Purification and Properties of Acetylacetoin Synthase from<i>Bacillus</i>sp. YUF-4

Ui, Sadaharu; Hosaka, Takeshi; Mizutani, Kimiya; Mimura, Akio

doi:10.1271/bbb.62.795

Cited by 13 publications

(7 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An alternate route to 2,3-butanediol from acetoin via diacetyl and acetylacetoin has also been suggested in which diacetyl is acetylated and reduced to yield acetylbutanediol which is then hydrolyzed to 2,3-butanediol and acetate [23]. We cannot rule out that.…”

Section: Resultsmentioning

confidence: 96%

Novel monosaccharide fermentation products in Caldicellulosiruptor saccharolyticus identified using NMR spectroscopy

Isern

Xue

Rao

et al. 2013

Biotechnol Biofuels

View full text Add to dashboard Cite

BackgroundCaldicellulosiruptor saccharolyticus is a thermophilic, Gram-positive, non-spore forming, strictly anaerobic bacterium of interest in potential industrial applications, including the production of biofuels such as hydrogen or ethanol from lignocellulosic biomass through fermentation. High-resolution, solution-state nuclear magnetic resonance (NMR) spectroscopy is a useful method for the identification and quantification of metabolites that result from growth on different substrates. NMR allows facile resolution of isomeric (identical mass) constituents and does not destroy the sample.ResultsProfiles of metabolites produced by the thermophilic cellulose-degrading bacterium Caldicellulosiruptor saccharolyticus DSM 8903 strain following growth on different monosaccharides (D-glucose, D-mannose, L-arabinose, D-arabinose, D-xylose, L-fucose, and D-fucose) as carbon sources revealed several unexpected fermentation products, suggesting novel metabolic capacities and unexplored metabolic pathways in this organism. Both 1H and 13C nuclear magnetic resonance (NMR) spectroscopy were used to determine intracellular and extracellular metabolite profiles. One dimensional 1H NMR spectral analysis was performed by curve fitting against spectral libraries provided in the Chenomx software; 2-D homonuclear and heteronuclear NMR experiments were conducted to further reduce uncertainties due to unassigned, overlapping, or poorly-resolved peaks. In addition to expected metabolites such as acetate, lactate, glycerol, and ethanol, several novel fermentation products were identified: ethylene glycol (from growth on D-arabinose), acetoin and 2,3-butanediol (from growth on D-glucose, L-arabinose, and D-xylose), and hydroxyacetone (from growth on D-mannose, L-arabinose, and D-xylose). Production of ethylene glycol from D-arabinose was particularly notable, with around 10% of the substrate carbon converted into this uncommon fermentation product.ConclusionsThe present research shows that C. saccharolyticus, already of substantial interest due to its capability for biological ethanol and hydrogen production, has further metabolic potential for production of higher molecular weight compounds, such as acetoin and 2,3-butanediol, as well as hydroxyacetone and the uncommon fermentation product ethylene glycol. In addition, application of nuclear magnetic resonance (NMR) spectroscopy facilitates identification of novel metabolites, which is instrumental for production of desirable bioproducts from biomass through microbial fermentation.

show abstract

Section: Resultsmentioning

confidence: 96%

Novel monosaccharide fermentation products in Caldicellulosiruptor saccharolyticus identified using NMR spectroscopy

Isern

Xue

Rao

et al. 2013

Biotechnol Biofuels

View full text Add to dashboard Cite

show abstract

“…Information on the gene sequence and protein structure is not available for AAS and AAR of B. licheniformis but only for enzymes from bacteria of the same genus [27,31,32]. We recently found, however, that the combined use of AAS and AAR from B. licheniformis allows to prepare optically pure ␣-alkyl-␣,␤-dihydroxy ketones from 1,2-dione substrates by a two-step coupling/reduction sequence [26].…”

Section: Resultsmentioning

confidence: 99%

“…[27]). A detailed description of the AAS purification study has been the object of a PhD thesis from our group (see Ref.…”

