Engineering Escherichia coli for Direct Production of 1,2-Propanediol and 1,3-Propanediol from Starch
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Engineering Escherichia coli for Direct Production of 1,2‑Propanediol and 1,3‑Propanediol from Starch Rintaro Sato1,2 · Tomonari Tanaka1 · Hitomi Ohara1 · Yuji Aso1,2 Received: 28 April 2020 / Accepted: 26 August 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Diols are versatile chemicals used for multiple manufacturing products. In some previous studies, Escherichia coli has been engineered to produce 1,2-propanediol (1,2-PDO) and 1,3-propanediol (1,3-PDO) from glucose. However, there are no reports on the direct production of these diols from starch instead of glucose as a substrate. In this study, we directly produced 1,2-PDO and 1,3-PDO from starch using E. coli engineered for expressing a heterologous α-amylase, along with the expression of 1,2-PDO and 1,3-PDO synthetic genes. For this, the recombinant plasmids, pVUB3-SBA harboring amyA gene for α-amylase production, pSR5 harboring pct, pduP, and yahK genes for 1,2-PDO production, and pSR8 harboring gpd1-gpp2, dhaB123, gdrAB, and dhaT genes for 1,3-PDO production, were constructed. Subsequently, E. coli BW25113 (ΔpflA) and BW25113 strains were transformed with pVUB3-SBA, pSR5, and/or pSR8. Using these transformants, direct production of 1,2-PDO and 1,3-PDO from starch was demonstrated under microaerobic condition. As a result, the maximum production titers of 1,2-PDO and 1,3-PDO from 1% glucose as a sole carbon source were 13 mg/L and 150 mg/L, respectively. The maximum production titers from 1% starch were similar levels (30 mg/L 1,2-PDO and 120 mg/L 1,3-PDO). These data indicate that starch can be an alternative carbon source for the production of 1,2-PDO and 1,3-PDO in engineered E. coli. This technology could simplify the upstream process of diol bioproduction.
Introduction Biorefining is a technology that produces biobased fuels and chemicals from primarily starch-based crops such as corn [1, 2]. Since diols are versatile compounds harboring two hydroxyl groups, they are used as platform chemicals for many industrial applications. Biobased diols are promising targets produced from biomass through the biorefinery process [3]. Among biobased diols, 1,3-propanediol (1,3-PDO) has enabled synthesis of multiple synthetic polymers such as new biobased polyester, poly(trimethylene terephthalate) [4, 5]. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00284-020-02189-8) contains supplementary material, which is available to authorized users. * Yuji Aso [email protected] 1
Department of Biobased Materials Science, Kyoto Institute of Technology, Kyoto, Japan
JST‑Mirai Program, Japan Science and Technology Agency, Saitama, Japan
2
Both 1,2-PDO and 1,3-PDO are C3 diols that can be produced by engineered and non-engineered microbes, using glucose, glycerol, or xylose as the carbon source [6–8]. Clostridium spp. and Corynebacterium glutamicum produce 1,2-PDO from glucose or xylose [9–12]. Clostridium acetobutylicum and ` produce 1,3-PDO from glycerol [13–15]. Escherichia coli
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