Cell-free protein synthesis energized by slowly-metabolized maltodextrin

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dc.contributor.authorWang, Yiranen
dc.contributor.authorZhang, Y. H. Percivalen
dc.contributor.departmentBiological Systems Engineeringen
dc.contributor.departmentInstitute for Critical Technology and Applied Scienceen
dc.date.accessioned2012-08-24T12:00:23Zen
dc.date.available2012-08-24T12:00:23Zen
dc.date.issued2009-06-28en
dc.date.updated2012-08-24T12:00:23Zen
dc.description.abstractBackground Cell-free protein synthesis (CFPS) is a rapid and high throughput technology for obtaining proteins from their genes. The primary energy source ATP is regenerated from the secondary energy source through substrate phosphorylation in CFPS. Results Distinct from common secondary energy sources (e.g., phosphoenolpyruvate - PEP, glucose-6-phosphate), maltodextrin was used for energizing CFPS through substrate phosphorylation and the glycolytic pathway because (i) maltodextrin can be slowly catabolized by maltodextrin phosphorylase for continuous ATP regeneration, (ii) maltodextrin phosphorylation can recycle one phosphate per reaction for glucose-1-phosphate generation, and (iii) the maltodextrin chain-shortening reaction can produce one ATP per glucose equivalent more than glucose can. Three model proteins, esterase 2 from Alicyclobacillus acidocaldarius, green fluorescent protein, and xylose reductase from Neurospora crassa were synthesized for demonstration. Conclusion Slowly-metabolized maltodextrin as a low-cost secondary energy compound for CFPS produced higher levels of proteins than PEP, glucose, and glucose-6-phospahte. The enhancement of protein synthesis was largely attributed to better-controlled phosphate levels (recycling of inorganic phosphate) and a more homeostatic reaction environment.en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationBMC Biotechnology. 2009 Jun 28;9(1):58en
dc.identifier.doihttps://doi.org/10.1186/1472-6750-9-58en
dc.identifier.urihttp://hdl.handle.net/10919/18895en
dc.language.isoenen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.holderYiran Wang et al.; licensee BioMed Central Ltd.en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.titleCell-free protein synthesis energized by slowly-metabolized maltodextrinen
dc.title.serialBMC Biotechnologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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