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Pyrolysis, a thermal decomposition without oxygen, is a promising technology for transportable liquids from whole fractions of lignocellulosic biomass. However, due to the hydrophilic products of pyrolysis, the liquid oils have undesirable physicochemical characteristics, thus requiring an additional upgrading process. Biological upgrading methods could address the drawbacks of pyrolysis by utilizing various hydrophilic compounds as carbon sources under mild conditions with low carbon footprints. Versatile chemicals, such as lipids, ethanol, and organic acids, could be produced through microbial assimilation of anhydrous sugars, organic acids, aldehydes, and phenolics in the hydrophilic fractions. The presence of various toxic compounds and the complex composition of the aqueous phase are the main challenges. In this review, the potential of bioconversion routes for upgrading the aqueous phase of pyrolysis oil is investigated with critical challenges and perspectives. Graphical Abstract

Violacein, a blue-violet compound with a wide range of beneficial bioactivities, is an attractive product for microbial production. Currently, violacein production has been demonstrated in several sugar heterotrophs through metabolic engineering; however, the cost of production remains an obstacle for business ventures. To address this issue, the development of host strains that can utilize inexpensive alternative substrates to reduce production costs would enable the commercialization of violacein. In this study, we engineered a facultative methylotroph, Methylorubrum extorquens AM1, to develop a methanol-based platform for violacein production. By optimizing expression vectors as well as inducer concentrations, 11.7 mg/L violacein production was first demonstrated using methanol as the sole substrate. Considering that unidentified bottlenecks for violacein biosynthesis in the shikimate pathway of M. extorquens AM1 would be difficult to address using generic metabolic engineering approaches, random mutagenesis and site-directed mutagenesis were implemented, and a 2-fold improvement in violacein production was achieved. Finally, by co-utilization of methanol and acetate, a remarkable enhancement of violacein production to 118 mg/L was achieved. Our results establish a platform strain for violacein production from non-sugar feedstocks, which may contribute to the development of an economically efficient large-scale fermentation system for violacein production.

Sustainable production of chemicals and materials from renewable non-food biomass using biorefineries has become increasingly important in an effort toward the vision of 'net zero carbon' that has recently been pledged by countries around the world. Systems metabolic engineering has allowed the efficient development of microbial strains overproducing an increasing number of chemicals and materials, some of which have been translated to industrial-scale production. Fermentation is one of the key processes determining the overall economics of bioprocesses, but has recently been attracting less research attention. In this Review, we revisit and discuss factors affecting the competitiveness of bacterial fermentation in connection to strain development by systems metabolic engineering. Future perspectives for developing efficient fermentation processes are also discussed.

Biodegradable polyhydroxybutyrate (PHB) can be produced from methane by some type II methanotroph such as the genus Methylocystis. This study presents the comparative genomic analysis of a newly isolated methanotroph, Methylocystis sp. MJC1 as a biodegradable PHB-producing platform strain. Methylocystis sp. MJC1 accumulates up to 44.5% of PHB based on dry cell weight under nitrogen-limiting conditions. To facilitate its development as a PHB-producing platform strain, the complete genome sequence of Methylocystis sp. MJC1 was assembled, functionally annotated, and compared with genomes of other Methylocystis species. Phylogenetic analysis has shown that Methylocystis parvus to be the closest species to Methylocystis sp. MJC1. Genome functional annotation revealed that Methylocystis sp. MJC1 contains all major type II methanotroph biochemical pathways such as the serine cycle, EMC pathway, and Krebs cycle. Interestingly, Methylocystis sp. MJC1 has both particulate and soluble methane monooxygenases, which are not commonly found among Methylocystis species. In addition, this species also possesses most of the RuMP pathway reactions, a characteristic of type I methanotrophs, and all PHB biosynthetic genes. These comparative analysis would open the possibility of future practical applications such as the development of organism-specific genome-scale models and application of metabolic engineering strategies to Methylocystis sp. MJC1.

By facilitating electron transfer to the hydroxylase diiron center, MMOR-a reductase-serves as an essential component of the catalytic cycle of soluble methane monooxygenase. Here, the X-ray structure analysis of the FAD-binding domain of MMOR identified crucial residues and its influence on the catalytic cycle.

