Biofiltration of n-butyl acetate with three packing material mixtures, with and without biochar.

Two cost-effective packing materials were used for n-butyl acetate removal in lab-scale biofilters, namely waste spruce root wood chips and biochar obtained as a byproduct from a wood gasifier. Three biofilters packed with spruce root wood chips: without biochar (SRWC), a similar one with 10% of biochar (SRWC-B) and that with 10% of biochar impregnated with a nitrogen fertilizer (SRWC-IB) showed similar yet differing maximum elimination capacities of 206 ± 27, 275 ± 21 and 294 ± 20 g m-3 h-1, respectively, enabling high pollutant removal efficiency (>95% at moderate loads) and stable performance. The original biochar adsorption capacity was high (208 ± 6 mgtoluene g-1), but near 70% of it was lost after a 300-day biofilter operation. By contrast, the exposed impregnated biochar drastically increased its adsorption capacity in 300 days (149 ± 7 vs. 17 ± 5 mgtoluene g-1). Colony forming unit (CFU) and microscopic analyses revealed significant packing material colonization by microorganisms and grazing fauna in all three biofilters with an acceptable pressure drop, up to 1020 Pa m-1, at the end of biofilter operation. Despite a higher price (14 vs. 123 €m-3), the application of the best performing SRWC-IB packing can reduce the total investment costs by 9% due to biofilter volume reduction.

Metabolically engineer Clostridium saccharoperbutylacetonicum for comprehensive conversion of acid whey into valuable biofuels and biochemicals.

As a byproduct of dairy production, the disposal of acid whey poses severe environmental challenges. Herein, an innovative solution involving metabolically engineering Clostridium saccharoperbutylacetonicum to convert all carbon sources in acid whey into sustainable biofuels and biochemicals was presented. By introducing several heterologous metabolic pathways relating to metabolisms of lactose, galactose, and lactate, the ultimately optimized strain, LM-09, exhibited exceptional performance by producing 15.1 g/L butanol with a yield of 0.33 g/g and a selectivity of 89.9%. Through further overexpression of alcohol acyl transferase, 2.7 g/L butyl acetate along with 6.4 g/L butanol was generated, resulting in a combined yield of 0.37 g/g. This study achieves the highest reported butanol titer and yield using acid whey as substrate in clostridia and marks pioneering production of esters using acid whey. The findings demonstrate an innovative bioprocess that enhances renewable feedstock biotransformation, thereby promoting economic viability and environmental sustainability of biomanufacturing.

Investigation of Physicochemical Characteristics of Aspergillus niger Biomass and Examination of Its Ability to Separate Butyl Acetate Isomers.

Aspergillus niger is a species of fungus that is widely found in natural ecosystems and has an important role in various industrial fields and is readily available. To study the adhesion of microbial cells to solid substrates and to improve their properties, physicochemical characterization of microorganisms is extremely important. For this purpose, in this study, the surface properties of A. niger biomass were determined at low cost and with high accuracy by inverse gas chromatography (IGC), a physicochemical characterization technique. IGC experiments were conducted between 303.2 and 328.2 K at infinite dilution. Among these temperatures, various organic solvent vapors were passed over the A. niger biomass considered as stationary phase and their retention behavior was studied. Using the raw data, net retention volumes were calculated and retention diagrams were drawn. From the linear retention diagrams, the dispersive surface energy was calculated according to Dorris-Gray (48.73-46.09 mJ/m2), Donnet-Park (47.12-44.50 mJ/m2), Schultz (46.88-42.45 mJ/m2), and Hamieh (76.42-64.06 mJ/m2) methods. With the IGC method, the acidity-basicity parameters of A. niger biomass were determined and it was found that the surface was basic ( K D / K A = 4.871 ). In the second part of this study, the butyl acetate isomer series, which are difficult to be separated by conventional methods, were effectively separated by the IGC method using A. niger stationary phase.

Simulation analysis of hazardous chemicals released from the furniture plant using ALOHA software.

