Fuel effects on PAH formation, toxicity and regulated pollutants: Detailed comparison of biodiesel blends with propanol, butanol and pentanol.

Blends of biodiesel and high-carbon alcohols have the potential to increase the rate of biofuel use in diesel engines, while reducing harmful and toxic compounds such as polycyclic aromatic hydrocarbons (PAHs). Since biodiesel and alcohols do not contain aromatic ingredients in their chemical structures, this study examined biodiesel blends with propanol, n-butanol, and 1-pentanol (5 %, 20 % and 35 % by vol.) and the effects of these aromatic-free fuels on regulated emissions, PAH formation and toxicity as compared to straight diesel fuel in a diesel engine operating at a constant speed and varying engine loads. PAH samples were meticulously processed and extensively analyzed using rigorous analytical chemistry methodology (gas chromatography-mass spectrometry (GC-MS)). Biodiesel and biodiesel-alcohol blends significantly reduced NOx emissions and the level of formation of PAHs and toxicity levels when compared to diesel fuel. Overall, adding 5 % alcohol to biodiesel decreased total PAH emissions. However, with the exception of 20 % propanol, adding 20 % and 35 % alcohol to biodiesel increased total PAH emissions as compared to neat biodiesel. In contrast, all blended fuels resulted in a decrease in the toxicity of PAH compounds (up to 70 %) and the percentage of higher-ring PAHs. Among higher alcohols, propanol blends stood out as reducing PAH formation as compared to n-butanol and pentanol blends. Overall, biodiesel-alcohol blends that emit less carcinogenic pollutants and primarily low-rings PAHs were found to be advantageous for reducing the likelihood of wetstacking in diesel engines under low load or cold operating conditions.

Qualitative and quantitative determination of butanol in latex paint by fast gas chromatography proton transfer reaction mass spectrometry.

Butanol is a common organic solvent used in latex paint, and one of its isomers, tert-butanol, is toxic and can cause potential harm to the human body. Therefore, it is of great significance to develop a qualitative and quantitative detection method for butanol isomers. In this study, we combined the advantages of rapid detection of proton transfer reaction mass spectrometry (PTR-MS) with the separation and qualitative capabilities of gas chromatography-mass spectrometry (GC-MS) to achieve the detection of isomers, building a fast gas chromatography proton transfer reaction mass spectrometry (FastGC-PTR-MS) equipment. Firstly, the developed technology was optimized using standard samples of several common volatile organic compounds. The retention times of acetonitrile, acetone, and alcohols were less than 50 s, and the retention times of the benzene series were less than 110 s, on the premise that these isomers could be basically separated (resolution R > 1.0). Compared with a commercial GC-MS equipment, the detection times were shortened by 5-6 times and 2-4 times, respectively. Then the FastGC-PTR-MS was applied to detect the isomers of butanol in latex paint. The results showed that the headspace of brand D latex paint mainly contained five substances: tert-butanol, n-butanol, acetaldehyde, methanol, and acetone. Tert-butanol and n-butanol could be completely separated (R > 1.5). The concentration of tert-butanol was 4.41 ppmv, far below the 100 ppmv maximum allowable workplace concentration. The developed FastGC-PTR-MS can be used for rapid qualitative and quantitative detection of butanol isomers in latex paint. The new equipment has the potential to play an important role in indoor environmental safety applications.

Use of hydrous ABE-glycerin-diesel microemulsions in a nonroad diesel engine - Performance and unignorable emissions.

