Abstract

1,2-propanediol (1,2-PDO) is a versatile chemical used in multiple manufacturing processes. To date, some engineered and non-engineered microbes, such as Escherichia coli, Lactobacillus buchneri, and Clostridium thermosaccharolyticum, have been used to produce 1,2-PDO. In this study, we demonstrated the production of R- and S-1,2-PDO using engineered Lactococcus lactis. The L- and D-lactic acid-producing L. lactis strains NZ9000 and AH1 were transformed with the plasmid pNZ8048-ppy harboring pct, pduP, and yahK genes for 1,2-PDO biosynthesis, resulting in L. lactis LL1 and LL2, respectively. These engineered L. lactis produced S- and R-1,2-PDO at concentrations of 0.69 and 0.50 g/L with 94.4 and 78.0% ee optical purities, respectively, from 1% glucose after 72 h of cultivation. Both 1% mannitol and 1% gluconate were added instead of glucose to the culture of L. lactis LL1 to supply NADH and NADPH to the 1,2-PDO production pathway, resulting in 75% enhancement of S-1,2-PDO production. Production of S-1,2-PDO from 5% mannitol and 5% gluconate was demonstrated using L. lactis LL1 with a pH–stat approach. This resulted in S-1,2-PDO production at a concentration of 1.88 g/L after 96 h of cultivation. To our knowledge, this is the first report on the production of R- and S-1,2-PDO using engineered lactic acid bacteria.

Highlights

  • Sustainable manufacturing from biomass is in great demand to minimize negative environmental impacts (Abbasi and Abbasi 2010; Usmani et al 2021)

  • Since the biosynthetic pathway for the production of 1,2-PDO has been reported, bioproduction of this compound has been attempted by introducing the biosynthetic pathway in various microorganisms including E. coli (Cameron and Cooney 1986, Niu and Guo 2015, Niu et al 2019)

  • We demonstrated production of R- and S-1,2-PDO using engineered L. lactis constructed from L- and D-lactic acid-producing lactic acid bacteria (LAB), L. lactis NZ9000, and AH1, respectively, by expression of 1,2-PDO biosynthetic genes with glucose, mannitol, and/ or gluconate (Fig. 1)

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Summary

Introduction

Sustainable manufacturing from biomass is in great demand to minimize negative environmental impacts (Abbasi and Abbasi 2010; Usmani et al 2021). 1,2-PDO can be produced in engineered microbes via a pathway in which glucose is first converted to D- and L-lactate, followed by the synthesis of R- and S-1,2-PDO from D- and L-lactate, respectively, in which cofactors such as acetyl-CoA, NADH, and NADPH are required (Altras and Cameron 1999; Saxena et al 2010; Niu and Guo 2015; Niu et al 2019) This suggests that microbes producing the 1,2-PDO precursor lactate at a high titer are suitable as production hosts for 1,2-PDO production, and that R- and S-1,2-PDO can be separately produced in D- and L-lactic acid producers, respectively.

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