Showing papers on "2,3-Butanediol published in 2022"

Low dose 1,3-butanediol reverses age-associated vascular dysfunction independent of ketone body β-hydroxybutyrate.

Abstract: With an aging global population, identifying novel therapeutics are necessary to increase longevity and decrease the deterioration of essential end organs such as the vasculature. Secondary alcohol, 1,3-butanediol (1,3-BD), is commonly administered to stimulate the biosynthesis of the most abundant ketone body β-hydroxybutyrate (βHB), in lieu of nutrient deprivation. However, supra-pharmacological concentrations of 1,3-BD are necessary to significantly increase systemic βHB, and 1,3-BD per se can cause vasodilation at nanomolar concentrations. Therefore, we hypothesized that 1,3-BD could be a novel anti-aging therapeutic, independent of βHB biosynthesis. To test this hypothesis, we administered a low dose (5%) 1,3-BD to young and old Wistar-Kyoto (WKY) rats via drinking water for four weeks and measured indices of vascular function and metabolism post-treatment. We observed that low dose 1,3-BD was sufficient to reverse age-associated endothelial-dependent and -independent dysfunction, and this was not associated with increased βHB bioavailability. Further analysis of the direct vasodilator mechanisms of 1,3-BD revealed that it is predominantly an endothelium-dependent vasodilator through activation of potassium channels and nitric oxide synthase. In summary, we report that 1,3-BD, at a concentration that does not stimulate βHB biosynthesis, could be a nutraceutical that can reverse the age-associated decline in vascular function. These results emphasize that 1,3-BD has multiple, concentration-dependent mechanisms of action. Therefore, we suggest alternative approaches to study the physiological and cardiovascular effects of βHB.

Non-Sterilized Fermentation of 2,3-Butanediol with Seawater by Metabolic Engineered Fast-Growing Vibrio natriegens

Abstract: Sustainable and environment-friendly microbial fermentation processes have been developed to produce numerous chemicals. However, the high energy input required for sterilization and substantial fresh water consumption restrict the economic feasibility of traditional fermentation processes. To address these problems, Vibrio natriegens, a promising microbial chassis with low nutritional requirements, high salt tolerance and rapid growth rate can be selected as the host for chemical production. In this study, V. natriegens was metabolic engineered to produce 2,3-butanediol (2,3-BD), an important platform chemical, through non-sterilized fermentation with seawater-based minimal medium after expressing a 2,3-BD synthesis cluster and deleting two byproduct encoding genes. Under optimized fermentative conditions, 41.27 g/L 2,3-BD was produced with a productivity of 3.44 g/L/h and a yield of 0.39 g/g glucose by recombinant strain V. natriegensΔfrdAΔldhA-pETRABC. This study confirmed the feasibility of non-sterilized fermentation using seawater to replace freshwater and other valuable chemicals may also be produced through metabolic engineering of the emerging synthetic biology chassis V. natriegens.

Elicitation of (E)-2-Hexenal and 2,3-Butanediol on the Bioactive Compounds in Adventitious Roots of Astragalus membranaceus var. mongholicus.

Abstract: This study aimed to investigate the elicitation of volatile organic compounds (E)-2-hexenal and 2,3-butanediol on bioactive metabolites in Astragalus membranaceus var. mongholicus adventitious root cultures by adding them into the medium. The experiment was performed for 72 h and the roots were dynamically sampled for quantification of representative astragaloside IV, calycosin-7-O-β-d-glucoside (CG), ononin, and the gene expression. Compared with the controls, the combination of 2,3-butanediol and (E)-2-hexenal advanced the peak accumulation of astragaloside IV and was the most effective, but their individual application delayed it. Meanwhile, 2,3-butanediol and (E)-2-hexenal had no obviously promoting effect on the production of CG and ononin but chronologically changed their accumulation patterns. The underlying mechanism was uncovered by the correlation analysis between the metabolites and the gene expression, as did the identification of the target genes. Collectively, 2,3-butanediol and (E)-2-hexenal were important cues shaping the production of bioactive products in the herbal plant.