The manufacturing of wooden furniture is extensive in Thailand's east. Hazardous chemicals were used in the wooden furniture industry's manufacturing process. Hazardous substances released into the surrounding atmosphere appear to have an impact on the environment and individuals. The ALOHA model is frequently used to assess hazardous chemicals released into the environment; this simulation model is an effective tool for modeling chemical compounds and detecting chemical disasters. It has a tremendous potential for preventing mishaps in potentially hazardous or emergency situations. Acetone and butyl acetate were extracted from the hardwood furniture business to identify accidents such as leaking, spillage, and evaporation. It is described as a highly poisonous, combustible, and explosive material. Toxic accident releases have negative implications for the surrounding areas. The goal of this work was to examine each accident using ALOHA software, and the computation of acetone and butyl acetate accidents was shown in this study. This project provides critical data for the furniture plant's chemical emergency rescue strategy as well as recommendations for emergency evacuation site decision-making.

A Gas Sensor with Enhanced Sensing Properties towards Butyl Acetate: Vascular Bundle Structure Zn2 SnO4 Derived from Maize Straw.

The development of butyl acetate sensors with high sensitivity and selectivity has been highly desirable for its harmful effects on human health. In this work, we developed a high-performance butyl acetate sensor based on vascular bundle structure Zn2 SnO4 nanomaterial derived from maize straw. The vascular bundle structure Zn2 SnO4 with higher specific surface area obtained by calcination to remove the maize straw template, plays the dual role of accelerating the diffusion of gas molecules and providing more active sites. Our research showed that the sensor had a response of 18 to 100 ppm butyl acetate at a working temperature of 250 °C, with a fast response recovery rate (18 s/25 s), which showed significant improvement compared to the Zn2 SnO4 sensor prepared without templates. The improved performance can be attributed to the cross-linked nanoparticle chains and gas collision mechanism of the sensor. These findings highlight the potential of our sensor for the detection of butyl acetate gas.

Electronic structure analysis of NiO quantum dot-modified jackfruit-shaped ZnO sensors and sensing properties investigation of their highly sensitive and selective for butyl acetate.

Detection of flammable, explosive and toxic butyl acetate helps to avoid accidents and protect health in industrial production. However, there are few reports on butyl acetate sensors, especially highly sensitive, low detection limit and highly selective ones. In this work, density functional theory (DFT) analyzes the electronic structure of sensing materials and the adsorption energy of butyl acetate. The effects of Ni element doping, oxygen vacancy constructions, and NiO quantum dot modifications on the modulation of the electronic structure of ZnO and on the adsorption energy of butyl acetate are investigated in detail. Based on the DFT analysis, the NiO quantum dot modified jackfruit-shaped ZnO is synthesized via thermal solvent method reduction. The NiO/ZnO sensor has a response 502.5 for 100 ppm butyl acetate with 100 ppb detection limit, and the response for 100 ppm butyl acetate is at least 6.2 times higher than 100 ppm methanol, 100 ppm benzene, 100 ppm triethylamine, 100 ppm isopropanol, 100 ppm ethyl acetate and 100 ppm formic acid. X-ray photoelectron spectroscopy (XPS) explores the change of oxygen vacancies in sensor accompanied with the addition of Ni element and reveales the reason for the change of oxygen vacancies.

Excess Enthalpies for Binary Mixtures of the Reactive System Acetic Acid + n-Butanol + n-Butyl Acetate + Water: Brief Data Review and Results at 313.15 K and Atmospheric Pressure.

The data on molar excess enthalpies, HmE, for the binary mixtures acetic acid + n-butanol, acetic acid + n-butyl acetate and n-butanol + n-butyl acetate at 313.15 K and atmospheric pressure were obtained with use of the C80 isothermal mixing calorimeter (Setaram). The correlation of the data was carried out using the NRTL model and Redlich-Kister equation. A comparative analysis with the literature data on all available binary subsystems of the quaternary system was carried out. Other thermodynamic properties (Cp,mE, SmE, ΔmixSm, GmE and ΔmixGm) of the binary systems were estimated using literature data and well-known formulas of classical thermodynamics.

Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO.