Oversupply, extra energy consumption, and CO2 emissions from the refinery of biodiesel-derived glycerin (G) led to the consideration of its use as an alternative fuel. In this study, a nonroad diesel engine generator was employed to represent potential emissions under stringent regulated standards. G-diesel has been reported to reduce nitrogen oxides (NOx) and soot levels but increase CO and hydrocarbon emissions. A bio-producible acetone-butanol-ethanol (ABE) solution with multiple polarities was added to stabilize the glycerin and water in diesel examined in this study. A series of ABE-G-diesel blends were prepared to form the thermostable microemulsions. Four blends with small and well-dispersed bubbles were tested in the engine generator. The specific thermal efficiencies of the engine were slightly improved by using ABE-G from regular diesel due to better spray quality, longer ignition delay, and fuel-oxygen content that would enhance combustion. Meanwhile, the PM-NOx-CO emission trade-off in the previous study has been overcome by using ABE-G-diesel since the better fuel atomization and more premixed combustion were approached, as well as the lower and homogeneous in-cylinder temperature caused by water content and micro-explosion. However, the condensable particulate matter and nitro-PAHs were also observed and realized their unignorable contribution, which has not been regulated and even researched for the generators. Fortunately, the new fuels could inhibit both of them to a certain degree. Consequently, this study proposes using recyclable glycerin with a simple pretreatment mixed with ABE and diesel for greener nonroad diesel engine especially those equipped with low-grade aftertreatment.

Modeling microbial ethanol production by S. aureus, K. pneumoniae, and E. faecalis under aerobic/anaerobic conditions - applicability to laboratory cultures and real postmortem cases.

A quite intriguing subject being intensively researched in the forensic toxicology field is the source of postmortem determined blood ethanol concentration: antemortem ingestion or postmortem microbial production. Our previous research on microbial ethanol production has reported a quantitative relationship between the ethanol and the higher alcohols and 1-butanol produced by Escherichia coli, Clostridium perfrigens, and Clostridium sporogenes. In this contribution, we continue our research reporting on the following: (i) the patterns of ethanol, higher alcohols, and 1-butanol production by the microbes Klebsiella pneumoniae, Staphylococcus aureus, and Enterococcus faecalis (all being aerobic/facultative anaerobic species, common corpse's colonizers, and ethanol producers), under controlled laboratory conditions, (ii) the mathematical modeling, with simple mathematical equations, of the correlation between ethanol concentration and the other studied alcohols' concentrations, by performing multiple linear regression analysis of the results, and (iii) the applicability of the constructed models in microbial cultures developed under different temperature than that used to build the models, in denatured blood cultures and in real postmortem cases. The aforementioned alcohols were proved to be all indicators of ethanol production, both in qualitative and quantitative terms. 1-Propanol was the most significant alcohol in modeling microbial ethanol production, followed by methyl-butanol. The K. pneumoniae's models achieved the best scoring in applicability (E < 40%) compared to the S. aureus and E. faecalis models, both at laboratory microbial cultures at 37 °C and real postmortem cases. Overall, a noteworthy accuracy in estimating the microbial ethanol in cultures and autopsy blood is achieved by the employed simple linear models.

Quantification of Branched-Chain Alcohol-Based Biofuels and Other Fermentation Metabolites via High-Performance Liquid Chromatography.

As the consequences of climate change become apparent, metabolic engineers and synthetic biologists are exploring sustainable sources for transportation fuels. The design and engineering of microorganisms to produce bio-gasoline and other biofuels from renewable feedstocks can significantly reduce dependence on fossil fuels as well as lower the emissions of greenhouse gases. A significant amount of research over the past two decades has led to the development of microbial strains for the production of advanced fuel compounds. Crucial to these efforts are robust methods to quantify the amount of the biofuel compound being produced as well as the other metabolites that might be present during fermentation. Here, we provide a protocol for the quantification of branched-chain alcohols, specifically isobutanol and isopropanol, using high-performance liquid chromatography (HPLC).

Improving isobutanol tolerance and titers through EMS mutagenesis in Saccharomyces cerevisiae.

Improving yeast tolerance toward isobutanol is a critical issue enabling high-titer industrial production. Here, we used EMS mutagenesis to screen Saccharomyces cerevisiae with greater tolerance toward isobutanol. By this method, we obtained EMS39 with high-viability in medium containing 16 g/L isobutanol. Then, we metabolically engineered isobutanol synthesis in EMS39. About 2µ plasmids carrying PGK1p-ILV2, PGK1p-ILV3 and TDH3p-cox4-ARO10 were used to over-express ILV2, ILV3 and ARO10 genes, respectively, in EMS39 and wild type W303-1A. And the resulting strains were designated as EMS39-20 and W303-1A-20. Our results showed that EMS39-20 increased isobutanol titers by 49.9% compared to W303-1A-20. Whole genome resequencing analysis of EMS39 showed that more than 59 genes had mutations in their open reading frames or regulatory regions. These 59 genes are enriched mainly into cell growth, basal transcription factors, cell integrity signaling, translation initiation and elongation, ribosome assembly and function, oxidative stress response, etc. Additionally, transcriptomic analysis of EMS39-20 was carried out. Finally, reverse engineering tests showed that overexpression of CWP2 and SRP4039 could improve tolerance of S.cerevisiae toward isobutanol. In conclusion, EMS mutagenesis could be used to increase yeast tolerance toward isobutanol. Our study supplied new insights into mechanisms of tolerance toward isobutanol and enhancing isobutanol production in S. cerevisiae.

