Industrial Production of 2,3-Butanediol from the Engineered Corynebacterium glutamicum

Abstract

The platform chemical 2,3-butanediol (2,3-BDO) is a valuable product that can be converted into several petroleum-based chemicals via simple chemical reactions. Here, we produced 2,3-BDO with the non-pathogenic and rapidly growing Corynebacterium glutamicum. To enhance the 2,3-BDO production capacity of C. glutamicum, we introduced budA encoding acetolactate decarboxylase from Klebsiella pneumoniae, a powerful 2,3-BDO producer. Additionally, budB (encoding α-acetolactate synthase) and budC (encoding acetoin reductase) were introduced from K. pneumoniae to reinforce the carbon flux in the 2,3-BDO production. Because budC had a negative effect on 2,3-BDO production in C. glutamicum, the budB and budA introduced strain, SGSC102, was selected for 2,3-BDO production, and batch culture was performed at 30 °C, 250 rpm and pH 6.86 with pure glucose, molasses, and cassava powder as carbon substrates. After batch culture, significant amount of 2,3-BDO (18.9 and 12.0 g/L, respectively) was produced from 80 g/L of pure glucose and cassava powder.

Recent advances in metabolic engineering of Corynebacterium glutamicum as a potential platform microorganism for biorefinery

“…However, for l ‐lysine production, the yield from glucose was still significantly better than that from fructose (0.15 > 0.05 mol/mol) . The production of l ‐lysine, d ‐mannitol and 2,3‐butanediol from fructose using recombinant C. glutamicum have been demonstrated (Table ) …”

“…It can be also used as feedstock for biochemical production owing to its advantageous properties, including (i) high production, i.e., approximately 259 000 t in 2012, and (ii) convenient production of the important intermediate pyruvate via a one‐step enzymatic reaction mediated by lactate dehydrogenase . For the production of pyruvate‐derived platform chemicals, such as ethanol, isobutanol, and 2,3‐butanediol, utilization of lactic acid as feedstock is an effective strategy . C. glutamicum naturally utilizes lactate and harbors three genes involved in lactate metabolism, NCgl2816 (encoding lactate permease), lldD ( NCgl2817 ; encoding l ‐lactate dehydrogenases), and dld ( NCgl0865 ; encoding d ‐lactate dehydrogenase) (Fig.…”

“…Heterologous expression of the DEH degradation pathway in C. glutamicum has not been evaluated. The introduction of the proposed pathway into recombinant C. glutamicum should enable the increased production of pyruvate‐derived chemicals, such as ethanol, isobutanol, 2‐ketoisovalerate, and 2,3‐butanediol …”

“…Although C. glutamicum exhibits weak carbon catabolite repression toward mixed carbon sources, it cannot directly utilize complex polysaccharide polymers, such as xylan, cellulose, and starch, from lignocellulosic and starch biomass. To increase the economic feasibility of the biorefinery process, C. glutamicum has been engineered for the consolidated bioprocessing (CBP) of microalgal biomass and hemicellulose by the heterologous expression of amylolytic and cellulolytic enzymes . These recombinant C. glutamicum strains simultaneously hydrolyze biomass and produce biochemicals such as succinate, l ‐lysine, and xylonic acid using released glucose, maltose, cellobiose, d ‐xylose, and l ‐arabinose from biomass (Fig.…”

Evaluation of Brown Midrib Sorghum Mutants as a Potential Biomass Feedstock for 2,3-Butanediol Biosynthesis

“…A number of bacteria, including Klebsiella, Enterobacter, and Bacilli genera, produce 2,3butanediol from different sugar sources [4][5][6][7]. Most of the robust 2,3-butanediol-producing organisms reported so far, such as Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter aerogenes, and Serratia marcescens, belong to biosafety level 2 (pathogenic) [20,21]. The non-pathogenic (biosafety level 1) 2,3-butanediol producers such as Paenibacillus polymyxa, Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloliquefaciens are very low efficient compared to biosafety level 2 organism [4,20].…”