Metabolic engineering of Corynebacterium glutamicum for efficient production of optically pure (2R,3R)-2,3-butanediol

Abstract: 2,3-butanediol is an important platform compound which has a wide range of applications, involving in medicine, chemical industry, food and other fields. Especially the optically pure (2R,3R)-2,3-butanediol can be employed as an antifreeze agent and as the precursor for producing chiral compounds. However, some (2R,3R)-2,3-butanediol overproducing strains are pathogenic such as Enterobacter cloacae and Klebsiella oxytoca.In this study, a (3R)-acetoin overproducing C. glutamicum strain, CGS9, was engineered to produce optically pure (2R,3R)-2,3-butanediol efficiently. Firstly, the gene bdhA from B. subtilis 168 was integrated into strain CGS9 and its expression level was further enhanced by using a strong promoter Psod and ribosome binding site (RBS) with high translation initiation rate, and the (2R,3R)-2,3-butanediol titer of the resulting strain was increased by 33.9%. Then the transhydrogenase gene udhA from E. coli was expressed to provide more NADH for 2,3-butanediol synthesis, which reduced the accumulation of the main byproduct acetoin by 57.2%. Next, a mutant atpG was integrated into strain CGK3, which increased the glucose consumption rate by 10.5% and the 2,3-butanediol productivity by 10.9% in shake-flask fermentation. Through fermentation engineering, the most promising strain CGK4 produced a titer of 144.9 g/L (2R,3R)-2,3-butanediol with a yield of 0.429 g/g glucose and a productivity of 1.10 g/L/h in fed-batch fermentation. The optical purity of the resulting (2R,3R)-2,3-butanediol surpassed 98%.To the best of our knowledge, this is the highest titer of optically pure (2R,3R)-2,3-butanediol achieved by GRAS strains, and the result has demonstrated that C. glutamicum is a competitive candidate for (2R,3R)-2,3-butanediol production.

Production of Different Biochemicals by Paenibacillus polymyxa DSM 742 From Pretreated Brewers’ Spent Grains

Abstract: Brewers’ spent grains (BSG) are a by-product of the brewing industry that is mainly used as feedstock; otherwise, it has to be disposed according to regulations. Due to the high content of glucose and xylose, after pretreatment and hydrolysis, it can be used as a main carbohydrate source for cultivation of microorganisms for production of biofuels or biochemicals like 2,3-butanediol or lactate. 2,3-Butanediol has applications in the pharmaceutical or chemical industry as a precursor for varnishes and paints or in the food industry as an aroma compound. So far, Klebsiella pneumoniae, Serratia marcescens, Clostridium sp., and Enterobacter aerogenes are being used and investigated in different bioprocesses aimed at the production of 2,3-butanediol. The main drawback is bacterial pathogenicity which complicates all production steps in laboratory, pilot, and industrial scales. In our study, a gram-positive GRAS bacterium Paenibacillus polymyxa DSM 742 was used for the production of 2,3-butanediol. Since this strain is very poorly described in literature, bacterium cultivation was performed in media with different glucose and/or xylose concentration ranges. The highest 2,3-butanediol concentration of 18.61 g l–1 was achieved in medium with 70 g l–1 of glucose during 40 h of fermentation. In contrast, during bacterium cultivation in xylose containing medium there was no significant 2,3-butanediol production. In the next stage, BSG hydrolysates were used for bacterial cultivation. P. polymyxa DSM 742 cultivated in the liquid phase of pretreated BSG produced very low 2,3-butanediol and ethanol concentrations. Therefore, this BSG hydrolysate has to be detoxified in order to remove bacterial growth inhibitors. After detoxification, bacterium cultivation resulted in 30 g l–1 of lactate, while production of 2,3-butanediol was negligible. The solid phase of pretreated BSG was also used for bacterium cultivation after its hydrolysis by commercial enzymes. In these cultivations, P. polymyxa DSM 742 produced 9.8 g l–1 of 2,3-butanediol and 3.93 g l–1 of ethanol. On the basis of the obtained results, it can be concluded that different experimental setups give the possibility of directing the metabolism of P. polymyxa DSM 742 toward the production of either 2,3-butanediol and ethanol or lactate.