BACKGROUND: Ethyl acetate is a bulk chemical traditionally produced via energy intensive chemical esterification. Microbial production of this compound offers promise as a more sustainable alternative process. So far, efforts have focused on using sugar-based feedstocks for microbial ester production, but extension to one-carbon substrates, such as CO and CO2/H2, is desirable. Acetogens present a promising microbial platform for the production of ethyl esters from these one-carbon substrates. RESULTS: We engineered the acetogen C. autoethanogenum to produce ethyl acetate from CO by heterologous expression of an alcohol acetyltransferase (AAT), which catalyzes the formation of ethyl acetate from acetyl-CoA and ethanol. Two AATs, Eat1 from Kluyveromyces marxianus and Atf1 from Saccharomyces cerevisiae, were expressed in C. autoethanogenum. Strains expressing Atf1 produced up to 0.2 mM ethyl acetate. Ethyl acetate production was barely detectable (< 0.01 mM) for strains expressing Eat1. Supplementation of ethanol was investigated as potential boost for ethyl acetate production but resulted only in a 1.5-fold increase (0.3 mM ethyl acetate). Besides ethyl acetate, C. autoethanogenum expressing Atf1 could produce 4.5 mM of butyl acetate when 20 mM butanol was supplemented to the growth medium. CONCLUSIONS: This work offers for the first time a proof-of-principle that autotrophic short chain ester production from C1-carbon feedstocks is possible and offers leads on how this approach can be optimized in the future.

Surface analysis of sequential semi-solvent vapor impact (SAVI) for studying microstructural arrangements of poly(lactide-co-glycolide) microparticles.

Biodegradable poly(lactide-co-glycolide) (PLGA) microparticles have been used as long-acting injectable (LAI) drug delivery systems for more than three decades. Despite extensive use, few tools have been available to examine and compare the three-dimensional (3D) structures of microparticles prepared using different compositions and processing parameters, all collectively affecting drug release kinetics. Surface analysis after sequential semi-solvent impact (SASSI) was conducted by exposing PLGA microparticles to different semi-solvent in the liquid phase. The use of semi-solvent liquids presented practical experimental difficulties, particularly in observing the same microparticles before and after exposure to semi-solvents. The difficulties were overcome by using a new sequential semi-solvent vapor (SSV) method to examine the morphological changes of the same microparticles. The SASSI method based on SSV is called surface analysis of semi-solvent vapor impact (SAVI). Semi-solvents are the solvents that dissolve PLGA polymers depending on the polymer's lactide:glycolide (L:G) ratio. A sequence of semi-solvents was used to dissolve portions of PLGA microparticles in an L:G ratio-dependent manner, thus revealing different structures depending on how microparticles were prepared. Exposing PLGA microparticles to semi-solvents in the vapor phase demonstrated significant advantages over using semi-solvents in the liquid phase, such as in control of exposure conditions, access to imaging, decreasing the time for sequential exposure of semi-solvents, and using the same microparticles. The SSV approach for morphological analysis provides another tool to enhance our understanding of the microstructural arrangement of PLGA polymers. It will improve our comprehensive understanding of the factors controlling drug release from LAI formulations based on PLGA polymers.

Metabolic engineering of Escherichia coli for efficient biosynthesis of butyl acetate.

BACKGROUND: Butyl acetate is a versatile compound that is widely used in the chemical and food industry. The conventional butyl acetate synthesis via Fischer esterification of butanol and acetic acid using catalytic strong acids under high temperature is not environmentally benign. Alternative lipase-catalyzed ester formation requires a significant amount of organic solvent which also presents another environmental challenge. Therefore, a microbial cell factory capable of producing butyl acetate through fermentation of renewable resources would provide a greener approach to butyl acetate production. RESULT: Here, we developed a metabolically engineered strain of Escherichia coli that efficiently converts glucose to butyl acetate. A modified Clostridium CoA-dependent butanol production pathway was used to synthesize butanol which was then condensed with acetyl-CoA through an alcohol acetyltransferase. Optimization of alcohol acetyltransferase expression and redox balance with auto-inducible fermentative controlled gene expression led to an effective titer of 22.8 ± 1.8 g/L butyl acetate produced in a bench-top bioreactor. CONCLUSION: Building on the well-developed Clostridium CoA-dependent butanol biosynthetic pathway, expression of an alcohol acetyltransferase converts the butanol produced into butyl acetate. The results from this study provided a strain of E. coli capable of directly producing butyl acetate from renewable resources at ambient conditions.

