Regulation of the NADH pool and NADH/NADPH ratio redistributes acetoin and 2,3‐butanediol proportion in <i>Bacillus subtilis</i>

Teng, Bo; Zhang, Xian; Zhao, Xiaojing; Rao, Zhiming; Yang, Taowei; Yang, Shang‐Tian

doi:10.1002/biot.201400577

Cited by 50 publications

(36 citation statements)

References 32 publications

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“…We observed that as soon as glucose had been depleted, R-BDO was oxidized, and this probably provided the NADH needed for NOX to maintain a reduced intracellular environment and thereby prevent unnecessary oxidative stress. 2,3-BDO oxidation upon substrate exhaustion has also been observed in previous studies37.…”

Section: Discussionsupporting

confidence: 83%

Synthesis of (3R)-acetoin and 2,3-butanediol isomers by metabolically engineered Lactococcus lactis

Kandasamy

Liu

Dantoft

et al. 2016

Sci Rep

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The potential that lies in harnessing the chemical synthesis capabilities inherent in living organisms is immense. Here we demonstrate how the biosynthetic machinery of Lactococcus lactis, can be diverted to make (3R)-acetoin and the derived 2,3-butanediol isomers meso-(2,3)-butanediol (m-BDO) and (2R,3R)-butanediol (R-BDO). Efficient production of (3R)-acetoin was accomplished using a strain where the competing lactate, acetate and ethanol forming pathways had been blocked. By introducing different alcohol dehydrogenases into this strain, either EcBDH from Enterobacter cloacae or SadB from Achromobacter xylosooxidans, it was possible to achieve high-yield production of m-BDO or R-BDO respectively. To achieve biosustainable production of these chemicals from dairy waste, we transformed the above strains with the lactose plasmid pLP712. This enabled efficient production of (3R)-acetoin, m-BDO and R-BDO from processed whey waste, with titers of 27, 51, and 32 g/L respectively. The corresponding yields obtained were 0.42, 0.47 and 0.40 g/g lactose, which is 82%, 89%, and 76% of maximum theoretical yield respectively. These results clearly demonstrate that L. lactis is an excellent choice as a cell factory for transforming lactose containing dairy waste into value added chemicals.

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Section: Discussionsupporting

confidence: 83%

Synthesis of (3R)-acetoin and 2,3-butanediol isomers by metabolically engineered Lactococcus lactis

Kandasamy

Liu

Dantoft

et al. 2016

Sci Rep

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“…The gene gdh with the upstream region including Hpa II promoter was amplified using forward primer P 1 and reverse primer P 2 using pMA5‐ gdh as template . The amplified gene was first inserted into simple cloning vector pMD18T TaKaRa Co. (Dalian, China)) by T4 ligase enzyme under the control of Hpa II promoter, forming to pMD18T‐ Hpa II ‐gdh .…”

Section: Methodsmentioning

confidence: 99%

Optimized whole cell biocatalyst from acetoin to 2,3‐butanediol through coexpression of acetoin reductase with NADH regeneration systems in engineered Bacillus subtilis

Samuel

Teng

Zhang

et al. 2017

J of Chemical Tech & Biotech

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BACKGROUND: 2,3-Butanediol (2,3-BD) has a wide range of applications in chiral molecular synthesis, biofuel additives, and in food flavor additive manufacturing. Fermentation is a favorable method for 2,3-BD production. However, it requires much time and produces several NADH related byproducts which compete with 2,3-BD production. Bacillus subtilis has an excellent ability for 2,3-BD production by biocatalysis. However, its production is limited by low intracellular NADH and the reversible property of acetoin reductase (AR/2,3-BDH). The whole cell biocatalyst process with two different NADH regeneration systems was designed for efficient production of 2,3-BD in B. subtilis 168. RESULTS: Formate dehydrogenase and glucose dehydrogenase for NADH regeneration were successfully co-expressed with acetoin reductase in B. subtilis 168. After optimization of biocatalyst bioconversion conditions, B. subtilis 168/pMA5-bdhA-HpaII-fdh yielded 74.5 g L − 1 of 2, 3-BD with 9.3 g L −1 h −1 productivity by fed batch and 115.4 g of 2,3-BD was achieved using same batch bacterium by three repeated batch bioconversions. On the other hand, 63.7 g L −1 of 2, 3-BD was produced with 7.92 g L −1 h −1 productivity by B. subtilis 168/pMA5-bdhA-HpaII-gdh. To our knowledge, the volume productivity obtained here is the highest ever reported for biocatalysis. CONCLUSION: A higher productivity of 2,3-BD from acetoin was achieved by whole cell biocatalysis with NADH regeneration systems in B. subtilis 168. This approach can be applied for NADH related bio-based chemicals production to improve titer, yield and productivity.

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“…Acetoin, which occurs naturally in corn, grapes, cocoa, apples, butter, coffee, and beer, is a famous flavor agent. During production of acetoin, several NADH‐dependent byproducts always appear, especially 2,3‐butanediol (2,3‐BDL) . Some strategies, e.g.…”

Section: Introductionmentioning

confidence: 99%

Artificial consortium that produces riboflavin regulates distribution of acetoin and 2,3‐butanediol by Paenibacillus polymyxa CJX518

Liu

Chen

et al. 2017

Engineering in Life Sciences

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The introduction of an NADH/NAD+ regeneration system can regulate the distribution between acetoin and 2,3‐butanediol. NADH regeneration can also enhance butanol production in coculture fermentation. In this work, a novel artificial consortium of Paenibacillus polymyxa CJX518 and recombinant Escherichia coli LS02T that produces riboflavin (VB2) was used to regulate the NADH/NAD+ ratio and, consequently, the distribution of acetoin and 2,3‐butanediol by P. polymyxa. Compared with a pure culture of P. polymyxa, the level of acetoin was increased 76.7% in the P. polymyxa and recombinant E. coli coculture. Meanwhile, the maximum production and yield of acetoin in an artificial consortium with fed‐batch fermentation were 57.2 g/L and 0.4 g/g glucose, respectively. Additionally, the VB2 production of recombinant E. coli could maintain a relatively low NADH/NAD+ ratio by changing NADH dehydrogenase activity. It was also found that 2,3‐butanediol dehydrogenase activity was enhanced and improved acetoin production by the addition of exogenous VB2 or by being in the artificial consortium that produces VB2. These results illustrate that the coculture of P. polymyxa and recombinant E. coli has enormous potential to improve acetoin production. It was also a novel strategy to regulate the NADH/NAD+ ratio to improve the acetoin production of P. polymyxa.

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Regulation of the NADH pool and NADH/NADPH ratio redistributes acetoin and 2,3‐butanediol proportion in Bacillus subtilis

Cited by 50 publications

References 32 publications

Synthesis of (3R)-acetoin and 2,3-butanediol isomers by metabolically engineered Lactococcus lactis

Synthesis of (3R)-acetoin and 2,3-butanediol isomers by metabolically engineered Lactococcus lactis

Optimized whole cell biocatalyst from acetoin to 2,3‐butanediol through coexpression of acetoin reductase with NADH regeneration systems in engineered Bacillus subtilis

Artificial consortium that produces riboflavin regulates distribution of acetoin and 2,3‐butanediol by Paenibacillus polymyxa CJX518

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