Biosynthesis of 1,3‐propanodiol and 2,3‐butanodiol from residual glycerol in continuous cell‐immobilized Klebsiella pneumoniae bioreactors

Abstract: In recent years, residual glycerol from biodiesel synthesis made this chemical a cheap, readily available carbon source to bioprocess, which is also a form to reduce costs in the fuel industry. We propose and describe a bioprocess using fluidized and packed‐bed continuous bioreactors to convert this residual glycerol into value‐added products such as 1,3‐propanediol (1,3‐PD) and 2,3‐butanediol (2,3‐BD), largely used in the chemical industry. The bacterium Klebsiella pneumoniae BLh‐1, strain isolated by us, was immobilized in the permeable support of polyvinyl alcohol (LentiKats®). After testing different dilution rates (D) for all bioreactor configurations, the best obtained productivities of 1,3‐PD was 8.69 g L−1 h−1 at a D = 0.45 h−1, and 2.99 g L−1 h−1 at a D = 0.30 h−1 for 2,3‐BD, both in the packed‐bed configuration. In the fluidized‐bed reactor, the highest productivity values achieved were 4.48 and 1.16 g L−1 h−1 for 1,3‐PD and 2,3‐BD, respectively, both at D = 0.33 h−1. These results show the potential of setting up a bioprocess based on continuous cultures using immobilized K. pneumoniae BLh‐1 in PVA matrices in order to efficiently convert the abundant surplus of glycerol into commercially important chemicals such as 1,3‐PD and 2,3‐BD.

Development of an industrial yeast strain for efficient production of 2,3-butanediol

Abstract: As part of the transition from a fossil resources-based economy to a bio-based economy, the production of platform chemicals by microbial cell factories has gained strong interest. 2,3-butanediol (2,3-BDO) has various industrial applications, but its production by microbial fermentation poses multiple challenges. We have engineered the bacterial 2,3-BDO synthesis pathway, composed of AlsS, AlsD and BdhA, in a pdc-negative version of an industrial Saccharomyces cerevisiae yeast strain. The high concentration of glycerol caused by the excess NADH produced in the pathway from glucose to 2,3-BDO was eliminated by overexpression of NoxE and also in a novel way by combined overexpression of NDE1, encoding mitochondrial external NADH dehydrogenase, and AOX1, encoding a heterologous alternative oxidase expressed inside the mitochondria. This was combined with strong downregulation of GPD1 and deletion of GPD2, to minimize glycerol production while maintaining osmotolerance. The HGS50 strain produced a 2,3-BDO titer of 121.04 g/L from 250 g/L glucose, the highest ever reported in batch fermentation, with a productivity of 1.57 g/L.h (0.08 g/L.h per gCDW) and a yield of 0.48 g/g glucose or with 96% the closest to the maximum theoretical yield ever reported. Expression of Lactococcus lactis NoxE, encoding a water-forming NADH oxidase, combined with similar genetic modifications, as well as expression of Candida albicans STL1, also minimized glycerol production while maintaining high osmotolerance. The HGS37 strain produced 130.64 g/L 2,3-BDO from 280 g/L glucose, with productivity of 1.58 g/L.h (0.11 g/L.h per gCDW). Both strains reach combined performance criteria adequate for industrial implementation.

Biological Control Efficacy and Action Mechanism of Klebsiella pneumoniae JCK-2201 Producing Meso-2,3-Butanediol Against Tomato Bacterial Wilt

Abstract: Bacterial wilt caused by Ralstonia solanacearum is a fatal disease that affects the production of tomatoes and many other crops worldwide. As an effective strategy to manage bacterial wilt, biological control agents using plant growth-promoting rhizobacteria (PGPR) are being developed. In this study, we screened 2,3-butanediol (BDO)-producing PGPR to control tomato bacterial wilt and investigated the action mechanism of the disease control agent. Of the 943 strains isolated from soil, Klebsiella pneumoniae strain JCK-2201 produced the highest concentration of 2,3-BDO. The culture broth of K. pneumoniae JCK-2201 did not show any direct activity on R. solanacearum in vitro, but a 100-fold dilution effectively controlled tomato bacterial wilt with a control value of 77% in vivo. Fermentation utilizing K. pneumoniae JCK-2201 was optimized to produce 48 g/L of meso-2,3-BDO, which is 50% of the sucrose conversion efficiency. In addition, the control efficacy and mechanism of meso-2,3-BDO produced by JCK-2201 in tomato bacterial wilt were determined by comparative analysis with Bacillus licheniformis DSM13 producing meso-2,3-BDO and B. licheniformis DSM13 ΔalsS that did not produce 2,3-BDO, as the step of converting pyruvate to α-acetolactate was omitted. Tomato seedlings treated with the K. pneumoniae JCK-2201 (500-fold dilution) and B. licheniformis DSM13 (100-fold dilution) culture broth produced meso-2,3-BDO that significantly reduced R. solanacearum-induced disease severity with control values of 55% and 63%, respectively. The formulated meso-2,3-BDO 9% soluble concentrate (SL; 1,000-fold dilution) showed 87% control against tomato bacterial wilt in the field condition. Klebsiella pneumoniae JCK-2201 and B. licheniformis DSM13 treatment induced the expression of plant defense marker genes, such as LePR1, LePR2, LePR5, LePR3, and PI-II, in the salicylic acid and jasmonic acid signaling pathways at 4 days after inoculation. These results show that 2,3-BDO-producing bacteria and 2,3-BDO are potential biological control agents that act through induction of resistance for controlling tomato bacterial wilt.

