RT Journal Article
SR Electronic
T1 Discovery and Evaluation of Biosynthetic Pathways for the Production of Five Methyl Ethyl Ketone Precursors
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 209569
DO 10.1101/209569
A1 Milenko Tokic
A1 Noushin Hadadi
A1 Meric Ataman
A1 Dario S. Neves
A1 Birgitta E. Ebert
A1 Lars M. Blank
A1 Ljubisa Miskovic
A1 Vassily Hatzimanikatis
YR 2017
UL http://biorxiv.org/content/early/2017/10/26/209569.abstract
AB The limited supply of fossil fuels and the establishment of new environmental policies shifted research in industry and academia towards sustainable production of the 2nd generation of biofuels, with Methyl Ethyl Ketone (MEK) being one promising fuel candidate. MEK is a commercially valuable petrochemical with an extensive application as a solvent. However, as of today, a sustainable and economically viable production of MEK has not yet been achieved despite several attempts of introducing biosynthetic pathways in industrial microorganisms. We used BNICE.ch to discover all novel pathways around MEK. Out of 1’325 identified compounds connecting to MEK with one reaction step, we selected 3-oxopentanoate, but-3-en-2-one, but-1-en-2-olate, butylamine, and 2-hydroxy-2-methyl-butanenitrile for further study. We reconstructed 3’679’610 novel biosynthetic pathways towards these 5 compounds. We then embedded these pathways into the genome-scale model of E. coli, retaining a set of 18’925 most biologically viable ones based on their thermodynamic feasibilities and yields. For each novel reaction in the viable pathways, we proposed the most similar KEGG reactions, with their gene and protein sequences, as candidates for either a direct experimental implementation or as basis for enzyme engineering. Through pathway similarity analysis we classified the pathways and identified the enzymes and precursors that were indispensable for the production of the target molecules. The developments from this study enhance the potential of BNICE.ch for discovery, systematic evaluation, and analysis of novel pathways in future synthetic biology and metabolic engineering studies.Graphical abstract