In this study, we performed a kinetic analysis of CO-dependent H2 production by metabolically engineered Thermococcus onnurineus NA1, MC01 in terms of the cell activity as well as mass transfer rate. We conducted continuous cultures with varying dilution rate, CO flow rate, or agitation speed. Despite oscillations in cell density, the cultures reached steady states at all operating conditions. As the dilution rate increased from 0.1 to 0.3 h−1, specific activity of H2 production (SAHP) and volumetric cell production rate were linearly increased. Also, the SAHP remained almost constant at the fixed dilution rate of 0.3 h−1 even though the CO transfer rate was changed by adjusting the CO flow rate or the agitation speed. This relationship could be expressed as a typical Luedeking-Piret model, implying that high cell density culture with a sufficient growth rate is essential to obtain higher H2 productivity. On the other hand, more elevated CO transfer at the same dilution rate improved H2 production rate (HPR) by the increase of the cell density, not in the rise of SAHP. Through the continuous culture, 108 mmol/g-cell/h and 121 mmol/L/h of SAHP and HPR, respectively, could be achieved at a dilution rate of 0.3 h−1 with CO supply rate of 0.07 vvm and agitation speed of 700 rpm. Considering high H2 production activity and long-term stability of the strain over 1,000 h, MC01 is confirmed to have an outstanding potential for CO-dependent H2 production.

Fueled by the recognition of hydrogen as a promising renewable energy source for the future, there have been many attempts to find greener and more economical ways for its production from various sources. In this study, Methylomonas sp. DH-1, a type I methanotroph, was found to produce hydrogen using methane as a sole carbon source, under micro-aerobic conditions; this is analogous to the partial oxidation of methane in a thermochemical process based on metal catalysts. Flask cultures of Methylomonas sp. DH-1 were used to investigate the effects of different culture conditions on hydrogen production, including oxygen levels, methane/oxygen ratios, and initial cell densities. Methylomonas sp. DH-1 could produce hydrogen at an oxygen level below 4%, regardless of the methane content in the flask, implying that the critical factor for hydrogen production is the oxygen level, rather than the methane/oxygen ratio. Moreover, Methylomonas sp. DH-1 shows reversibility in hydrogen production and uptake, because the strain produces hydrogen under micro-aerobic conditions, uptakes it when the oxygen levels increase, and restores the hydrogen production capability when conditions become microaerobic again. Under initial conditions of 30% methane, 70% air, and an OD600nm of 6, hydrogen production was 26.87 μmol and its yields per methane and dry cell weight were 14.98 mmol-H2/mol-CH4 and 101.53 μmol-H2/g DCW, respectively, after 24 h of cultivation.

By facilitating electron transfer to the hydroxylase diiron center, MMOR-a reductase-serves as an essential component of the catalytic cycle of soluble methane monooxygenase. Here, the X-ray structure analysis of the FAD-binding domain of MMOR identified crucial residues and its influence on the catalytic cycle.

Since the 20th century, plastics that are widely being used in general life and industries are causing enormous plastic waste problems since improperly discarded plastics barely degrade and decompose. Thus, the demand for polyhydroxyalkanoates (PHAs), biodegradable polymers with material properties similar to conventional petroleum-based plastics, has been increased so far. The microbial production of PHAs is an environment-friendly solution for the current plastic crisis, however, the carbon sources for the microbial PHA production is a crucial factor to be considered in terms of carbon-neutrality. One‑carbon (C1) resources, such as methane, carbon monoxide, and carbon dioxide, are greenhouse gases and are abundantly found in nature and industry. C1 resources as the carbon sources for PHA production have a completely closed carbon loop with much advances; i) fast carbon circulation with direct bioconversion process and ii) simple fermentation procedure without sterilization as non-preferable nutrients. This review discusses the biosynthesis of PHAs based on C1 resource utilization by wild-type and metabolically engineered microbial host strains via biorefinery processes.