Bioprocess development of 2, 3-butanediol production using agro-industrial residues

Abstract: The valorization of agricultural and industrial wastes for fuel and chemical production benefits environmental sustainability. 2, 3-Butanediol (2,3-BDO) is a value-added platform chemical covering many industrial applications. Since the global market is increasing drastically, production rates have to increase. In order to replace the current petroleum-based 2,3-BDO production, renewable feedstock's ability has been studied for the past few decades. This study aims to find an improved bioprocess for producing 2,3-BDO from agricultural and industrial residues, consequently resulting in a low CO2 emission bioprocess. For this, screening of 13 different biomass samples for hydrolyzable sugars has been done. Alkali pretreatment has been performed with the processed biomass and enzyme hydrolysis performed using commercial cellulase. Among all biomass hydrolysate oat hull and spruce bark biomass could produce the maximum amount of total reducing sugars. Later oat hull and spruce bark biomass with maximum hydrolyzable sugars have been selected for submerged fermentation studies using Enterobacter cloacae SG1. After fermentation, 37.59 and 26.74 g/L of 2,3-BDO was obtained with oat hull and spruce bark biomass, respectively. The compositional analysis of each step of biomass processing has been performed and changes in each component have been evaluated. The compositional analysis has revealed that biomass composition has changed significantly after pretreatment and hydrolysis leading to a remarkable release of sugars which can be utilized by bacteria for 2,3-BDO production. The results have been found to be promising, showing the potential of waste biomass residues as a low-cost raw material for 2,3-BDO production and thus a new lead in an efficient waste management approach for less CO2 emission.

Highly efficient production of 2,3-butanediol from xylose and glucose by newly isolated thermotolerant Cronobacter sakazakii

Abstract: 2,3-Butanediol (2,3-BD), a valuable compound used for chemicals, cosmetics, pesticides and pharmaceuticals, has been produced by various microbes. However, no high-temperature fermentation of the compound at high productivity has been reported.Thermotolerant xylose-utilizing microbes were isolated from 6 different districts in Laos and screened for a low accumulation of xylitol in a xylose medium at 37 ˚C. One isolate was found to produce 2,3-BD and identified by 16S rDNA sequencing. The 2,3-BD fermentation capacity was investigated at different temperatures using xylose and glucose as carbon sources, and the fermentation parameters were determined by a high-performance liquid chromatography system.By screening for a low accumulation of xylitol in a xylose medium, one isolate that accumulated almost no xylitol was obtained. Further analyses revealed that the isolate is Cronobacter sakazakii and that it has the ability to produce 2,3-BD at high temperatures. When xylose and glucose were used, this strain, named C. sakazakii OX-25, accumulated 2,3-BD in a short period before the complete consumption of these sugars and then appeared to convert 2,3-BD to acetoin. The optimum temperature of the 2,3-BD fermentation was 42 ˚C to 45 ˚C, and the maximum yield of 2,3-BD was 0.3 g/g at 12 h in 20 g/l xylose medium and 0.4 g/g at 6 h in 20 g/l glucose medium at 42 ˚C. The 2,3-BD productivity of the strain was higher than the 2,3-BD productivities of other non-genetically engineered microorganisms reported previously, and the highest productivity was 0.6 g/l·h and 1.2 g/l·h for xylose and glucose, respectively.Among thermotolerant microbes isolated in Laos, we discovered a strain, C. sakazakii OX-25, that can convert xylose and glucose to 2,3-BD with high efficiency and high productivity at high temperatures, suggesting that C. sakazakii OX-25 has the potential for industrial application to produce 2,3-BD as an important platform chemical.