Controlling selectivity of modular microbial biosynthesis of butyryl-CoA-derived designer esters.

Short-chain esters have broad utility as flavors, fragrances, solvents, and biofuels. Controlling selectivity of ester microbial biosynthesis has been an outstanding metabolic engineering problem. In this study, we enabled the de novo fermentative microbial biosynthesis of butyryl-CoA-derived designer esters (e.g., butyl acetate, ethyl butyrate, butyl butyrate) in Escherichia coli with controllable selectivity. Using the modular design principles, we generated the butyryl-CoA-derived ester pathways as exchangeable production modules compatible with an engineered chassis cell for anaerobic production of designer esters. We designed these modules derived from an acyl-CoA submodule (e.g., acetyl-CoA, butyryl-CoA), an alcohol submodule (e.g., ethanol, butanol), a cofactor regeneration submodule (e.g., NADH), and an alcohol acetyltransferase (AAT) submodule (e.g., ATF1, SAAT) for rapid module construction and optimization by manipulating replication (e.g., plasmid copy number), transcription (e.g., promoters), translation (e.g., codon optimization), pathway enzymes, and pathway induction conditions. To further enhance production of designer esters with high selectivity, we systematically screened various strategies of protein solubilization using protein fusion tags and chaperones to improve the soluble expression of multiple pathway enzymes. Finally, our engineered ester-producing strains could achieve 19-fold increase in butyl acetate production (0.64 g/L, 96% selectivity), 6-fold increase in ethyl butyrate production (0.41 g/L, 86% selectivity), and 13-fold increase in butyl butyrate production (0.45 g/L, 54% selectivity) as compared to the initial strains. Overall, this study presented a generalizable framework to engineer modular microbial platforms for anaerobic production of butyryl-CoA-derived designer esters from renewable feedstocks.

Differences in olfactory habituation between orthonasal and retronasal pathways.

The odorant arrives at nasal olfactory epithelium ortho- and retronasally. This experiment aimed to study the potential different olfactory habituation in orthonasal and retronasal pathways. 68 subjects were stimulated by constant airflow with an odor (50% phenethyl alcohol, PEA or 5% n-butyl acetate, BA) presented ortho- or retronasally. Participants rated the perceived odor intensity (0-10 points) per minute until the odor sensation disappeared. We also investigated the cross-habituation: when the subjects achieved full habituation, continue to rate odor intensity in a different pathway after instantly switching the odor stimulation pathway. The olfactory habituation curve was drawn. The differences of ratings between the orthonasal and retronasal olfaction at different time points and between male and female subjects were analyzed. The two odor intensity ratings decreased as the time extended, share the same "fast followed by slow" type. The ratings of orthonasal olfaction decreased faster than that of retronasal. The intensity rating of PEA of male retronasal approach was lower than that of female at the 5th min (p = 0.018). When orthonasal full habituation achieved, there was significant difference between the intensity ratings and the initial ratings of the retronasal stimulation pathway (p < 0.0001), and vice versa. We found obvious habituation as well as cross-habituation in both orthonasal and retronasal olfaction. The habituation of orthonasal olfaction was faster than that of retronasal olfaction. These different habituations were related to the gender.

Comprehensive evaluation for the one-pot biosynthesis of butyl acetate by using microbial mono- and co-cultures.

BACKGROUND: Butyl acetate has shown wide applications in food, cosmetics, medicine, and biofuel sectors. These short-chain fatty acid esters can be produced by either chemical or biological synthetic process with corresponding alcohols and acids. Currently, biosynthesis of short chain fatty acid esters, such as butyl butyrate, through microbial fermentation systems has been achieved; however, few studies regarding biosynthesis of butyl acetate were reported. RESULTS: In this study, three proof-of-principle strategies for the one-pot butyl acetate production from glucose through microbial fermentation were designed and evaluated. (1) 7.3 g/L of butyl acetate was synthesized by butanol-producing Clostridium acetobutylicum NJ4 with the supplementation of exogenous acetic acid; (2) With the addition of butanol, 5.76 g/L of butyl acetate can be synthesized by acetate-producing Actinobacillus succinogenes130z (ΔpflA); (3) Microbial co-culture of C. acetobutylicum NJ4 and A. succinogenes130z (ΔpflA) can directly produce 2.2 g/L of butyl acetate from glucose by using microbial co-culture system with the elimination of precursors. Through the further immobilization of A. succinogenes130z (ΔpflA), butyl acetate production was improved to 2.86 g/L. CONCLUSION: Different microbial mono- and co-culture systems for butyl acetate biosynthesis were successfully constructed. These strategies may be extended to the biosynthesis of a wide range of esters, especially to some longer chain ones.

