Production of methyl ethyl ketone from biomass using a hybrid biochemical/catalytic approach

The recent demand for sustainable routes to fuels and chemicals has led to an increased amount of research in conversion of natural resources. A potential approach for conversion of biomass to fuels and chemicals is to combine biochemical and chemical processes. This research used microbial fermentation to produce 2,3-butanediol, which was then converted to methyl ethyl ketone by dehydration over a solid acid catalyst. The fermentation process was performed using the bacteria Klebsiella oxytoca (K.O). 2,3-butanediol then dehydrated to form methyl ethyl ketone on a solid acid catalyst, the proton form of ZSM-5, and heat. The goal was to determine the reaction kinetics of 2,3-butanediol dehydration over ZSM-5, and to demonstrate the hybrid biochemical/thermochemical approach for synthesizing chemicals from biomass. It was found that ZSM-5 produced methyl ethyl ketone with high selectivity (greater than 90%), and could convert fermentative 2,3-butanediol to methyl ethyl ketone. The reaction order of 2,3-butanediol dehydration was found to be slightly large than one, and an activation energy of 32.3 kJ/mol was measured

Production of 2,3-butanediol in Saccharomyces cerevisiae by in silico aided metabolic engineering

BACKGROUND: 2,3-Butanediol is a chemical compound of increasing interest due to its wide applications. It can be synthesized via mixed acid fermentation of pathogenic bacteria such as Enterobacter aerogenes and Klebsiella oxytoca. The non-pathogenic Saccharomyces cerevisiae possesses three different 2,3-butanediol biosynthetic pathways, but produces minute amount of 2,3-butanediol. Hence, we attempted to engineer S. cerevisiae strain to enhance 2,3-butanediol production. RESULTS: We first identified gene deletion strategy by performing in silico genome-scale metabolic analysis. Based on the best in silico strategy, in which disruption of alcohol dehydrogenase (ADH) pathway is required, we then constructed gene deletion mutant strains and performed batch cultivation of the strains. Deletion of three ADH genes, ADH1, ADH3 and ADH5, increased 2,3-butanediol production by 55-fold under microaerobic condition. However, overproduction of glycerol was observed in this triple deletion strain. Additional rational design to reduce glycerol production by GPD2 deletion altered the carbon fluxes back to ethanol and significantly reduced 2,3-butanediol production. Deletion of ALD6 reduced acetate production in strains lacking major ADH isozymes, but it did not favor 2,3-butanediol production. Finally, we introduced 2,3-butanediol biosynthetic pathway from Bacillus subtilis and E. aerogenes to the engineered strain and successfully increased titer and yield. Highest 2,3-butanediol titer (2.29 g·l(-1)) and yield (0.113 g·g(-1)) were achieved by Δadh1 Δadh3 Δadh5 strain under anaerobic condition. CONCLUSIONS: With the aid of in silico metabolic engineering, we have successfully designed and constructed S. cerevisiae strains with improved 2,3-butanediol production

2,3-butanediol in experimental myocardial ischaemia in pigs

To investigate the role of 2,3-butanediol in myocardial ischaemia we analysed this compound in pig's myocardium and blood. Ischaemia was induced by ligation of a coronary artery. In the first study we found significantly higher levels of 2,3-butanediol in the homogenate of ischaemic myocardium than in non-ischaemic myocardium. The lactate concentration was also significantly elevated. In the second study, where ischaemia was similarly induced, and where reperfusion was achieved by re-opening the ligated coronary artery after 20 min, 2,3-butanediol in peripheral blood was found to increase significantly. In the pigs in which the coronary artery was not re-opened, the 2,3-butanediol level in peripheral blood was unchanged. We conclude that in pigs' anaerobic myocardia accumulation of 2,3-butanediol occurs; if the myocardium is reperfused this metabolite also appears in the bloo

Production of 2-butanol through meso-2,3-butanediol consumption in lactic acid bacteria

2-Butanol has been an issue of industries in many areas, for example, biofuel production (as an advanced alternate fuel), fermented beverages, and food (as taste-altering component). Thus, its source of production, the biological pathway, and the enzymes involved are of high interest. In this study, 42 different isolates of lactic acid bacteria from nine different species were screened for their capability to consume meso-2,3-butanediol and produce 2-butanol. Lactobacillus brevis was the only species that showed any production of 2-butanol. Five of ten tested isolates of L.brevis were able to convert meso-2,3-butanediol to 2-butanol in a synthetic medium (SM2). However, none of them showed the same capability in a complex medium such as MRS indicating that the ability to produce 2-butanol is subject to some kind of repression mechanism. Furthermore, by evaluating the performance of the enzymes required to convert meso-2,3-butanediol to 2-butanol, that is, the secondary alcohol dehydrogenase and the diol dehydratase, it was shown that the latter needed the presence of a substrate to be expressed

