultures grown under vegetative growth conditions, while the mutant strain demonstrated no change irrespective of growth conditions. Finally, we demonstrate that over-expressing a putative phosphorylative glyceraldehyde-3-phosphate dehydrogenase leads to decreased TAG production during growth on TAG accumulation conditions. Conclusion: Taken together, the data support the identification of a key metabolic branch point separating vegetative growth and lipid accumulation lifestyles in Rhodococcus.en_US
dc.publisherBioMed Central Ltden_US
dc.relation.isversionofhttp://dx.doi.org/10.1186/1475-2859-12-104en_US
dc.rightsCreative Commons Attributionen_US
dc.rights.urihttp://creativecommons.org/licenses/by/2.0en_US
dc.sourceBioMed Central Ltden_US
dc.titleThe Rhodococcus opacus TadD protein mediates triacylglycerol metabolism by regulating intracellular NAD(P)H poolsen_US
dc.typeArticleen_US
dc.identifier.citationMacEachran, Daniel P, and Anthony J Sinskey. “The Rhodococcus Opacus TadD Protein Mediates Triacylglycerol Metabolism by Regulating Intracellular NAD(P)H Pools.” Microbial Cell Factories 12.1 (2013): 104.en_US
dc.contributor.departmentWhitaker College of Health Sciences and Technologyen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biologyen_US
dc.contributor.departmentMassachusetts Institute of Technology. Engineering Systems Divisionen_US
dc.contributor.mitauthorMacEachran, Daniel P.en_US
dc.contributor.mitauthorSinskey, Anthony J.en_US
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.updated2013-11-14T16:11:37Z
dc.language.rfc3066en
dc.rights.holderDaniel P MacEachran et al.; licensee BioMed Central Ltd.
dspace.orderedauthorsMacEachran, Daniel P; Sinskey, Anthony Jen_US
dc.identifier.orcidhttps://orcid.org/0000-0002-1015-1270
mit.licensePUBLISHER_CCen_US


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