Fermentative production of 1-propanol from d-glucose, l-rhamnose and glycerol using recombinant Escherichia coli

Abstract

Fermentative production of 1-propanol, which is one of the promising precursors of polypropylene production, from d-glucose, l-rhamnose and glycerol using metabolically engineered Escherichia coli was examined. To confer the ability to produce 1-propanol from 1,2-propanediol (1,2-PD) in recombinant E. coli, a part of the pdu regulon including the diol dehydratase and the propanol dehydrogenase genes together with the adenosylcobalamin (AdoCbl) regeneration enzyme genes of Klebsiella pneumoniae was cloned, and an expression vector for these genes (pRSF_pduCDEGHOQS) was constructed. Recombinant E. coli harboring pRSF_pduCDEGHOQS with 1,2-PD synthetic pathway (pKK_mde) genes, which was constructed in our previous report (Urano et al., Appl. Microbiol. Biotechnol., 99, 2001-2008, 2015), produced 16.1 mM of 1-propanol from d-glucose with a molar yield of 0.36 mol/mol after 72 h cultivation. 29.9 mM of 1-propanol was formed from l-rhamnose with a molar yield of 0.81 mol/mol using E. coli carrying only pRSF_pduCDEGHOQS. In addition, 1-propanol production from glycerol was achieved by addition of the ATP-dependent dihydroxyacetone kinase gene to E. coli harboring pKK_mde and pRSF_pduCDEGOQS. In all cases, 1-propanol production was achieved by adding only a small amount of AdoCbl.

Synthetic Biology and Metabolic Engineering Employing Escherichia coli for C2–C6 Bioalcohol Production

“…Several species of Clostridium have been evaluated for butanol and isopropanol production, but cannot be used for industrial application mainly due to low fermentation yield and titer ( Survase et al, 2011 ; Xue and Cheng, 2019 ). Alternative organisms such as E. coli have also been engineered toward the goal of industrial production of C3–C4 alcohols ( Jojima et al, 2008 ; Inokuma et al, 2010 ; Lan and Liao, 2013 ; Matsubara et al, 2016 ). E. coli metabolic pathways have shared intermediate metabolites, which reduce central metabolites such as acetyl-CoA and pyruvate into more electron-rich compounds and higher carbon acyl-CoA and 2-keto acids ( Saini et al, 2016 ; Heo et al, 2017 ; Ohtake et al, 2017 ; Soma et al, 2017 ; Nitta et al, 2019 ).…”

“…However, higher alcohols, except n-butanol, are not commonly produced at high yields in microbes. With the development of molecular biology techniques and metabolic engineering strategies, model systems, such as Escherichia coli (Jojima et al, 2008;Inokuma et al, 2010;Lan and Liao, 2013;Matsubara et al, 2016) and Saccharomyces cerevisiae (Park et al, 2014;Shi et al, 2016), have been modified to synthesize bioalcohols.…”

“…Escherichia coli is a well-studied model microorganism which has several advantageous traits for bioalcohol production including fast growth in inexpensive mineral media, the ability to utilize a wide range of substrates from biomass, and detailed genetic information and diverse genetic tools for gene manipulation. However, there are still challenges using engineered E. coli for industrial applications such as the need to improve tolerance to bioalcohols, efficient utilization of low-cost substrates, and productivity toward advanced alcohols (Jojima et al, 2008;Inokuma et al, 2010;Lan and Liao, 2013;Matsubara et al, 2016). Recently, the rapid expansion of genome engineering strategies, synthetic biology techniques, and high-throughput tools have enabled their application to study advanced bioalcohols production and to further investigate the mechanisms of alcohol resistance.…”

Transfer and analysis of Salmonella pdu genes in a range of Gram‐negative bacteria demonstrate exogenous microcompartment expression across a variety of species

“…In addition, such experiments allow an evolutionary analysis of pdu genes in which we can ask: Are the pdu genes from a given species expressed and functional in any other genera/species as opposed to having evolved to be restricted in these functions to the species of origin? Previous reports describing cloned Pdu MCP genes are present in the literature (Parsons et al ., ; Sargent et al ., ; Matsubara et al ., ). Sargent et al.…”

“…al. cloned a subset of the Klebsiella pneumoniae pdu genes and coupled them with a Shimwellia blatte 1,2 PD synthetic pathway in E. coli to achieve 1‐propanol production in this background (Matsubara et al ., ). In this study, we describe the first cloning of the entire, contiguous S .…”