Highly efficient production of 2,3-butanediol from xylose and glucose by newly isolated thermotolerant Cronobacter sakazakii

Abstract: 2,3-Butanediol (2,3-BD), a valuable compound used for chemicals, cosmetics, pesticides and pharmaceuticals, has been produced by various microbes. However, no high-temperature fermentation of the compound at high productivity has been reported.Thermotolerant xylose-utilizing microbes were isolated from 6 different districts in Laos and screened for a low accumulation of xylitol in a xylose medium at 37 ˚C. One isolate was found to produce 2,3-BD and identified by 16S rDNA sequencing. The 2,3-BD fermentation capacity was investigated at different temperatures using xylose and glucose as carbon sources, and the fermentation parameters were determined by a high-performance liquid chromatography system.By screening for a low accumulation of xylitol in a xylose medium, one isolate that accumulated almost no xylitol was obtained. Further analyses revealed that the isolate is Cronobacter sakazakii and that it has the ability to produce 2,3-BD at high temperatures. When xylose and glucose were used, this strain, named C. sakazakii OX-25, accumulated 2,3-BD in a short period before the complete consumption of these sugars and then appeared to convert 2,3-BD to acetoin. The optimum temperature of the 2,3-BD fermentation was 42 ˚C to 45 ˚C, and the maximum yield of 2,3-BD was 0.3 g/g at 12 h in 20 g/l xylose medium and 0.4 g/g at 6 h in 20 g/l glucose medium at 42 ˚C. The 2,3-BD productivity of the strain was higher than the 2,3-BD productivities of other non-genetically engineered microorganisms reported previously, and the highest productivity was 0.6 g/l·h and 1.2 g/l·h for xylose and glucose, respectively.Among thermotolerant microbes isolated in Laos, we discovered a strain, C. sakazakii OX-25, that can convert xylose and glucose to 2,3-BD with high efficiency and high productivity at high temperatures, suggesting that C. sakazakii OX-25 has the potential for industrial application to produce 2,3-BD as an important platform chemical.

Treatment of wild-type mice with 2,3-butanediol, a urinary biomarker of Fmo5 −/− mice, decreases plasma cholesterol and epididymal fat deposition

Abstract: We previously showed that Fmo5 −/− mice exhibit a lean phenotype and slower metabolic ageing. Their characteristics include lower plasma glucose and cholesterol, greater glucose tolerance and insulin sensitivity, and a reduction in age-related weight gain and whole-body fat deposition. In this paper, nuclear magnetic resonance (NMR) spectroscopy-based metabolite analyses of the urine of Fmo5 −/− and wild-type mice identified two isomers of 2,3-butanediol as discriminating urinary biomarkers of Fmo5 −/− mice. Antibiotic-treatment of Fmo5 −/− mice increased plasma cholesterol concentration and substantially reduced urinary excretion of 2,3-butanediol isomers, indicating that the gut microbiome contributed to the lower plasma cholesterol of Fmo5 −/− mice, and that 2,3-butanediol is microbially derived. Short- and long-term treatment of wild-type mice with a 2,3-butanediol isomer mix decreased plasma cholesterol and epididymal fat deposition but had no effect on plasma concentrations of glucose or insulin, or on body weight. In the case of long-term treatment, the effects were maintained after withdrawal of 2,3-butanediol. Short-, but not long-term treatment, also decreased plasma concentrations of triglycerides and non-esterified fatty acids. Fecal transplant from Fmo5 −/− to wild-type mice had no effect on plasma cholesterol, and 2,3-butanediol was not detected in the urine of recipient mice, suggesting that the microbiota of the large intestine was not the source of 2,3-butanediol. However, 2,3-butanediol was detected in the stomach of Fmo5 −/− mice, which was enriched for Lactobacillus genera, known to produce 2,3-butanediol. Our results indicate a microbial contribution to the phenotypic characteristic of Fmo5 −/− mice of decreased plasma cholesterol and identify 2,3-butanediol as a potential agent for lowering plasma cholesterol.

