Clostridium Acetobutylicum Research Articles

Artificial Cascade Transformation Biosystem for One-Pot Biomanufacturing of Odd-Numbered Neoagarooligosaccharides and d-Tagatose through Wiser Agarose Utilization.

The application of agarose oligosaccharides has garnered great attention, with their biological activities varying among different structures. However, it still meets a great bottleneck for the targeted production of odd-numbered neoagarooligosaccharides (NAOSs), such as neoagarotriose (NA3), due to the lack of one-step hydrolases. In this work, the α-agarase AgaA33 and β-galactosidase BgaD were synergistically used to prepare NA3 with agarose as a substrate. Additionally, an l-arabinose isomerase CaLAI from Clostridium acetobutylicum was characterized to valorize low-value byproducts (d-galactose) by forming d-tagatose, which exhibited good thermal stability without the need for additional metal ions. Under the optimal reaction conditions, the production of NA3 and d-galactose catalyzed by these three enzymes was 0.40 and 0.15 g/L, respectively. The artificial three-enzyme-based cascade transformation system not only achieved the highest production of NA3 until now but also allowed for the valorization of d-galactose, providing a wiser application route for agarose utilization.

Efficient biobutanol production via co-cultivation of Clostridium acetobutylicum and Bacillus cereus utilizing DES pretreated rice husk

Efficient biobutanol production via co-cultivation of Clostridium acetobutylicum and Bacillus cereus utilizing DES pretreated rice husk

Production of 1,4-butanediol through Clostridia C4 pathways

1,4-butanediol (1,4-BDO) is an important building block in the chemical industry that has been mainly produced from fossil fuels, but now biosynthesis of 1,4-BDO has received more and more attention due to environmental issues. The Clostridia C4 pathway produces an intermediate crotonyl-CoA which could be diverted to 1,4-BDO by 4-hydroxybutyryl-CoA dehydratase (4HBD). Here, we compared this pathway with other 1,4-BDO biosynthesis pathways and illustrated its potential advantages regarding cellular energy conservation and theoretical yield. Then, the feasibility of 1,4-BDO production in this way was tested by introducing a single 4HBD in Clostridium acetobutylicum that natively produced the C4 intermediate and a variety of aldehyde/alcohol dehydrogenases (AdhE). Five different 4HBD genes were screened and the Cbei-2100 gene from Clostridium beijerinckii was the most effective, producing 0.066 mg/mL of 1,4-BDO. To block the metabolic flux towards the main product butanol, disruption of butyryl-CoA dehydrogenase (Bcd) was tried but failed, while inactivation of its homologue (FAD/FMN-containing dehydrogenase, Fcd) obtained little effect. Alternatively, the electron-transferring flavoprotein EtfA coupled with Bcd was inactivated, and 1,4-BDO production was greatly increased to 0.182 mg/mL. In conclusion, this study demonstrated the feasibility of 1,4-BDO production through the Clostridia C4 pathway. Further blocking of the competing flux towards butanol would be effective to improve the production of in the future.

Butyrate as a growth factor of Clostridium acetobutylicum

The butyrate biosynthetic pathway not only contributes to electron management and energy generation in butyrate forming bacteria, but also confers evolutionary advantages to the host by inhibiting the growth of surrounding butyrate-sensitive microbes. While high butyrate levels induce toxic stress, effects of non-toxic levels on cell growth, health, metabolism, and sporulation remain unclear. Here, we show that butyrate stimulates cellular processes of Clostridium acetobutylicum, a model butyrate forming Firmicute. First, we deleted the 3-hydroxybutyryl-CoA dehydrogenase gene (hbd) from the C. acetobutylicum chromosome to eliminate the butyrate synthetic pathway and thus butyrate formation. A xylose inducible Cas9 cassette was chromosomally integrated and utilized for the one-step markerless gene deletions. Non-toxic butyrate levels significantly affected growth, health, and sporulation of C. acetobutylicum. Upon deleting spo0A, the gene of the master regulator of sporulation, Spo0A, and followed by butyrate addition experiments, we conclude that butyrate affects cellular metabolism through both Spo0A-dependent and independent mechanisms. We also deleted the hbd gene from the chromosome of the asporogenous C. acetobutylicum M5 strain lacking the pSOL1 plasmid to examine the potential involvement of pSOL1 genes on the observed butyrate effects. Addition of the precursor of butyrate biosynthesis crotonate to the hbd deficient M5 strain was used to probe the role of butyrate biosynthesis pathway in electron and metabolic fluxes. Finally, we found that butyrate addition can enhance the growth of the non-butyrate forming Clostridium saccharolyticum. Our data suggest that butyrate functions as a stimulator of cellular processes, like a growth factor, in C. acetobutylicum and potentially evolutionarily related Clostridium organisms.