Enhanced isobutanol production by co-production of polyhydroxybutyrate and cofactor engineering.

Once cells have been used to produce biochemicals, there are only a few effective ways to utilize the residual cell mass, even though the utilization of leftover cells would aid in decreasing production costs. Here, a polyhydroxybutyrate (PHB) and isobutanol co-production system was designed to address this challenge. The addition of the PHB operon into Escherichia coli conferred a metabolic advantage for alcohol production, generating 1.14-fold more isobutanol. Furthermore, following nitrogen source optimization and cofactor engineering, the engineered E. coli strain produced 2-fold more isobutanol and 0.25 g/L PHB. Moreover, E. coli cells showed higher tolerance to isobutanol with the overexpression of PHB biosynthesis genes. This co-production system resulted in an increased biomass, higher glucose utilization, and lower acetate maintenance, leading to higher productivity regarding PHB and isobutanol yield. Thus, this novel system is applicable to future fermentation studies for the co-production of PHB and isobutanol.

Biobutanol production from sulfuric acid-pretreated red algal biomass by a newly isolated Clostridium sp. strain WK.

Marine biomass, especially the algal biomass, is currently considered to be one of the most potential candidates for biofuels conversion during the development of biomass utilization. In this study, a diluted sulfuric acid pretreatment method was established for biobutanol from red algal biomass Gelidium amansii using a newly isolated Clostridium sp. strain WK. Under the optimal condition of 2% sulfuric acid treated in 20 Min at 131 °C, the maximal hydrolysis percentage of biomass can reach up to 80.95%, and the biobutanol production was obtained to be 3.46 g/L with a yield of 0.20 g/g after the fermentation of biomass hydrolysate. This result demonstrated a 12.5-fold enhancement of conversion efficiency compared with the untreated control, which provides a new and efficient way to develop the biobutanol industry by utilizing the abundant, low cost, and carbohydrate-rich algal biomass.

Chemical typicality of South American red wines classified according to their volatile and phenolic compounds using multivariate analysis.

In this study, 83 wines representating four commercial categories: "Argentinean Malbec", "Brazilian Merlot", "Uruguayan Tannat" and "Chilean Carménère" were analyzed according to their phenolic and volatile compounds. The objective was to identify the chemical compounds that would typify each category. From approximately about 600 peaks obtained by chromatographic techniques, 169 were identified and 53 of them were selected for multivariate statistical analysis. Chilean Carménère was the best discriminated group by the methods applied in our study, followed by Argentinean Malbec. Brazilian Merlot mixed mainly with some Carménère, whileTannat mixed with all wines categories, especially Malbec. In general, Chilean Carménère wines can be characterized by a bluish color, higher amounts of sulphur dioxide, higher content of octanoic acid, isobutanol, ethyl isoamyl succinate and catechin and a smaller amount of quercetin. These data can contribute for further process of authenticity or typification of South American red wines.

Biobutanol production from sugarcane bagasse by Clostridium beijerinckii strains.

Acetone-butanol-ethanol (ABE) fermentation was performed with sugarcane bagasse (SCB) hydrolysate using Clostridium beijerinckii strains. A cost-effective SCB medium was developed with no enzymatic hydrolysis and no supplementation of extra carbon source or expensive nitrogen source. One of the C. beijerinckii strains studied was able to produce butanol with butanol productivity of 1.23 g/L/day with butanol yield of 0.18 g/g of sugars from the developed medium. High utilization rate of both glucose and xylose was observed in SCB medium during ABE fermentation. This study shows that SCB is a promising substrate for cellulosic biobutanol production.