From Agricultural Wastes to Fermentation Nutrients: A Case Study of 2,3-Butanediol Production

Abstract: The goal of this study was to improve resource use efficiency in agricultural systems and agro-based industries, reduce wastes that go to landfills and incinerators, and consequently, improve the economics of 2,3-butanediol (2,3-BD) production. This study evaluated the feasibility of 2,3-BD production by replacing the mineral nutrients, and buffers with anaerobic digestate (ADE), poultry-litter (PLBC)- and forage-sorghum (FSBC)-derived biochars. Fermentation media formulations with ADE and 5–20 g/L PLBC or FSBC were evaluated for 2,3-BD production using Paenibacillus polymyxa as a biocatalyst. An optimized medium containing nutrients and buffers served as control. While 2,3-BD production in the ADE cultures was 0.5-fold of the maximum generated in the control cultures, 2,3-BD produced in the PLBC and FSBC cultures were ~1.3-fold more than the control (33.6 g/L). Cost analysis showed that ADE and biochar can replace mineral nutrients and buffers in the medium with the potential to make bio-based 2,3-BD production profitably feasible.

Biotechnological Production of Optically Pure 2,3-butanediol by Bacillus subtilis Based on Dissolved Oxygen Control Strategy

Abstract: 2,3-Butanediol (2,3-BD) is a promising platform chemical, produced from microbial cells. Oxygen availability is a crucial factor driving the formation and proportion of 2,3-BD and acetoin in 2,3-BD producing bacterial strains. In this study, the ability of B. subtills GD5 to produce 2,3-BD in optimized sucrose-based media was evaluated, by investigating the impact of carbon to nitrogen (C/N) ratio and the effectiveness of alternative low-cost nitrogen sources (corn steep liquor, soybean meal, and ammonium sulphate). Subsequently, different dissolved oxygen (DO) controlling regimes were assessed in batch bioreactor fermentations. The best fermentation outcomes were obtained with uncontrolled DO, achieving 5.88 g/L of optically pure (R,R)-2,3-BD (~100% purity), accompanied by a production yield of 0.43 g/g, and a productivity of 0.2 g/L/h. Additionally, the influence of the DO controlling regime on B. subtills key enzymes involved in the reverse activity of acetoin reductase was also monitored. A fed-batch process under the most suitable DO conditions was carried out to improve 2,3-BD production, achieving 42.31 g/L 2,3-BD with a production yield of 0.52 g/g. Thus, B. subtilis GD5 is a promising strain for the efficient production of pure chiral (R,R)-2,3-BD under uncontrolled DO conditions, using alternative low-cost nitrogen sources.

Control Properties of Sustainable Alternatives to Produce 2,3-Butanediol

Abstract: Current social needs, as well as political and environmental restrictions, have led to the generation of products within a framework of sustainability, and favoring production processes based on renewable raw materials. During the process design work, the role of the dynamic properties of a process, together with some other metrics, is not always considered. However, several studies have shown that there is a certain relationship between the dynamics and controllability of the process and its sustainability. In this work, the dynamic properties of different separation schemes for the purification of 2,3-BD from biomass fermentation are evaluated. The study allowed determining the best alternatives for the eventual implementation of these schemes, as well as the role of controllability concerning other sustainability indicators. As result, it was found that schemes that include a thermal coupling in their topological design allow a general improvement of all sustainability indicators. In addition, it was observed in this case study that process intensification contributes significantly to the fulfillment of various sustainability indicators. • It was obtained the control properties of separation alternatives to purify 2,3-Butanediol, the thermally coupled alternatives resulted as the most balanced alternatives • The sustainable evaluation allows considering a thermal coupling as a fundamental step in process design of a sustainable process. • The most intensified alternative did not result as the most sustainable alternative

Metabolic Engineering Interventions for Sustainable 2,3-Butanediol Production in Gas-Fermenting Clostridium autoethanogenum