Probing specificities of alcohol acyltransferases for designer ester biosynthesis with a high-throughput microbial screening platform.

Alcohol acyltransferases (AATs) enables microbial biosynthesis of a large space of esters by condensing an alcohol and an acyl-CoA. However, substrate promiscuity of AATs prevents microbial biosynthesis of designer esters with high selectivity. Here, we developed a high-throughput microbial screening platform that facilitates rapid identification of AATs for designer ester biosynthesis. First, we established a microplate-based culturing technique with in situ fermentation and extraction of esters. We validated its capability in rapid profiling of the alcohol substrate specificity of 20 chloramphenicol acetyltransferase variants derived from Staphylococcus aureus (CATSa ) for microbial biosynthesis of acetate esters with various exogeneous alcohol supply. By coupling the microplate-based culturing technique with a previously established colorimetric assay, we developed a high-throughput microbial screening platform for AATs. We demonstrated that this platform could not only probe the alcohol substrate specificity of both native and engineered AATs but also identify the beneficial mutations in engineered AATs for enhanced ester synthesis. We anticipate the high-throughput microbial screening platform provides a useful tool to identify novel wildtype and engineered AATs that have important roles in nature and industrial biocatalysis for designer bioester production.

Metabolic and Process Engineering of Clostridium beijerinckii for Butyl Acetate Production in One Step.

n-Butyl acetate is an important food additive commonly produced via concentrated sulfuric acid catalysis or immobilized lipase catalysis of butanol and acetic acid. Compared with chemical methods, an enzymatic approach is more environmentally friendly; however, it incurs a higher cost due to lipase production. In vivo biosynthesis via metabolic engineering offers an alternative to produce n-butyl acetate. This alternative combines substrate production (butanol and acetyl-coenzyme A (acetyl-CoA)), alcohol acyltransferase expression, and esterification reaction in one reactor. The alcohol acyltransferase gene ATF1 from Saccharomyces cerevisiae was introduced into Clostridium beijerinckii NCIMB 8052, enabling it to directly produce n-butyl acetate from glucose without lipase addition. Extractants were compared and adapted to realize glucose fermentation with in situ n-butyl acetate extraction. Finally, 5.57 g/L of butyl acetate was produced from 38.2 g/L of glucose within 48 h, which is 665-fold higher than that reported previously. This demonstrated the potential of such a metabolic approach to produce n-butyl acetate from biomass.

Developing Clostridium diolis as a biorefinery chassis by genetic manipulation.

Clostridium diolis can efficiently utilize various inexpensive, renewable resources such as crude glycerol and lignocellulosic biomass hydrolysate to produce bulk chemicals and fuels. However, its study has been impeded by the lack of efficient plasmids electro-transformation techniques. In this study, an efficient electroporation protocol for C. diolis was developed and two replicons functional in C. diolis were identified. After optimizing parameters, the electro-transformation efficiency was enhanced from 5 to 692 transformants/ug DNA. Moreover, metabolic engineering of C. diolis was performed as proof of concept for the first time. By simply overexpressing heterologous genes based on the replicable plasmids, the strain was engineered to improve productions of diol (1,3-propanediol) and n-alcohol (butanol), and to enable butyl acetate synthesis in vivo, respectively under different culture conditions. This work represented a milestone of breeding C. diolis using metabolic engineering, and paved the way for studying C. diolis on the molecular level.

Evolution of the key odorants and aroma profiles in traditional Laowuzeng baijiu during its one-year ageing.