Diverse YqeK Diadenosine Tetraphosphate Hydrolases Control Biofilm Formation in an Iron-Dependent Manner

YqeK is a bacterial HD-domain metalloprotein that hydrolyzes the putative second messenger diadenosine tetraphosphate (Ap4A). Elevated Ap4A levels are primarily observed upon exposure of bacteria to factors such as heat or oxidative stress and cause pleiotropic effects, including antibiotic sensitivity and disrupted biofilm formation. Ap4A thus plays a central role in bacterial physiology and metabolism, and its hydrolysis by YqeK is intimately linked to the ability of these microbes to cope with stress. Although YqeK is reported to hydrolyze Ap4A under aerobic conditions, all four existing crystal structures reveal an active site that consists of a diiron center, portraying a cryptic chemical nature for the active metallocofactor. This study examines two YqeK proteins from two ecologically diverse parent organisms: the obligate anaerobe Clostridium acetobutylicum and the facultative aerobe Bacillus halodurans. Both enzymes utilize Fe-based cofactors for catalysis, while under ambient or oxidative conditions, Bh YqeK hydrolyzes Ap4A more efficiently compared to Ca YqeK. This redox-dependent activity difference stems from the following two molecular mechanisms: the incorporation of mixed-metal, Fe-based bimetallic cofactors, in which the second metal is redox inert (i.e., Fe–Zn) and the upshift of the Fe–Fe cofactor reduction potentials. In addition, three strictly conserved, positively charged residues vicinal to the active site are critical for tuning Ap4A hydrolysis. In conclusion, YqeK is an Fe-dependent phosphohydrolase that appears to have evolved to permit Ap4A hydrolysis under different environmental niches (aerobic vs. anaerobic) by expanding its cofactor configuration and O2 tolerance.

Species-specific ribosomal RNA-FISH identifies interspecies cellular-material exchange, active-cell population dynamics and cellular localization of translation machinery in clostridial cultures and co-cultures.

Though dyes and fluorescent reporter proteins have played an essential role in identifying microbial species in co-cultures, we hypothesized that ribosomal RNA-fluorescence in situ hybridization (rRNA-FISH) could be adapted and applied to quantitatively probe complex interactions between organisms in synthetic consortia. Despite its maturity, several challenges existed before rRNA-FISH could be used to study Clostridium co-cultures of interest. First, species-specific probes for Clostridium acetobutylicum and Clostridium ljungdahlii had not been developed. Second, "state-of-the-art" labeling protocols were tedious and often resulted in sample loss. Third, it was unclear if FISH was compatible with existing fluorescent reporter proteins. We resolved these key challenges and applied the technique to co-cultures of C. acetobutylicum, C. ljungdahlii, and Clostridium kluyveri. We demonstrate that rRNA-FISH is capable of identifying rRNA/ribosome exchange between the three organisms and characterized rRNA localization patterns in each. In combination with flow cytometry, rRNA-FISH can capture sub-population dynamics in co-cultures.

Gut microbiota metabolic pathways: Key players in knee osteoarthritis development

