Abstract
BackgroundThe biomanufacturing of d-glucaric acid has attracted increasing interest because it is one of the top value-added chemicals produced from biomass. Saccharomyces cerevisiae is regarded as an excellent host for d-glucaric acid production.ResultsThe opi1 gene was knocked out because of its negative regulation on myo-inositol synthesis, which is the limiting step of d-glucaric acid production by S. cerevisiae. We then constructed the biosynthesis pathway of d-glucaric acid in S. cerevisiae INVSc1 opi1Δ and obtained two engineered strains, LGA-1 and LGA-C, producing record-breaking titers of d-glucaric acid: 9.53 ± 0.46 g/L and 11.21 ± 0.63 g/L d-glucaric acid from 30 g/L glucose and 10.8 g/L myo-inositol in fed-batch fermentation mode, respectively. However, LGA-1 was preferable because of its genetic stability and its superior performance in practical applications. There have been no reports on d-glucaric acid production from lignocellulose. Therefore, the biorefinery processes, including separated hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP) were investigated and compared. CBP using an artificial microbial consortium composed of Trichoderma reesei (T. reesei) Rut-C30 and S. cerevisiae LGA-1 was found to have relatively high d-glucaric acid titers and yields after 7 d of fermentation, 0.54 ± 0.12 g/L d-glucaric acid from 15 g/L Avicel and 0.45 ± 0.06 g/L d-glucaric acid from 15 g/L steam-exploded corn stover (SECS), respectively. In an attempt to design the microbial consortium for more efficient CBP, the team consisting of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be the best, with excellent work distribution and collaboration.ConclusionsTwo engineered S. cerevisiae strains, LGA-1 and LGA-C, with high titers of d-glucaric acid were obtained. This indicated that S. cerevisiae INVSc1 is an excellent host for d-glucaric acid production. Lignocellulose is a preferable substrate over myo-inositol. SHF, SSF, and CBP were studied, and CBP using an artificial microbial consortium of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be promising because of its relatively high titer and yield. T. reesei Rut-C30 and S. cerevisiae LGA-1were proven to be the best teammates for CBP. Further work should be done to improve the efficiency of this microbial consortium for d-glucaric acid production from lignocellulose.
Highlights
The biomanufacturing of d-glucaric acid has attracted increasing interest because it is one of the top value-added chemicals produced from biomass
Consolidated bioprocessing (CBP) using an artificial microbial consortium composed of Trichoderma reesei Rut-C30 and the engineered S. cerevisiae strain was established for direct production of d-glucaric acid from lignocelluloses
It was found that the deletion of opi1 promoted d-glucaric acid production, which was in line with a previous report [3]
Summary
The biomanufacturing of d-glucaric acid has attracted increasing interest because it is one of the top value-added chemicals produced from biomass. Li et al Biotechnol Biofuels (2021) 14:110 d-glucaric acid is produced via nitric acid oxidation of d-glucose. This is a nonselective and expensive process associated with a large exotherm, low yields and toxic byproducts [4,5,6]. A biosynthesis route from d-glucose to d-glucaric acid involving three heterologous genes was constructed in recombinant Escherichia coli by Moon et al [2]. A synthetic scaffold was used to increase MIOX stability and the efficiency of the biosynthesis route, leading to a titer of 2.5 g/L d-glucaric acid from 10 g/L d-glucose [7]. E. coli is thought to be unsuitable for d-glucaric acid production at high titers because d-glucaric acid concentrations above 5 g/L appear to inhibit E. coli through a pH-mediated effect [2, 3, 5]
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