Biosynthesis of chiral 3-hydroxyvalerate from single propionate-unrelated carbon sources in metabolically engineered E. coli

Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Microbial Cell Factories (Impact Factor: 4.22). 11/2010; 9(1):96. DOI: 10.1186/1475-2859-9-96
Source: PubMed


The ability to synthesize chiral building block molecules with high optical purity is of considerable importance to the fine chemical and pharmaceutical industries. Production of one such compound, 3-hydroxyvalerate (3HV), has previously been studied with respect to the in vivo or in vitro enzymatic depolymerization of biologically-derived co-polymers of poly(3-hydroxybutyrate-co-3-hydroxyvalerate). However, production of this biopolymeric precursor typically necessitates the supplementation of a secondary carbon source (e.g., propionate) into the culture medium. In addition, previous approaches for producing 3HV have not focused on its enantiopure synthesis, and thus suffer from increased costs for product purification.
Here, we report the selective biosynthesis of each 3HV stereoisomer from a single, renewable carbon source using synthetic metabolic pathways in recombinant strains of Escherichia coli. The product chirality was controlled by utilizing two reductases of opposing stereoselectivity. Improvement of the biosynthetic pathway activity and host background was carried out to elevate both the 3HV titers and 3HV/3HB ratios. Overall, shake-flask titers as high as 0.31 g/L and 0.50 g/L of (S)-3HV and (R)-3HV, respectively, were achieved in glucose-fed cultures, whereas glycerol-fed cultures yielded up to 0.19 g/L and 0.96 g/L of (S)-3HV and (R)-3HV, respectively.
Our work represents the first report of direct microbial production of enantiomerically pure 3HV from a single carbon source. Continued engineering of host strains and pathway enzymes will ultimately lead to more economical production of chiral 3HV.

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Available from: Hsien-Chung Tseng, Mar 06, 2014
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    • "Besides the use of Ter, butyrate production by E. coli was substantially enhanced by genetically deleting competing native fermentation pathways (Lim et al., 2013) as was previously done for the fermentative production of ethanol (Jarboe et al., 2007) and other bio-based products (Lim et al., 2013; Shen et al., 2011; Zhu et al., 2011). In addition to butyrate production, reactions similar to those described above for the conversion of two molecules of acetyl-CoA to butyryl-CoA together with TesB have been used under aerobic conditions to produce functionalized short-chain carboxylic acids, including 3-hydroxyvalerate (3-hydroxy- pentanoate) (Tseng et al., 2010), dihydroxybutyrate (Martin et al., 2013), 3-hydroxy-4-methylvalerate (Martin et al., 2013) and a variety of alcohols (Tseng & Prather, 2012). Similar to reverse b-oxidation, fermentative pathways proceed by sequential condensation of acetyl-CoA and are more energy efficient than the interruption of phospholipid biosynthesis. "
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    • "(S)-3HB biomonomer, meanwhile, can be used to produce novel PHBs with new features and properties. Prather and coworkers subsequently expanded upon their efforts to produce, for the first time, enantiomerically pure stereoisomers of the 5-carbon β-hydroxyacid 3HV (Tseng et al., 2010). This was achieved using a single renewable substrate (glucose) by first engineering E. coli to over-produce precursor propionyl-CoA via manipulation of the native L-threonine biosynthesis pathway. "
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