ObjectiveTo confirm the causality of gut microbiota pathway abundance and knee osteoarthritis (KOA). MethodsMicrobial metabolic pathways were taken as exposures, with data from the Dutch Microbiome Project (DMP). Data on KOA from the UK Biobank were utilized as endpoints. In addition, we extracted significant and independent single nucleotide polymorphisms as instrumental variables. Two-sample Mendelian randomization (MR) analysis was applied to explore the causal relationship between gut microbiota pathway abundance and KOA, and MR-Egger and weighted median were used as additional validation of the MR results. Meanwhile, Cochran Q, MR-Egger intercept, MR-PRESSO, and leave-one-out were used to perform sensitivity analyses on the MR results. ResultsMR results showed that enterobactin biosynthesis, diacylglycerol biosynthesis I, Clostridium acetobutylicum acidogenic fermentation, glyoxylate bypass and tricarboxylic acid cycle were the risk factors for KOA. (OR = 1.13,95%CI = 1.04–1.23;OR = 1.12,95%CI = 1.04–1.20;OR = 1.14,95%CI = 1.04–1.26; OR = 1.06,95%CI = 1.00–1.12) However, adenosylcobalamin salvage from cobinamide I, hexitol fermentation to lactate formate ethanol and acetate, purine nucleotides degradation II aerobic, L tryptophan biosynthesis and inosine 5 phosphate biosynthesis III pathway showed significant protection against KOA. (OR = 0.93,95%CI = 0.86–1.00;OR = 0.94,95%CI = 0.88–1.00;OR = 0.91,95%CI = 0.86–0.97;OR = 0.95,95%CI = 0.92–0.99; OR = 0.91, 95%CI = 0.85–0.98) Further multiplicity and sensitivity analyses demonstrated the robustness of the results. ConclusionOur study identified specific metabolic pathways in gut microbiota that promote or inhibit KOA, which provides the most substantial evidence-based medical evidence for the pathogenesis and prevention of KOA.

Structural and biochemical insights into the molecular mechanism of N-acetylglucosamine/N-Acetylmuramic acid kinase MurK from Clostridium acetobutylicum

MurK is a MurNAc- and GlcNAc-specific amino sugar kinase, phosphorylates MurNAc and GlcNAc at the 6-hydroxyl group in an ATP-dependent manner, and contributes to the recovery of both amino sugars during the cell wall turnover in Clostridium acetobutylicum. Herein, we determined the crystal structures of MurK in complex with MurNAc, GlcNAc, and glucose, respectively. MurK represents the V-shaped fold, which is divided into a small N-terminal domain and a large C-terminal domain. The catalytic pocket is located within the deep cavity between the two domains of the MurK monomer. We mapped the significant enzyme-substrate interactions, identified key residues involved in the catalytic activity of MurK, and found that residues Asp77 and Arg78 from the β4-α2-loop confer structural flexibilities to specifically accommodate GlcNAc and MurNAc, respectively. Moreover, structural comparison revealed that MurK adopts closed-active conformation induced by the N-acetyl moiety from GlcNAc/MurNAc, rather than closed-inactive conformation induced by glucose, to carry out its catalytic reaction. Taken together, our study provides structural and functional insights into the molecular mechanism of MurK for the phosphorylation of both MurNAc and GlcNAc, sugar substrate specificity, and conformational changes upon sugar substrate binding.

AI-based screening of Clostridium acetobutylicum with high furfural tolerance and butanol production

Advances in strain breeding for butanol biosynthesis were quite limited because of physiological complexity of solventogenic Clostridia. Using AI, this study developed a high-throughput screening method for Clostridium acetobutylicum to find strains with inhibitor tolerance and high butanol production. A mutant library was generated from C. acetobutylicum ATCC 824 through ARTP mutagenesis and physiological traits were digitized using color indicators. The classification performance of Machine learning algorithms (PCA, PLS, SVM, ANN) were compared for different butanol-producing strains. Among 2000 strains screened, C. acetobutylicum Tust-f3 was identified, which could tolerate 4.5 g/L furfural and yield 10.5 g/L butanol from undetoxified lignocellulosic hydrolysate. Proteome analysis reveals that 38 proteins may play a crucial role. Subsequently, seven universal detoxification components for furfural were identified via heterologous expression in E. coli Genes CA_RS19590 and CA_RS08810 showed significant growth improvement (14.44 and 14.28-fold, respectively, compared to control). This study highlights the potential of machine learning in strain selection and breeding.

Microbial Induced Biotechnological Processes for Biofuel Production from Waste Organics Conversion

In the current era there are huge quantities of waste organic matter available, creating a big burden to the environment. To address these issues, researchers started to apply effective and microbial induced biotechnological processes that can mitigate these waste matters. In this context, different nature of microbial systems are involved in hydrolysing the waste organic material into fermentable sugar. These can be easily consumed by specific microbial systems like Saccharomyces cerevisiae MTCC 3821 and Clostridium acetobutylicum that produced bioethanol and biobutanol, respectively. Saccharomyces cerevisiae was cultured in specific media and incubated at rotary shaker with 150 rpm at 30°C for 72 to 96 hours. Ethanol concentrations from different waste matters were found in the range of 1.2-1.5 g.L-1. Ethanol synthesis was done by shake flask experiment with addition of glucose (50 g.L-1) to waste organic hydrolyzed solution. Non-glucose media produced less than 3 g.L-1 ethanol but glucose media produced 4.5 g.L-1. Next, Clostridium acetobutylicum was grown in culture media containing waste organics as sole carbon substrate with pH 7 and then was incubated in anaerobic conditions at 35°C for 72 hours, produced butanol (0.7 to 1.25 g.L-1). This research work promoted biofuels synthesis by keeping a waste mitigation strategy.

