Recruiting Energy-Conserving Sucrose Utilization Pathways for Enhanced 2,3-Butanediol Production in Bacillus subtilis

Abstract

To improve the utilization of sucrose for 2,3-butanediol (2,3-BD) production, four combinations of heterologous energy-conserving sucrose utilization pathways were introduced into Bacillus subtilis Δtet strain (a derivative from B. subtilis 168) and B. subtilis FJ-1 strain (a new isolate in our lab). Results demonstrated that the combination of cscB (encoding sucrose permease) from Escherichia coli and gtfA (encoding sucrose phosphorylase) from Streptococcus mutans showed the most remarkable enhancement for the 2,3-BD production. With sucrose and sugar cane juice as substrate, respectively, the Δtet-CEG strain (B. subtilis Δtet strain containing the energy-conserving sucrose utilization pathway as referred above) showed 36.5% and 24.7% increase in 2,3-BD production than the control Δtet strain, and the FJ-1-CEG strain also produced 23.8% and 44.5% more 2,3-BD than the control FJ-1 strain. The metabolic engineering strategy demonstrated in this study can be extensively applied to other microorganisms for reinforced production of desirable biochemicals from sucrose. Meanwhile, the newly isolated FJ-1 strain is an interesting platform strain that can be further engineered for efficient 2,3-BD production.

Simultaneous production of poly-γ-glutamic acid and 2,3-butanediol by a newly isolated Bacillus subtilis CS13

“…In the utilization of sucrose, B. subtilis NX-2 showed a good advantage to improve the γ-PGA production [16]. Additionally, sucrose was a popular carbon source for production of 2,3-BD [13]. Thus, considering the yield of the 2,3-BD, sucrose as a cheap carbon source was chosen for the γ-PGA and 2,3-BD co-production.…”

“…In recent years, microbial fermentation for 2,3-BD production has been reported in a series of bacteria species, such as Klebsiella [7], Enterobacter [8], Paenibacillus [9], and Bacillus [10][11][12]. B. subtilis is a GRAS (generally regarded as safe) strain and also is considered to be a promising microbe for the production of 2,3-BD [13,14].…”

A Convenient Fluorescence-Based Assay for the Detection of Sucrose Transport and the Introduction of a Sucrose Transporter from Potato into Clostridium Strains

“…The maltose transporter was genetically characterized in Thermotoga maritima , and genetic engineering of this transporter improved the fermentation efficiency of this bacterium [12,13]. Introduction of a sucrose permease from Escherichia coli and a sucrose phosphorylase from Streptococcus mutans into Bacillus subtilis resulted in a recombinant strain that showed remarkable enhancement of 2,3-butanediol production compared with the control strain [14]. Overexpression of an endogenous sucrose utilization system, the phosphoenolpyruvate-phosphotransferase system (PEP-PTS) transport protein for sucrose and the related sucrose hydrolase enzymes elevated sucrose consumption and enhanced acetone-butanol-ethanol (ABE) production in Clostridium saccharoperbutylacetonicum N1-4 [15].…”

Efficient 2,3-butanediol production from whey powder using metabolically engineered Klebsiella oxytoca

“…One common method for BDO synthesis is performed under harsh conditions (160-220 °C, 50 bar) with a C 4 hydrocarbon fraction of cracked gases as the substrate [9,10]. However, due to shortage of fossil fuels and increasing global environmental concerns, green production of BDO through microbial fermentation is desirable [11][12][13][14][15][16]. Renewable resources such as rice waste biomass,…”