Cell-free protein synthesis energized by slowly-metabolized maltodextrin

Wang, Yiran; Zhang, Y. H. Percival

Cell-free protein synthesis energized by slowly-metabolized maltodextrin

dc.contributor.author	Wang, Yiran	en
dc.contributor.author	Zhang, Y. H. Percival	en
dc.contributor.department	Biological Systems Engineering	en
dc.contributor.department	Institute for Critical Technology and Applied Science	en
dc.date.accessioned	2012-08-24T12:00:23Z	en
dc.date.available	2012-08-24T12:00:23Z	en
dc.date.issued	2009-06-28	en
dc.date.updated	2012-08-24T12:00:23Z	en
dc.description.abstract	Background 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.version	Published version	en
dc.format.mimetype	application/pdf	en
dc.identifier.citation	BMC Biotechnology. 2009 Jun 28;9(1):58	en
dc.identifier.doi	https://doi.org/10.1186/1472-6750-9-58	en
dc.identifier.uri	http://hdl.handle.net/10919/18895	en
dc.language.iso	en	en
dc.rights	Creative Commons Attribution 4.0 International	en
dc.rights.holder	Yiran Wang et al.; licensee BioMed Central Ltd.	en
dc.rights.uri	http://creativecommons.org/licenses/by/4.0/	en
dc.title	Cell-free protein synthesis energized by slowly-metabolized maltodextrin	en
dc.title.serial	BMC Biotechnology	en
dc.type	Article - Refereed	en
dc.type.dcmitype	Text	en