Section: Resultsmentioning

confidence: 99%

“…Partially purified [27,28] AAS (1 U) was added to a solution of 2,3-octanedione 1 (20 mg, 0.14 mmol), ThDP (1.5 mg, 3.3 mol), and MgSO 4 (1.5 mg, 12.5 mol) in 50 mM phosphate buffer at pH 6.5 (10 mL) containing EDTA (0.1 mM) and 2-mercaptoethanol (1 mM). The reaction was gently shaken at 30 • C for 20 h and then heated (80 • C, 20 min) to denature proteins.…”

Section: Homo-coupling Of 23-octanedione (1) With Partially Purifiedmentioning

confidence: 99%

See 1 more Smart Citation

An enzymatic approach to the synthesis of optically pure (3R)- and (3S)-enantiomers of green tea flavor compound 3-hydroxy-3-methylnonane-2,4-dione

Bortolini

Giovannini

Maietti

et al. 2013

Journal of Molecular Catalysis B: Enzymatic

View full text Add to dashboard Cite

“…Some authors have described AAS as the first enzyme of apathway known as the "2,3-butanediol cycle", in which AAS is supposed to catalyze the ThDP-dependent condensation of two molecules of 2,3butanedione (1a)y ielding acetylacetoin (2a)a nd acetic acid through the formation of the (hydroxyethyl)thiamine diphosphate intermediate I (Scheme 1a). [13] Recently,h owever,t he "2,3-butanediol cycle" has been brought into question [14] and the currently most accepted mechanism for the bacterial degradation of acetoin relies on the action of the acetoin dehydrogenase enzyme system (AoDH ES). [14,15] Thef irst enzyme of this multienzymatic system, named acetoin:dichlorophenolindophenol oxidoreductase (Ao:DCPIP OR), catalyzes the ThDP-dependent oxidative cleavage of acetoin (3)l eading to acetaldehyde with transfer of the activated aldehyde to the lipoamide cofactor of the second enzyme of the system (Scheme 1b).…”

mentioning

confidence: 99%

Enzymatic Chemoselective Aldehyde–Ketone Cross‐Couplings through the Polarity Reversal of Methylacetoin

et al. 2015

View full text Add to dashboard Cite

The thiamine diphosphate (ThDP) dependent enzyme acetoin:dichlorophenolindophenol oxidoreductase (Ao:DCPIP OR) from Bacillus licheniformis was cloned and overexpressed in Escherichia coli. The recombinant enzyme shared close similarities with the acetylacetoin synthase (AAS) partially purified from Bacillus licheniformis suggesting that they could be the same enzyme. The product scope of the recombinant Ao:DCPIP OR was expanded to chiral tertiary α‐hydroxy ketones through the rare aldehyde–ketone cross‐carboligation reaction. Unprecedented is the use of methylacetoin as the acetyl anion donor in combination with a range of strongly to weakly activated ketones. In some cases, Ao:DCPIP OR produced the desired tertiary alcohols with stereochemistry opposite to that obtained with other ThDP‐dependent enzymes. The combination of methylacetoin as acyl anion synthon and novel ThDP‐dependent enzymes considerably expands the available range of CC bond formations in asymmetric synthesis.

show abstract

Purification and Properties of Acetylacetoin Synthase fromBacillussp. YUF-4

Cited by 13 publications

References 9 publications

Novel monosaccharide fermentation products in Caldicellulosiruptor saccharolyticus identified using NMR spectroscopy

Novel monosaccharide fermentation products in Caldicellulosiruptor saccharolyticus identified using NMR spectroscopy

An enzymatic approach to the synthesis of optically pure (3R)- and (3S)-enantiomers of green tea flavor compound 3-hydroxy-3-methylnonane-2,4-dione

Enzymatic Chemoselective Aldehyde–Ketone Cross‐Couplings through the Polarity Reversal of Methylacetoin

Contact Info

Product

Resources

About