Abstract: Envisioning value chains inspired by environmental sustainability and circularity in economic models is essential to counteract the alterations in the global natural carbon cycle induced by humans. Recycling carbon-based waste gas streams into chemicals by devising gas fermentation bioprocesses mediated by acetogens of the genus Clostridium is one component of the solution. ABSTRACT Gas fermentation provides a promising platform to turn low-cost and readily available single-carbon waste gases into commodity chemicals, such as 2,3-butanediol. Clostridium autoethanogenum is usually used as a robust and flexible chassis for gas fermentation. Here, we leveraged constraint-based stoichiometric modeling and kinetic ensemble modeling of the C. autoethanogenum metabolic network to provide a systematic in silico analysis of metabolic engineering interventions for 2,3-butanediol overproduction and low carbon substrate loss in dissipated CO2. Our analysis allowed us to identify and to assess comparatively the expected performances for a wide range of single, double, and triple interventions. Our analysis managed to individuate bottleneck reactions in relevant metabolic pathways when suggesting intervening strategies. Besides recapitulating intuitive and/or previously attempted genetic modifications, our analysis neatly outlined that interventions—at least partially—impinging on by-products branching from acetyl coenzyme A (acetyl-CoA) and pyruvate (acetate, ethanol, amino acids) offer valuable alternatives to the interventions focusing directly on the specific branch from pyruvate to 2,3-butanediol. IMPORTANCE Envisioning value chains inspired by environmental sustainability and circularity in economic models is essential to counteract the alterations in the global natural carbon cycle induced by humans. Recycling carbon-based waste gas streams into chemicals by devising gas fermentation bioprocesses mediated by acetogens of the genus Clostridium is one component of the solution. Carbon monoxide originates from multiple biogenic and abiogenic sources and bears a significant environmental impact. This study aims at identifying metabolic engineering interventions for increasing 2,3-butanediol production and avoiding carbon loss in CO2 dissipation via C. autoethanogenum fermenting a substrate comprising CO and H2. 2,3-Butanediol is a valuable biochemical by-product since, due to its versatility, can be transformed quite easily into chemical compounds such as butadiene, diacetyl, acetoin, and methyl ethyl ketone. These compounds are usable as building blocks to manufacture a vast range of industrially produced chemicals.

Enhancement of 2,3-Butanediol Production by Klebsiella pneumoniae: Emphasis on the Mediation of sRNA-SgrS on the Carbohydrate Utilization

Abstract: The demand for renewable energy is increasing. Klebsiella pneumoniae is one of the most promising strains to produce 2,3-butanediol (2,3-BD). Compared with chemical methods, the biological production of 2,3-BD has the characteristics of substrate safety, low cost, and low energy consumption. However, excessive glucose concentrations can cause damage to cells. Therefore, this study investigated the effect of sRNA-SgrS as a sugar transport regulator on the fermentative production of 2,3-BD by K. pneumoniae in response to sugar stress. We designed multiple mutants of K. pneumoniae HD79 to redistribute its carbon flux to produce 2,3-BD. It was found that the 2,3-BD yield of sgrS overexpressed strain decreased by 44% compared with the original strain. The results showed that a high concentration of sRNA-SgrS could accelerate the degradation of ptsG mRNA (encoding the glucose transporter EIICBGlc) and downregulate the expression levels of the budA gene (encoding the α-acetyllactate decarboxylase) and the budB gene (encoding the α-acetyllactate synthase) and budC gene (encoding the 2,3-BD dehydrogenase) but had no effect on the ack gene (encoding the acetate kinase) and the ldh gene (encoding the lactate dehydrogenase). It provides a theoretical basis and a technical reference for understanding the complex regulation mechanism of sRNA in microorganisms and the genetics and breeding in industrial fermentation engineering.

Metabolic Engineering of Corynebacterium Glutamicum for Efficient Production of Optically Pure (2R,3R)-2,3-Butanediol