A review on characteristics of food waste and their use in butanol production.

Biobutanol offers several advantages and a larger market, that make it a biofuel to be studied with great interest. In fact, butanol has an energy content similar to that of gasoline, and it can be used as an alternative fuel to gasoline. It is a biofuel that is safe for the environment. The optimization of the production of butanol thus appears as an attractive option. Butanol production from food waste (FW) is a process for carbon recovery and a method for solid waste recycling. Recently, the use of FW and food processing waste (FPW) as raw material for the production of butanol has attracted much interest. However, an efficient fermentation process is vital to improve the production of biobutanol. To the best of our knowledge, no review on butanol production from FW has been presented so far. Thus, this review focuses on the characteristics of FW and its potential to produce butanol. In addition, the main factors that affect their use for the production of butanol are also discussed.

Characterization of the oral breakdown, sensory properties, and volatile release during mastication of white bread.

The oral breakdown, sensory properties, and volatile release during mastication of white bread were investigated. The results of correlation analysis for white bread's physical properties and it's oral physiological parameters during chewing have elucidated that bread's physical properties determined the oral processing behavior. During chewing of white bread, 15 dominant ions with regularly changing patterns were monitored by proton transfer reaction-mass spectrometry (PTR-MS). These dominant ions derived from 32 volatile compounds were further confirmed by pure standards. Partial least squares regression (PLSR) analysis was used to explore the positive correlations between the sensory analysis and the dominant aroma compounds. Results have shown that 9 aroma compounds were predicted as the potent odorants contributing to the changes in aroma profiles. Finally, 3-hydroxy-2-butanone, 2-methyl-1-propanol, and heptanoic acid were confirmed as the key aroma compounds contributing to the changes in aroma profiles of white bread before and after chewing.

Production of butanol from biomass: recent advances and future prospects.

At present, diminishing oil resources and increasing environmental concerns have led to a shift toward the production of alternative biofuels. In the last few decades, butanol, as liquid biofuel, has received considerable research attention due to its advantages over ethanol. Several studies have focused on the production of butanol through the fermentation from raw renewable biomass, such as lignocellulosic materials. However, the low concentration and productivity of butanol production and the price of raw materials are limitations for butanol fermentation. Moreover, these limitations are the main causes of industrial decline in butanol production. This study reviews butanol fermentation, including the metabolism and characteristics of acetone-butanol-ethanol (ABE) producing clostridia. Furthermore, types of butanol production from biomass feedstock are detailed in this study. Specifically, this study introduces the recent progress on the efficient butanol production of "designed" and modified biomass. Additionally, the recent advances in the butanol fermentation process, such as multistage continuous fermentation, metabolic flow change of the electron carrier supplement, continuous fermentation with immobilization and recycling of cell, and the recent technical separation of the products from the fermentation broth, are described in this study.

Establishment of BmoR-based biosensor to screen isobutanol overproducer.

BACKGROUND: Isobutanol, a C4 branched-chain higher alcohol, is regarded as an attractive next-generation transport fuel. Metabolic engineering for efficient isobutanol production has been achieved in many studies. BmoR, an alcohol-regulated transcription factor, mediates a σ54-dependent promoter Pbmo of alkane monooxygenase in n-alkane metabolism of Thauera butanivorans and displays high sensitivity to C4-C6 linear alcohols and C3-C5 branched-chain alcohols. In this study, to achieve the high-level production of isobutanol, we established a screening system which relied on the combination of BmoR-based biosensor and isobutanol biosynthetic pathway and then employed it to screen isobutanol overproduction strains from an ARTP mutagenesis library. RESULTS: Firstly, we constructed and verified a GFP-based BmoR-Pbmo device responding to the isobutanol produced by the host. Then, this screening system was employed to select three mutants which exhibited higher GFP/OD600 values than that of wild type. Significantly, GFP/OD600 of mutant 10 was 190.7 ± 4.8, a 1.4-fold higher value than that of wild type. Correspondingly, the isobutanol titer of that strain was 1597.6 ± 129.6 mg/L, 2.0-fold higher than the wild type. With the overexpression of upstream pathway genes, the isobutanol production from mutant 10 reached 14.0 ± 1.0 g/L after medium optimization in shake flask. The isobutanol titer reached 56.5 ± 1.8 g/L in a fed-batch production experiment. CONCLUSIONS: This work screened out isobutanol overproduction strains from a mutagenesis library by using a screening system which depended on the combination of BmoR-based biosensor and isobutanol biosynthetic pathway. Optimizing fermentation condition and reinforcing upstream pathway could realize the increase of isobutanol production from the overproducer. Lastly, fed-batch fermentation of the mutant enhanced the isobutanol production to 56.5 ± 1.8 g/L.

