caa a22 4500 606212426 CHVBK 20210128101042.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s10295-015-1589-6 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1589-6 Synthesis of medium-chain length (C6-C10) fuels and chemicals via β-oxidation reversal in Escherichia coli [Elektronische Daten] [Seohyoung Kim, James Clomburg, Ramon Gonzalez] The recently engineered reversal of the β-oxidation cycle has been proposed as a potential platform for the efficient synthesis of longer chain (C≥4) fuels and chemicals. Here, we demonstrate the utility of this platform for the synthesis of medium-chain length (C6-C10) products through the manipulation of key components of the pathway. Deletion of endogenous thioesterases provided a clean background in which the expression of various thiolase and termination components, along with required core enzymes, resulted in the ability to alter the chain length distribution and functionality of target products. This approach enabled the synthesis of medium-chain length carboxylic acids and primary alcohols from glycerol, a low-value feedstock. The use of BktB as the thiolase component with thioesterase TesA' as the termination enzyme enabled the synthesis of about 1.3g/L C6-C10 saturated carboxylic acids. Tailoring of product formation to primary alcohol synthesis was achieved with the use of various acyl-CoA reductases. The combination of AtoB and FadA as the thiolase components with the alcohol-forming acyl-CoA reductase Maqu2507 from M. aquaeolei resulted in the synthesis of nearly 0.3g/L C6-C10 alcohols. These results further demonstrate the versatile nature of a β-oxidation reversal, and highlight several key aspects and control points that can be further manipulated to fine-tune the synthesis of various fuels and chemicals. Society for Industrial Microbiology and Biotechnology, 2015 β-Oxidation reversal nationallicence Metabolic engineering nationallicence Synthetic biology nationallicence Fuels and chemicals nationallicence Medium-chain acids nationallicence Medium-chain alcohols nationallicence Kim Seohyoung Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, 77005, Houston, TX, USA aut Clomburg James Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, 77005, Houston, TX, USA aut Gonzalez Ramon Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, 77005, Houston, TX, USA aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/3(2015-03-01), 465-475 1367-5435 42:3<465 2015 42 10295 https://doi.org/10.1007/s10295-015-1589-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1589-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kim Seohyoung Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, 77005, Houston, TX, USA aut NATIONALLICENCE 700 1- Clomburg James Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, 77005, Houston, TX, USA aut NATIONALLICENCE 700 1- Gonzalez Ramon Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, 77005, Houston, TX, USA aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/3(2015-03-01), 465-475 1367-5435 42:3<465 2015 42 10295