1887

Abstract

Bio-production of fuels and chemicals from lignocellulosic C5 sugars usually requires the use of the pentose phosphate pathway (PPP) to produce pyruvate. Unfortunately, the oxidation of pyruvate to acetyl-coenzyme A results in the loss of 33 % of the carbon as CO, to the detriment of sustainability and process economics. To improve atom efficiency, we engineered to utilize -xylose constitutively using the Weimberg pathway, to allow direct production of 2-oxoglutarate without CO loss. After confirming enzyme expression , the pathway expression was optimized using a combinatorial approach, by screening a range of constitutive promoters whilst systematically varying the gene order. A PPP-deficient (), 2-oxoglutarate auxotroph () was used as the host strain, so that growth on xylose depended on the expression of the Weimberg pathway, and variants expressing could be selected on minimal agar plates. The strains were isolated and high-throughput measurement of the growth rates on -xylose was used to identify the fastest growing variant. This strain contained the pL promoter, with at the first position in the synthetic operon, and grew at 42 % of the rate on -xylose compared to wild-type using the PPP. Remarkably, the biomass yield was improved by 53.5 % compared with the wild-type upon restoration of activity. Therefore, the strain grows efficiently and constitutively on -xylose, and offers great potential for use as a new host strain to engineer carbon-efficient production of fuels and chemicals the Weimberg pathway.

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2018-03-01
2024-03-28
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