Yellow top (Physaria fendleri) presscake: A novel substrate for butanol production and reduction in environmental pollution.

Yellow Top (Physaria fendleri) is a plant that belongs to the mustard family. This plant is used to produce seeds that are rich in hydroxy oil. After extraction of oil, the presscake is land filled. The seedcake is rich in polymeric sugars and can be used for various bioconversions. For the present case, the seedcake or presscake was hydrolyzed with dilute (0.50% [v/v]) H2 SO4 and enzymes to release sugars including glucose, xylose, galactose, arabinose, and mannose. Then, the hydrolyzate was used to produce acetone-butanol-ethanol (ABE). Using 100 gL-1 presscake (prior to pretreatment), 19.22 gL-1 of ABE was successfully produced of which butanol was the major product. In this process, an ABE productivity of 0.48 gL-1 h-1 was obtained. These results are superior to glucose fermentation to produce ABE in which an ABE productivity of 0.42 gL-1 h-1 was obtained. Use of Yellow Top to produce butanol has the following advantages: (i) it is an economic feedstock and is expected to produce butanol economically; (ii) it avoids pollution concerns when not land filled; and (iii) rate of ABE production is not inhibited when fermented this substrate. It is suggested that the potential of this feedstock be further explored by optimizing process parameters for this valuable fermentation. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2767, 2019.

The use of indigenous Saccharomyces cerevisiae and Starmerella bacillaris strains as a tool to create chemical complexity in local wines.

The performance of two vineyard strains, Saccharomyces cerevisiae SacPK7 and Starmerella bacillaris StbPK9, was evaluated in laboratory and pilot scale fermentations of Cretan grape must under the following inoculation schemes: single inoculation of SacPK7 (IS), simultaneous inoculation of StbPK9 and SacPK7 (SM), and sequential inoculation of StbPK9 followed by SacPK7 (SQ). Un-inoculated (spontaneous) fermentations (SP) and fermentations inoculated with control S. cerevisiae strains (CS) were also conducted as reference. Star. bacillaris not only did not restrict but also slightly promoted the growth of S. cerevisiae when the two strains were co-inoculated at equal quantities. On the contrary, the SQ inoculation scheme conferred a competitive advantage to Star. bacillaris over S. cerevisiae, which maximum population was reduced, while increased levels of Star. bacillaris were recorded. The fermentation kinetics were also affected, accordingly. The completion of fermentation was faster in SM, IS and CS ferments than in SQ and SP. Ethanol accumulation had a predominant role in the early death of Star. bacillaris, since its growth was similarly arrested irrespective of the dominating yeast species, the magnitude of yeast population or the availability of energy sources. Interestingly, the inoculation scheme applied significantly affected the chemical profiles of the resulting wines. SQ produced the most divergent chemical profile in sterile must, with glycerol, acetic acid, acetaldehyde, residual glucose, malic acid, ethyl acetate and higher alcohols being the key compounds affected by the prolonged activity of StbPK9. In pilot scale ferments, the indigenous S. cerevisiae produced twice as high levels of esters and higher alcohols compared to the commercial starter. Star. bacillaris further increased the levels of ethyl esters in the respective ferments. The use of a mixed S. cerevisiae/Star. bacillaris starter culture instead of S. cerevisiae alone enhanced the chemical complexity of Cretan local wine. The magnitude of differentiation was even higher when the addition of Star. bacillaris preceded that of S. cerevisiae. The highest divergence in analytical profiles was recorded between wines produced by native strain combinations and commercial S. cerevisiae. Present results show that the use of indigenous yeast formulations provides significant diversification to local wines, in line with the microbial terroir concept and recent observations that indigenous yeast strains may confer regional characters to wines.