Sinusoidal control strategy applied to continuous stirred‐tank reactors: Asymptotic and exponential convergence

AbstractThe main goal of this proposal is to present a class of nonlinear controllers for regulation and set point changes in continuous chemical reactors. The proposed control law has in its mathematical structure a proportional term of the regulation error to provide closed‐loop stability and a sinusoidal term, which can compensate for the nonlinearities of the plant. The closed‐loop stability of the plant is demonstrated via Lyapunov analysis, which reveals an asymptotic convergence of the control output to the required set points. Furthermore, the analysis of the regulation error's dynamic under the considered assumptions leads us to conclude that exponential stability is also reached. The controller is implemented via numerical experiments in two examples to generalize the applicability of the proposed approach by considering continuous stirred‐tank reactors models. The first case considers autocatalytic chemical oscillatory reactions that induce chaotic behaviour. For the second case, a process of acetone, butanol, and ethanol (ABE) fermentation through Clostridium acetobutylicum is considered. The proposed strategy shows an adequate performance because it can reach the required set point without long time settings and overshoot. A comparison with a smooth sliding‐mode and a standard proportional‐integral (PI) controller indicates the advantages of the proposed control approach.

The fuel property and application prospect of water-containing biofuel from Clostridium acetobutylicum fermentation products

Biobutanol, a promising green alternative fuel, fermented from Clostridium acetobutylicum, while its high-cost and limited yield constraining its development. ABE (acetone-butanol-ethanol) and IBE (isopropanol-butanol-ethanol) are mixed fermentation products from non-edible biomass raw materials, using them together with water as alternative fuels will reduce industrial production costs and save fossil fuels. Therefore, this study conducted a multifaceted experimental evaluation on ABE/IBE mixed fuels with different water content, demonstrating that it has good water holding capacity when mixed with traditional fossil fuels. Taking ABE (3: 6: 1) as an example, its water holding capacity after mixing with diesel at 10–90 % is 0.37–7.83 % at 20 °C. Meanwhile, the particle size of ABE/IBE mixed fuels is about 2–30 nm, exhibiting a microemulsion with thermodynamic stability. The anhydrous or water-containing mixed fuel with the ratio of ABE (IBE) of 10%–50 % meets the range of the density and kinematic viscosity of diesel engine fuel. The mixed fuel is non-corrosive to copper without water, and a water content of about 3 % or higher will increase the risk of engine corrosion at 20 °C. Despite the addition of biofuel and water, studies on energy combustion performance and pollutant emission performance have found that appropriate addition of biofuel and water can produce higher power output and lower pollutant emissions than traditional fossil fuels, with ABE20W0.5 being the optimal. This study demonstrates the great potential of ABE and IBE as biofuels to achieve carbon neutrality goals, providing novel research direction for green alternative fuels in the future.

Enhanced biobutanol production with sustainable Co-substrates synergy from paper waste and garden waste with municipal biowaste

Synergy in the co-processing of lignocellulosic wastes and municipal biowaste (MB) can unlock their potential for biobutanol production. This study assessed the potential for biobutanol production through the co-processing of lignocellulosic waste and MB. Specifically, it compared the co-processing of paper waste with MB to that of garden waste and MB. Ethanol organosolv pretreatment served as a dual-function process for both pretreatment and detoxification purposes. Initial fermentation of hydrolysates from untreated paper waste using Clostridium acetobutylicum produced 0.9 g/L of acetone and ethanol but no detectable butanol. Organosolv pretreatment led to a significant increase in acetone and ethanol production but did not yield butanol. Co-processing paper waste with MB using organosolv pretreatment resulted in the production of 2.8–3.2 g/L butanol, along with increased acetone and ethanol production. Furthermore, co-processing a 1:1 (w/w) mixture of paper waste and MB under mild and severe pretreatment conditions produced 45.5 g and 43.4 g butanol, respectively, compared to 34.8 g and 14.4 g butanol when processing these waste streams separately. The study also explored the positive impact of co-processing garden waste with MB, a distinct lignocellulosic source, enhancing acetone-butanol-ethanol (ABE) yield by 27–40%. These findings highlight the potential of synergistic waste co-processing for achieving a more suitable balance of nutrients to enhance biobutanol and ABE production from biowastes. Additionally, the simultaneous treatment of lignocellulosic waste and municipal biowaste offers a simplified approach to waste processing, contributing to advancements in sustainable biomass utilization and bioenergy production.

