2,3-Butanediol production from cellobiose by engineered Saccharomyces cerevisiae

Hong Nan, Seung Oh Seo, Eun Joong Oh, Jin Ho Seo, Jamie H.D. Cate, Yong Su Jin

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Production of renewable fuels and chemicals from cellulosic biomass is a critical step towards energy sustainability and reduced greenhouse gas emissions. Microbial cells have been engineered for producing chemicals from cellulosic sugars. Among these chemicals, 2,3-butanediol (2,3-BDO) is a compound of interest due to its diverse applications. While microbial production of 2,3-BDO with high yields and productivities has been reported, there are concerns associated with utilization of potential pathogenic bacteria and inefficient utilization of cellulosic sugars. To address these problems, we engineered 2,3-BDO production in Saccharomyces cerevisiae, especially from cellobiose, a prevalent sugar in cellulosic hydrolysates. Specifically, we overexpressed alsS and alsD from Bacillus subtilis to convert pyruvate into 2,3-BDO via α-acetolactate and acetoin in an engineered cellobiose fermenting S. cerevisiae. Under oxygen-limited conditions, the resulting strain was able to produce 2,3-BDO. Still, major carbon flux went to ethanol, resulting in substantial amounts of ethanol produced as a byproduct. To enhance pyruvate flux to 2,3-BDO through elimination of the pyruvate decarboxylation reaction, we employed a deletion mutant of both PDC1 and PDC5 for producing 2,3-BDO. When a cellobiose utilization pathway, consisting of a cellobiose transporter and intracellular β-glucosidase, and the 2,3-BDO producing pathway were introduced in a pyruvate decarboxylase deletion mutant, the resulting strain produced 2,3-BDO without ethanol production from cellobiose under oxygen-limited conditions. A titer of 5.29 g/l 2,3-BDO with a productivity of 0.22 g/l h and yield of 0.29 g 2,3-BDO/g cellobiose was attained. These results suggest the possibility of producing 2,3-BDO safely and sustainably from cellulosic hydrolysates.

Original languageEnglish (US)
Pages (from-to)5757-5764
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume98
Issue number12
DOIs
StatePublished - Jun 2014

Keywords

  • 2,3-Butanediol
  • Cellobiose
  • Pyruvate decarboxylase deletion
  • S. cerevisiae

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