Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol

Abstract

Chirally pure (R)-1,3-butanediol ((R)-1,3-BDO) is a valuable intermediate for the production of fragrances, pheromones, insecticides and antibiotics. Biotechnological production results in superior enantiomeric excess over chemical production and is therefore the preferred production route. In this study (R)-1,3-BDO was produced in the industrially important whole cell biocatalyst Clostridium saccharoperbutylacetonicum through expression of the enantio-specific phaB gene from Cupriavidus necator. The heterologous pathway was optimised in three ways: at the transcriptional level choosing strongly expressed promoters and comparing plasmid borne with chromosomal gene expression, at the translational level by optimising the codon usage of the gene to fit the inherent codon adaptation index of C. saccharoperbutylacetonicum, and at the enzyme level by introducing point mutations which led to increased enzymatic activity. The resulting whole cell catalyst produced up to 20 mM (1.8 g/l) (R)-1,3-BDO in non-optimised batch fermentation which is a promising starting position for economical production of this chiral chemical.

Production of propionate using metabolically engineered strains of Clostridium saccharoperbutylacetonicum

“…In later stages of growth, FLU of all strains decreased as populations became non-fluorescent. Although P tcdB activity was higher when tcdR expression was under control of P bld , the lactose-inducible P bgaL promoter was chosen to establish propionate production in C. saccharoperbutylacetonicum to avoid possible pathway overloading due to the high activity observed for P bld (Grosse-Honebrink et al 2021 ). Furthermore, the use of P bgaL allows a controlled induction of gene expression and has recently successfully been used to establish a two-plasmid system carrying P tcdB and tcdR in C. saccharoperbutylacetonicum (Flaiz et al 2022 ).…”

“…Since it is a known hyper-butanol producer, it has mostly been employed for butanol production (Jiménez-Bonilla et al 2021 ). However, C. saccharoperbutylacetonicum has also successfully been used for the production of hydrogen (Singh et al 2019 ), isopropanol (Wang et al 2019 ), 1,3-butanediol (Grosse-Honebrink et al 2021 ), as well as caproate and hexanol (Wirth and Dürre 2021 ), thus highlighting its potential as a host for production of recombinant compounds. Here, we report the approach to convert C. saccharoperbutylacetonicum into a propionate producer by the implementation of the acrylate pathway from An.…”

Modulation of sol mRNA expression by the long non-coding RNA Assolrna in Clostridium saccharoperbutylacetonicum affects solvent formation

“…In addition to that, C . saccharoperbutylacetonicum can be used as production strain for other recombinant products like hexanol or 1,3-butanediol ( Grosse-Honebrink et al, 2021 ; Wirth and Dürre, 2021 ).…”

“…Acid production was recently optimized in C. saccharoperbutylacetonicum (Baur et al, 2022), since it can be used for large-scale production of high-value compounds such as butyl esters from sustainable resources (Feng et al, 2021). In addition to that, C. saccharoperbutylacetonicum can be used as production strain for other recombinant products like hexanol or 1,3-butanediol (Grosse-Honebrink et al, 2021;Wirth and Dürre, 2021).…”