Physicochemical and antibacterial properties of chitosan extracted from swimming crab shells and wooden grasshoppers using different extraction methods

Marine by-products and insects are among the sources of chitin used in chitosan production and increase the value of the product that may be used in the food industry. The conversion of chitin to chitosan requires proper extraction methods in order to minimise energy use and waste while also producing good-quality chitosan. This study aimed to evaluate different methods of extracting chitosan from two sources and to characterise its physicochemical and antibacterial properties. The study utilised two distinct chitosan sources, i.e. crab shells and wooden grasshoppers, as well as two distinct extraction methods, i.e. conventional and green chemistry methods. The yield, water-ash content, solubility, physicochemical properties as determined by infrared spectroscopy (FTIR), degree of deacetylation (DD), crystallinity (XRD), microstructure (SEM) and antibacterial activity were all evaluated for chitosan quality. The results indicated that the green chemistry extraction of crab shells (M2P1 treatment) produced the highest yield, solubility and crystallinity index of all treatments, with a DD of 60.9%. The functional groups and microstructure of chitosan were remarkably similar across all treatments. Antibacterial activity was determined using the microdilution method against Grampositive (Clostridium acetobutylicum) and Gram-negative (Escherichia coli) bacteria and the minimum inhibitory concentrations were identified, notably 2000 ppm for the green chemistry method. The green chemical extraction method using crab shells (M2P1) demonstrated that the extracted chitosan possessed beneficial physicochemical properties, especially on yield and solubility, and antimicrobial properties against both Gram-positive and Gram-negative bacteria. As such, based on the DD percentage and antibacterial activity, this implies that the extracted chitosan may be used as an alternative for the preservation of food in the food industry.

Chromosomal integration of the pSOL1 megaplasmid of Clostridium acetobutylicum for continuous and stable advanced biofuels production

Biofuel production by Clostridium acetobutylicum is compromised by strain degeneration due to loss of its pSOL1 megaplasmid. Here we used engineering biology to stably integrate pSOL1 into the chromosome together with a synthetic isopropanol pathway. In a membrane bioreactor continuously fed with glucose mineral medium, the final strain produced advanced biofuels, n-butanol and isopropanol, at high yield (0.31 g g−1), titre (15.4 g l−1) and productivity (15.5 g l−1 h−1) without degeneration.

Optimizing dilute acid pretreatment for enhanced recovery and co-fermentation of hexose and pentose sugars for ethanol and butanol production

A systematic study of dilute acid pretreatment of sugarcane bagasse was performed to optimize the recovery of hexose-rich solid and pentose-rich liquid while minimizing fermentation inhibitors. Pretreatment experiments evaluated the effects of pretreatment time (10–90 min) and acid concentration (0.5–3.0 % w/w). The optimized pretreatment (1.4 % w/w sulfuric acid, 56.4 min at 121 °C) led to a solid fraction with 60.4 % cellulose and a liquid stream rich in pentose, in which 92.7 % of sugars was recovered as xylose (17.8 g/L). Besides that, low concentrations of by-products (3 g/L – the sum of acetic and formic acid, furfural, and HMF) were identified in the liquid fraction. Both fractions were integrated into enzymatic processes for further co-fermentation of C5- and C6- sugars. Two fermentative routes were evaluated: simultaneous isomerization of xylose and co-fermentation of glucose and xylulose (SIcF) for 2G ethanol production and acetone-butanol-ethanol (ABE) fermentation of glucose and xylose for biobutanol, utilizing Saccharomyces cerevisiae and Clostridium acetobutylicum, respectively. SIcF yielded 0.48 gethanol/gsugars (24 g/L ethanol) with complete glucose consumption and 58.2 % xylose conversion. ABE fermentation produced 0.36 gABE/gsugars (15.7 g/L ABE and 9.5 g/L butanol), with 96 % glucose consumption and 29.6 % xylose conversion. The proof-of-concept demonstrated that the hydrolysates were satisfactorily fermentable by microorganisms, thus validating the pretreatment optimization for fermentation applications.

