Metabolic engineering of Bacillus subtilis for enhanced production of acetoin

Wang, Meng; Fu, Jing; Zhang, Xueyu; Chen, Tao

doi:10.1007/s10529-012-0981-9

Cited by 54 publications

(33 citation statements)

References 13 publications

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“…The cells were resuspended in PBS buffer and then disrupted at 4°C with 1-s bursts of ultrasonic sound at 3-s intervals over a total period of 15 min. Disruption was confirmed by microscopy [27]. The lysates were centrifuged at 12,000 rpm for 10 min at 4°C, and the supernatants were then analyzed for XYL1 activity.…”

Section: Enzyme Activitymentioning

confidence: 99%

Production of Xylitol from d-Xylose by Overexpression of Xylose Reductase in Osmotolerant Yeast Candida glycerinogenes WL2002-5

Zhang

Zong

Zhuge

et al. 2015

Appl Biochem Biotechnol

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Efficient bioconversion of D-xylose into various biochemicals is critical for the developing lignocelluloses application. In this study, we compared D-xylose utilization in Candida glycerinogenes WL2002-5 transformants expressing xylose reductase (XYL1) in D-xylose metabolism. C. glycerinogenes WL2002-5 expressing XYL1 from Schefferomyces stipitis can produce xylitol. Xylitol production by the recombinant strains was evaluated using a xylitol fermentation medium with glucose as a co-substrate. As glucose was found to be an insufficient co-substrate, various carbon sources were screened for efficient cofactor regeneration, and glycerol was found to be the best co-substrate. The effects of glycerol on the xylitol production rate by a xylose reductase gene (XYL1)-overexpressed mutant of C. glycerinogenes WL2002-5 were investigated. The XYL1-overexpressed mutant produced xylitol from D-xylose using glycerol as a co-substrate for cell growth and NAD (P) H regeneration: 100 g/L D-xylose was completely converted into xylitol when at least 20 g/L glycerol was used as a co-substrate. XYL1 overexpressed mutant grown on glycerol as co-substrate accumulated 2.1-fold increased xylitol concentration over those cells grown on glucose as co-substrate. XYL1 overexpressed mutant produced xylitol with a volumetric productivity of 0.83 g/L/h, and a xylitol yield of 98 % xylose. Recombinant yeast strains obtained in this study are promising candidates for xylitol production. This is the first report of XYL1 gene overexpression of C. glycerinogenes WL2002-5 for enhancing the efficiency of xylitol production.

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Section: Enzyme Activitymentioning

confidence: 99%

Production of Xylitol from d-Xylose by Overexpression of Xylose Reductase in Osmotolerant Yeast Candida glycerinogenes WL2002-5

Zhang

Zong

Zhuge

et al. 2015

Appl Biochem Biotechnol

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“…This substance is widely used in the food, cosmetics, perfume, pharmacy, and chemistry industries. It is one of thirty basic chemical components that the U.S. Department of Energy has prioritized for development and utilization [3]. [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, the conventional methods include ion-exchange (IE-) and reversed-phase (RP-) HPLC [3][4][5][11][12][13]. The GC and GC-MS-MS methods are the most commonly used, but they have certain disadvantages, such as the complexity of sample preparation and the high cost of analysis.…”

Section: Introductionmentioning

confidence: 99%

HPLC and UPLC Analyses of Acetoin in Bacterial Culture Fluid

Федорова¹,

Kivero²,

Geraskina³

et al. 2015

Acta Chromatographica

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Summary.A simple and rapid ultra-performance liquid chromatographic (UPLC) method for analyzing acetoin in bacterial culture fluid was developed and validated for the first time. The samples were separated using an Acquity BEH C18 column (2.1 mm × 100 mm, 1.7 μm particle size) and isocratic elution with 30 mM phosphoric acid-1% acetonitrile as the mobile phase. A photodiode array detector (PDA) was used. The run time was 6 min, and the detection limit was 2.11 × 10 −4 mg mL −1 . The UPLC method was compared with high-pressure liquid chromatography (HPLC) for acetoin analysis. The proposed UPLC method is highly sensitive and was successfully applied to the analysis of acetoin in bacterial culture fluid.

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“…Widely used in food and beverage industry, AC also serves as a platform compound in many other industries [1]. It is one of the 30 platform chemicals that are given priority to their development and utilization by the US Department of Energy [2].…”

Section: Introductionmentioning

confidence: 99%

Efficient Whole-Cell Biocatalyst for Acetoin Production with NAD+ Regeneration System through Homologous Co-Expression of 2,3-Butanediol Dehydrogenase and NADH Oxidase in Engineered Bacillus subtilis

et al. 2014

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Acetoin (3-hydroxy-2-butanone), an extensively-used food spice and bio-based platform chemical, is usually produced by chemical synthesis methods. With increasingly requirement of food security and environmental protection, bio-fermentation of acetoin by microorganisms has a great promising market. However, through metabolic engineering strategies, the mixed acid-butanediol fermentation metabolizes a certain portion of substrate to the by-products of organic acids such as lactic acid and acetic acid, which causes energy cost and increases the difficulty of product purification in downstream processes. In this work, due to the high efficiency of enzymatic reaction and excellent selectivity, a strategy for efficiently converting 2,3-butandiol to acetoin using whole-cell biocatalyst by engineered Bacillus subtilis is proposed. In this process, NAD+ plays a significant role on 2,3-butanediol and acetoin distribution, so the NADH oxidase and 2,3-butanediol dehydrogenase both from B. subtilis are co-expressed in B. subtilis 168 to construct an NAD+ regeneration system, which forces dramatic decrease of the intracellular NADH concentration (1.6 fold) and NADH/NAD+ ratio (2.2 fold). By optimization of the enzymatic reaction and applying repeated batch conversion, the whole-cell biocatalyst efficiently produced 91.8 g/L acetoin with a productivity of 2.30 g/(L·h), which was the highest record ever reported by biocatalysis. This work indicated that manipulation of the intracellular cofactor levels was more effective than the strategy of enhancing enzyme activity, and the bioprocess for NAD+ regeneration may also be a useful way for improving the productivity of NAD+-dependent chemistry-based products.

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Metabolic engineering of Bacillus subtilis for enhanced production of acetoin

Cited by 54 publications

References 13 publications

Production of Xylitol from d-Xylose by Overexpression of Xylose Reductase in Osmotolerant Yeast Candida glycerinogenes WL2002-5

Production of Xylitol from d-Xylose by Overexpression of Xylose Reductase in Osmotolerant Yeast Candida glycerinogenes WL2002-5

HPLC and UPLC Analyses of Acetoin in Bacterial Culture Fluid

Efficient Whole-Cell Biocatalyst for Acetoin Production with NAD+ Regeneration System through Homologous Co-Expression of 2,3-Butanediol Dehydrogenase and NADH Oxidase in Engineered Bacillus subtilis

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