Changes of key odorants and aroma profiles of Chinese Laowuzeng baijiu during its one-year ageing were determined by HS-SPME-AEDA and direct injection-AEDA (DI-AEDA). Ethyl hexanoate, (E,E)-2,4-decadienal, and 2-phenylethyl acetate showed the highest FD value (486) in all ageing stages. With regards to aroma profiles, fruity, floral, acidic, sweet/honey and cheesy aromas were enhanced during storage, while pickled vegetable, grain and alcoholic notes weakened during the ageing. Quantitation and OAVs showed that most of the aroma compounds (OAVs > 1), including ethyl esters, aldehydes, and acids, increased their contents within the same period, whereas nonanal, 2-phenylethyl acetate, ethyl benzoate, 4-ethylguaiacol, propanol and 3-methyl-1-butanol decreased in content after the storage of 365 days. Simulated aged samples, in which fresh samples were spiked with 18 compounds, were examined by triangle tests, which indicated that the "fruity" compounds were crucial for maintaining the special aroma profile of an aged sample.

Combining untargeted, targeted and sensory data to investigate the impact of storage on food volatiles: A case study on strawberry juice.

An integrated science-based approach, combining analytical and sensorial data and different data analysis methods, proved successful to study the impact of storage time, storage temperature and oxygen availability on strawberry juice volatiles and allowed to get a multi-perspective view on these changes. An untargeted GC-MS approach showed that the volatile fraction of shelf-stable strawberry juice clearly changed during ambient storage and that oxygen availability (linked to the type of bottle) had a limited effect. To gain further insight, several characteristic aroma compounds were quantified during storage at ambient (20 °C) and accelerated (28-42 °C) temperatures, kinetic parameters were estimated and odour activity values were calculated. The kinetic parameters showed that all characteristic aroma compounds changed significantly during storage at all temperatures and that the rate of change in some compounds was accelerated by storage at higher temperatures. The observed changes in strawberry juice volatiles caused sensorial differences between non-stored and 20 °C stored samples as shown by the sensory evaluations and odour activity values.

Utilization of deep-sea microbial esterase PHE21 to generate chiral sec-butyl acetate through kinetic resolutions.

We previously identified and characterized 1 novel deep-sea microbial esterase PHE21 and used PHE21 as a green biocatalyst to generate chiral ethyl (S)-3-hydroxybutyrate, 1 key chiral chemical, with high enantiomeric excess and yield through kinetic resolution. Herein, we further explored the potential of esterase PHE21 in the enantioselective preparation of secondary butanol, which was hard to be resolved by lipases/esterases. Despite the fact that chiral secondary butanols and their ester derivatives were hard to prepare, esterase PHE21 was used as a green biocatalyst in the generation of (S)-sec-butyl acetate through hydrolytic reactions and the enantiomeric excess, and the conversion of (S)-sec-butyl acetate reached 98% and 52%, respectively, after process optimization. Esterase PHE21 was also used to generate (R)-sec-butyl acetate through asymmetric transesterification reactions, and the enantiomeric excess and conversion of (R)-sec-butyl acetate reached 64% and 43%, respectively, after process optimization. Deep-sea microbial esterase PHE21 was characterized to be a useful biocatalyst in the kinetic resolution of secondary butanol and other valuable chiral secondary alcohols.

Kinetics of Low Temperature Polyester Dyeing with High Molecular Weight Disperse Dyes by Solvent Microemulsion and AgroSourced Auxiliaries.

This work focused on the evaluation of the kinetics of dyeing polyester fabrics with high molecular weight disperse dyes, at low temperature by solvent microemulsion. This study also compared the effect of two non-toxic agro-sourced auxiliaries (o-vanillin and coumarin) using a non-toxic organic solvent. A dyeing bath consisting of a micro-emulsion system involving a small proportion of n-butyl acetate was used, and the kinetics of dyeing were analysed at four temperatures (83, 90, 95 and 100 °C). Moreover, the dyeing rate constants, correlation coefficient and activation energies were proposed for this system. It was found that o-vanillin yielded higher dye absorption levels than coumarin, leading to exhaustions of 88% and 87% for Disperse Red 167 and Disperse Blue 79, respectively. K/S values of dyed polyester were also found to be higher for dye baths containing o-vanillin with respect to the ones with coumarin. In terms of hot pressing fastness and wash fastness, generally no adverse influence on fastness properties was reported, while o-vanillin showed slightly better results compared to coumarin.