Gene essentiality in the solventogenic Clostridium acetobutylicum DSM 792.

Clostridium acetobutylicum is a leading candidate to synthesize valuable compounds like three and four carbons alcohols. Its ability to convert carbohydrates into a mixture of acetone, butanol, and ethanol as well as other chemicals of interest upon genetic engineering makes it an advantageous organism for the valorization of lignocellulose-derived sugar mixtures. Since, genetic optimization depends on the fundamental insights supplied by accurate gene function assignment, gene essentiality analysis is of great interest as it can shed light on the function of many genes whose functions are still to be confirmed. The data obtained in this study will be of great value for the research community aiming to develop C. acetobutylicum as a platform organism for the production of chemicals of interest.

Exopolysaccharides Synthesized by Lacticaseibacillus rhamnosus ŁOCK 0943: Structural Characteristics and Evaluation of Biological and Technological Properties

Lactic acid bacteria can synthesize extracellular exopolysaccharides (EPSs) that have versatile physicochemical and biological properties. In this paper, the EPSs synthesized by Lacticaseibacillus rhamnosus ŁOCK 0943 were characterized. Their structure, biological, and technological activity, as well as application potential, were analyzed. Chemical analysis showed that this strain produces mannan and β-1,6-glucan. Their emulsifying, antagonistic, and antioxidant properties, along with their prebiotic potential, were assessed. The analysis of the tested polymers’ ability to create a stable emulsion showed that their emulsifying activity depends mainly on the type of oily substance used. The analysis of the antagonistic activity revealed that these EPSs can inhibit the growth of yeasts (e.g., Candida albicans ATCC 10231) and potentially pathogenic bacteria (e.g., Clostridium acetobutylicum ŁOCK 0831, Enterococcus faecalis ATCC 29212). Moreover, EPSs positively influenced the growth of all tested probiotic bacteria. Furthermore, EPSs can be successfully used as a preservative in cosmetic products. The most effective results were obtained with the use of a 0.05% solution of a chemical preservative (bronopol) and 0.25 mg/mL of the EPSs.

Optimization of hydrogen and butanol production from agave guishe juice using Clostridium acetobutylicum ATCC824

Substrate selection is crucial to achieve cost feasibility in acetone-butanol-ethanol (ABE) fermentation. Therefore, the aim of this study was the optimization of hydrogen and butanol production by Clostridium acetobutylicum ATCC824 using Agave lechuguilla juice as low-cost substrate. Effects of C/N ratio, temperature and butyric acid addition were first evaluated conventionally (one factor at a time). Butyric acid addition showed an adverse effect in the process. In optimization studies, C/N ratio, temperature and guishe juice concentration were investigated using response surface methodology with a Box-Behnken design. Results revealed that temperature and juice concentration had significant influence, whether C/N ratio have no significant effect. Under optimum conditions, 5.96 g/L butanol and 4.37 L/L were produced. This study demonstrates the potential of guishe juice as a low-cost feedstock for hydrogen and butanol production, indicating a promising approach for further research into the full valorization of Agave lechuguilla through a biorefinery conceptualization.

Understanding the dewatering of fermentation-based 1,3-propanediol with acetone before cyclohexanone synthesis

Acetone can be produced by microbial fermentation using Clostridium acetobutylicum (acetone-n-butanol-ethanol fermentation, ABE fermentation), but it provides a mixture of acetone, 1-butanol, and ethanol. N-butanol and ethanol can be used as biofuels, but high-value applications of acetone generally require upgrading. Bio-based 1,3-propanediol is more readily produced by fermentation and separated at higher titers and has been used industrially. Therefore, the coupled upgrading of the two fermentation products can provide another avenue for high-value utilization of acetone. In this paper, salting-out extraction of 1,3-propanediol with acetone was carried out, and 1,3-propanediol and acetone would spontaneously form an organic phase, thus reducing the energy consumption in the 1,3-propanediol separation process. As the double alkylation reaction of acetone with equivalent 1,3-propanediol would generate cyclohexanone, the extraction of 1,3-propanediol with acetone can provide new insight into the process intensification (removing water, salt and water concentrated in the aqueous phase) for the 1,3-propanediol separation and conversion.