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Synthetic metabolic engineering-a novel, simple technology for designing a chimeric metabolic pathway

By Ye Xiaoting, Honda Kohsuke, Sakai Takaaki, Okano Kenji, Omasa Takeshi, Hirota Ryuichi, Kuroda Akio and Ohtake Hisao

Abstract

<p>Abstract</p> <p>Background</p> <p>The integration of biotechnology into chemical manufacturing has been recognized as a key technology to build a sustainable society. However, the practical applications of biocatalytic chemical conversions are often restricted due to their complexities involving the unpredictability of product yield and the troublesome controls in fermentation processes. One of the possible strategies to overcome these limitations is to eliminate the use of living microorganisms and to use only enzymes involved in the metabolic pathway. Use of recombinant mesophiles producing thermophilic enzymes at high temperature results in denaturation of indigenous proteins and elimination of undesired side reactions; consequently, highly selective and stable biocatalytic modules can be readily prepared. By rationally combining those modules together, artificial synthetic pathways specialized for chemical manufacturing could be designed and constructed.</p> <p>Results</p> <p>A chimeric Embden-Meyerhof (EM) pathway with balanced consumption and regeneration of ATP and ADP was constructed by using nine recombinant <it>E. coli</it> strains overproducing either one of the seven glycolytic enzymes of <it>Thermus thermophilus</it>, the cofactor-independent phosphoglycerate mutase of <it>Pyrococcus horikoshii</it>, or the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase of <it>Thermococcus kodakarensis</it>. By coupling this pathway with the <it>Thermus</it> malate/lactate dehydrogenase, a stoichiometric amount of lactate was produced from glucose with an overall ATP turnover number of 31.</p> <p>Conclusions</p> <p>In this study, a novel and simple technology for flexible design of a bespoke metabolic pathway was developed. The concept has been testified via a non-ATP-forming chimeric EM pathway. We designated this technology as “synthetic metabolic engineering”. Our technology is, in principle, applicable to all thermophilic enzymes as long as they can be functionally expressed in the host, and thus would be potentially applicable to the biocatalytic manufacture of any chemicals or materials on demand.</p

Topics: Microbiology, QR1-502
Publisher: BMC
Year: 2012
DOI identifier: 10.1186/1475-2859-11-120
OAI identifier: oai:doaj.org/article:0793d7dae4694f73a56a3125aba694dc
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