Kinetics Modeling of the Heterogeneously Catalyzed Esterification of 2,3-Butanediol with Acetic Acid

Esterification of 2,3-butanediol using a homogeneous catalyst was studied in 1945 under the Synthetic Rubber Program. The aim of this paper is to show that this reaction can be carried out with a heterogeneous catalyst, which presents many advantages such as the possibility of separation and recycling. The kinetic behavior of heterogeneous esterification of 2,3-butanediol with acetic acid over an ion-exchange resin Amberlyst 36 was investigated in a batch reactor. The experiments were conducted with different amounts of catalyst (from 1.1% to 4.4%, relatively to the initial molar quantity of 2,3-butanediol), with molar ratios of reactants (acetic acid to 2,3-butanediol) varying from 2 to 12, and at temperatures from 50 to 110 °C. Our experimental system was able to acquire kinetic data, even during the first minutes of the reaction. A simple model based on reliable hypotheses was built and the experimental data were used to determine all the kinetic and thermodynamic constants of the reaction. A single set of parameters is able to represent correctly the evolution of all different species present during the esterification reaction in function of time

Multivariate statistical analysis for the identification of potential seafood spoilage indicators

Volatile organic compounds (VOCs) characterize the spoilage of seafood packaged under modified atmospheres (MAs) and could thus be used for quality monitoring. However, the VOC profile typically contains numerous multicollinear compounds and depends on the product and storage conditions. Identification of potential spoilage indicators thus calls for multivariate statistics. The aim of the present study was to define suitable statistical methods for this purpose (exploratory analysis) and to consequently characterize the spoilage of brown shrimp (Crangon crangon) and Atlantic cod (Gadus morhua) stored under different conditions (selective analysis). Hierarchical cluster analysis (HCA), principal components analysis (PCA) and partial least squares regression analysis (PLS) were applied as exploratory techniques (brown shrimp, 4 °C, 50%CO2/50%N2) and PLS was further selected for spoilage marker identification. Evolution of acetic acid, 2,3-butanediol, isobutyl alcohol, 3-methyl-1-butanol, dimethyl sulfide, ethyl acetate and trimethylamine was frequently in correspondence with changes in the microbiological quality or sensory rejection. Analysis of these VOCs could thus enhance the detection of seafood spoilage and the development of intelligent packaging technologies.acceptedVersionPeer reviewe

Genome Sequence of Klebsiella oxytoca M5al, a Promising Strain for Nitrogen Fixation and Chemical Production

Klebsiella oxytoca is an important microorganism for nitrogen fixation and chemical production. Here, we report an annotated draft genome of K. oxytoca strain M5al that contains 5,256 protein-coding genes and 95 structural RNAs, which provides a genetic basis for a better understanding of the physiology of this species

Adsorption of small organic solutes from beet distillery condensates on reverse-osmosis membranes: Consequences on the process performances

International audienceReverse osmosis (RO) is an attractive process for the detoxification of distillery condensates before their recycling at the fermentation stage. However, transfer mechanisms through dense NF and RO membranes are still not well understood for the organic solutes to eliminate, and rejection results could be disappointing. This study aims at correlating the membrane and solute characteristics (polarity, surface charge, molecular weight, etc.) to the transfer results and further to the process performances. This was achieved through the study of the sorption isotherms of five target inhibitory compounds (acetic and butanoic acids, furfural, 2-phenethyl alcohol and 2,3-butanediol) on three commercial RO membranes (ESPA2, CPA2 and BW30), the compounds being alone as well as mixed in order to evaluate the competition effects. Results reveal that acetic acid and 2,3-butanediol develop few interactions with the membrane material while furfural and 2-phenethyl alcohol present strong sorption of Langmuir type. Extended Langmuir equation succeeded in accounting for the solutes' sorption in mixtures insofar as acetic acid is not considered as competitor since it follows a different transfer mechanism. Coupled with their molecular weight (MW), low rejections obtained for acetic acid and furfural at pilot-scale as well as high rejection results for 2,3-butanediol and 2-phenylethanol could be explained

Esters of (S)-1,2-propanediol and (R,R)-2,3-butanediol — Chiral Compounds Inducing Cholesteric Phases with a Helix Inversion ·

Mesogenic chiral esters of optically active (S)-1,2-propanediol and (R,R)-2,3-butanediol were synthesized. The compounds, added to a nematic phase induce cholesteric phases exhibiting a helix inversion with temperature variation. This effect is independent of the molecular structure of the nematic solvent. The inversion temperature varies only slightly with concentration but can be influenced by the mesogenic substituent