Improvement of butanol production by the development and co-culture of C. acetobutylicum TSH1 and B. cereus TSH2.

Butanol fermentation comprises two successive and distinct stages, namely acidogenesis and solventogenesis. The current lack of clarity regarding the underlying metabolic regulation of fermentation impedes improvements in biobutanol production. Here, a proteomics study was performed in the acidogenesis phase, the lowest pH point (transition point), and the solventogenesis phase in the butanol-producing symbiotic system TSH06. Forty-two Clostridium acetobutylicum proteins demonstrated differential expression levels at different stages. The protein level of butanol dehydrogenase increased in the solventogenesis phase, which was in accordance with the trend of butanol concentration. Stress proteins were upregulated either at the transition point or in the solventogenesis phase. The cell division-related protein Maf was upregulated at the transition point. We disrupted the maf gene in C. acetobutylicum TSH1, and Bacillus cereus TSH2 was added to form a new symbiotic system. TSH06△maf produced 13.9 ± 1.0 g/L butanol, which was higher than that of TSH06 (12.3 ± 0.9 g/L). Butanol was furtherly improved in fermentation at variable temperature with neutral red addition for both TSH06 and TSH06△maf. The butanol titer of the maf deletion strain was higher than that of the wild type, although the exact mechanism remains to be determined.

Effect of ethanol and butanol on autotrophic growth of model homoacetogens.

Research efforts aimed at increasing ethanol and butanol productivity from syngas are currently gaining attention. For most model carboxydotrophic bacteria, production rates, yields and maximum product titres have been studied in detail, but little is known on alcohol toxicity in these bacteria. The aim of this work was to investigate the inhibitory effects of ethanol and butanol on the growth of Clostridium ljungdahlii PETC, C. carboxidivorans P7, and 'Butyribacterium methylotrophicum DSM3468'. Experiments to determine inhibitory effects due to product accumulation were carried out using a synthetic mixture of CO:CO2:H2 as a substrate. These conditions were chosen to mimic gaseous effluents of biomass and waste gasification plants. Inhibition effects were recorded as changes in growth parameters. No significant inhibition was observed for ethanol at concentrations below 15 g/L. The three species exhibited higher sensitivity to butanol. Half inhibition constants for butanol could be estimated for P7 (IC50 = 4.12 g/L), DSM3468 (IC50 = 1.79 g/L), and PETC (IC50 = 9.75 g/L). In conclusion, at least for the tested strains, alcohol toxicity is not an immediate handicap for increasing alcohol production of the tested homoacetogenic strains.

Transcriptional response to organic compounds from diverse gasoline and biogasoline fuel emissions in human lung cells.

Modern vehicles equipped with Gasoline Direct Injection (GDI) engine have emerged as an important source of particulate emissions potentially harmful to human health. We collected and characterized gasoline exhaust particles (GEPs) produced by neat gasoline fuel (E0) and its blends with 15% ethanol (E15), 25% n-butanol (n-But25) and 25% isobutanol (i-But25). To study the toxic effects of organic compounds extracted from GEPs, we analyzed gene expression profiles in human lung BEAS-2B cells. Despite the lowest GEP mass, n-But25 extract contained the highest concentration of polycyclic aromatic hydrocarbons (PAHs), while i-But25 extract the lowest. Gene expression analysis identified activation of the DNA damage response and other subsequent events (cell cycle arrest, modulation of extracellular matrix, cell adhesion, inhibition of cholesterol biosynthesis) following 4 h exposure to all GEP extracts. The i-But25 extract induced the most distinctive gene expression pattern particularly after 24 h exposure. Whereas E0, E15 and n-But25 extract treatments resulted in persistent stress signaling including DNA damage response, MAPK signaling, oxidative stress, metabolism of PAHs or pro-inflammatory response, i-But25 induced changes related to the metabolism of the cellular nutrients required for cell recovery. Our results indicate that i-But25 extract possessed the weakest genotoxic potency possibly due to the low PAH content.