Abstract: Abstract Background: 2,3-butanediol is an important platform compound which has a wide range of applications, involving in medicine, chemical industry, food and other fields. Especially the optically pure (2R,3R)-2,3-butanediol can be employed as an antifreeze agent and as the precursor for producing chiral compounds. However, some (2R,3R)-2,3-butanediol overproducing strains are pathogenic such as Enterobacter cloacae and Klebsiella oxytoca . Results: In this study, a (3R)-acetoin overproducing C. glutamicum strain, CGS9, was engineered to produce optically pure (2R,3R)-2,3-butanediol efficiently. Firstly, the gene bdhA from B. subtilis 168 was integrated into strain CGS9 and its expression level was further enhanced by using a strong promoter P sod and ribosome binding site (RBS) with high translation initiation rate, and the (2R,3R)-2,3-butanediol titer of the resulting strain was increased by 33.9%. Then the transhydrogenase gene udhA from E.coli was expressed to provide more NADH for 2,3-butanediol synthesis, which reduced the accumulation of the main byproduct acetoin by 57.2%. Next, a mutant atpG was integrated into strain CGK3, which increased the glucose consumption rate by 10.5% and the 2,3-butanediol productivity by 10.9% in shake-flask fermentation. Through fermentation engineering, the most promising strain CGK4 produced a titer of 144.9 g/L (2R,3R)-2,3-butanediol with a yield of 0.429 g/g glucose and a productivity of 1.10 g/L/h in fed-batch fermentation. The optical purity of the resulting (2R,3R)-2,3-butanediol surpassed 99%. Conclusion: To the best of our knowledge, this is the highest titer of optically pure (2R,3R)-2,3-butanediol achieved by GRAS strains, and the result has demonstrated that C. glutamicum is a competitive candidate for (2R,3R)-2,3-butanediol production.

Separating Mixture Glycols in DWC with Flow Rate-Composition Cascade Control Structure

Abstract: Dividing wall column (DWC) is an advanced technology which could decrease energy consumption. As the vertical wall inserted in DWC increases the degree of freedom of systems, the control becomes more difficult than the conventional columns. In this study, the mixture glycols (1, 2-propanediol+ 1, 3-butanediol+ 1, 4-butanediol) were taken as feed, an optimized DWC structure was proposed, and three control structures for DWC were tested by ± 10% feed disturbances. The temperature control structure (CS1) was hard to control the system while composition control structure (CS2) and flow rate-composition cascade control structure (CS3) had good performance on controlling the DWC. It showed that CS3 was superior to CS2, as CS3 could stabilize the DWC within 4 hours, and the maximum deviations of the three products, namely 1, 2-propanediol, 1, 3-butanediol and 1, 4-butanediol, were 0.07%, 1.44%, and 0.46% respectively, while CS2 could realize the stability within 7 hours, and the corresponding numbers were 0.35%, 2.78%, and 0.41%, respectively.

1,3‐Butanediol Mitigates High‐Salt Diet Induced Cognitive Impairment in Aging Mice

Abstract: Vascular dementia (VaD) is the second most common form of dementia after Alzheimer’s disease (AD). Hypertension is the second leading risk factor for developing neurodegenerative disease. A recent study reported that Dietary salt promotes cognitive impairment through tau phosphorylation. However, the effect of ketone bodies or the precursor molecule (1,3‐butanediol) supplementation on cognitive impairment in aged mice or the absence of high salt has not been investigated. The study tests the hypothesis that 1,3‐butanediol attenuates the progression of vascular dementia mediated through histone acetylation mechanism in hypertensive aged mice. To test this hypothesis, we employed old male and female wild‐type (WT) mice. For three months, the mice were fed with a high salt diet (HSD) (8%). HSD mice have supplemented with ketone bodies precursor, 1,3‐Butanediol (2%), and ketone bodies compound β‐hydroxybutyrate (BHB, 0.2%) in drinking water for 12 weeks. Blood pressure was measured after 12 weeks of treatment by tail‐cuff plethysmography. After 12 weeks, urine and tissues were collected. The results show that 1,3‐Butanediol or BHB treatment reduced blood pressure and miRNA‐128‐3p in aged hypertensive mice. Further, the data analysis revealed the loss of short‐term and long‐term memory in HSD‐hypertensive mice compared to HSD+BHB or HSD+ 1,3‐Butanediol mice. This mechanistic study shows that HSD+ 1,3‐Butanediol mice display an increased mitochondrial TFAM dependent upregulation of brain‐derived neurotrophic factor (BDNF) and nuclear receptor subfamily 4A (Nr4a) in the mouse hippocampus via activation of the H3K27ac acetylation at the promoter. In addition, the anti‐miR‐128 treatment restored neuroprotective function and reduced the dementia pathology in old‐hypertensive mice. The present data demonstrate that 1,3‐Butanediol or BHB increases ketone bodies production and improves cognitive functions in aged hypertensive mice and could be a novel nutritional intervention for vascular dementia treatment.