Electrofermentation actively regulates the bacterial redox state, which is essential for bioconversion and has been highlighted as an effective method for further improvements of the productivity of either reduced or oxidized platform chemicals. 1,3-Propanediol (1,3-PDO) is an industrial value-added chemical that can be produced from glycerol fermentation. The bioconversion of 1,3-PDO from glycerol requires additional reducing energy under anoxic conditions. The cathode-based conversion of glycerol to 1,3-PDO with various electron shuttles (2-hydroxy-1,4-naphthoquinone, neutral red, and hydroquinone) using Klebsiella pneumoniae L17 was investigated. The externally poised potential of -0.9 V vs. Ag/AgCl to the cathode increased 1,3-PDO (35.5±3.1 mm) production if 100 μm neutral red was used compared with non-bioelectrochemical system fermentation (23.7±2.4 mm). Stoichiometric metabolic flux and transcriptional analysis indicated a shift in the carbon flux toward the glycerol reductive pathway. The homologous overexpression of glycerol dehydratase (DhaB) and 1,3-PDO oxidoreductase (DhaT) enzymes synergistically enhanced 1,3-PDO conversion (39.3±0.8 mm) under cathode-driven fermentation. Interestingly, a small current uptake (0.23 mmol of electrons) caused significant metabolic flux changes with a concomitant increase in 1,3-PDO production. This suggests that both an increase in 1,3-PDO production and regulation of the cellular metabolic pathway are feasible by electrode-driven control in cathodic electrofermentation.

Native 3-hydroxybutyrate dehydrogenase from Alcaligenes faecalis can catalyze the reversible reduction of acetoacetate, a four carbon chain oxo acid. This enzyme has been engineered to enable the reduction of levulinic acid, with one carbon longer than acetoacetate. In this study, the native and engineered enzymes were subjected to the catalysis of oxo acids with a carbon chain length of 3 to 8, in order to examine the capability of the enzyme to work on various platform chemicals. The engineered enzyme could reduce the C7 and C8 oxo acids whereas the wild-type had no activity on these substrates. Docking simulation has indicated Tyr155 and Ser142 are key residues for the catalysis. In addition, stable hydrogen bond formation between Gln196 and the substrates affects the turnover rate. Mutation sites in the engineered enzyme were focused on creating larger active site volume for substrates with extended chain lengths. Both qualitative and quantitative structural basis for the enzyme substrate specificity on alpha, beta, gamma and omega hydroxy acids could be elucidated.

We investigated the algorithm analysis of the bubble generation in a microfluidic device to study the effect of the surface tension and the flow rate on the microbubble size. For the analysis of the surface tension, five different solutions were used: 3.5% brine, mineral oil, 1% polyethylene glycol (PEG) 400, 1% tween 80, and 1% triton X-100. The various flow rates were also employed: 5~15 µL/min for the liquid and 100~200 mL/min for the gas phase. The size of the bubble was measured via the algorithm analysis and the bubble defect was also detected by c chart. We observed that the microbubble size was affected by the flow rates of solution and the gas. Hence, we developed an equation to estimate the size through the flow rate ratio between the solution and gas phase, showing that the microbubble size could be controlled by the liquid properties or the flow rates. Therefore, this algorithm-based microfluidic device could be a powerful tool for generating gas micro-bubbles in a controlled manner.

Propane is the main component of liquefied petroleum gas and is derived from crude oil processing. Methanotrophic bacteria can convert various alkanes using methane monooxygenase enzyme to primary alcohols. These are further oxidized to various aldehydes by alcohol dehydrogenases or methanol dehydrogenases. In this study, 2-propanol was produced from propane using the whole cells of Methylosinus trichosporium OB3b, Methylomicrobium alcaliphilum 20Z, and Methylomonas sp. DH-1 as the biocatalysts. The biocatalytic process of converting propane to 2-propanol was optimized by the use of several inhibitors and additives, such as EDTA, sodium phosphate, and sodium formate to prevent oxidation of 2-propanol to acetone and to enhance conversion of propane to propanol. The maximum titer of 2-propanol was 0.424 g/L, 0.311 g/L, and 0.610 g/L for Methylomonas sp. DH-1, M. alcaliphilum 20Z, and M. trichosporium OB3b whole cells, respectively. These results showed that type I and type II methanotrophs could be used as the potent biocatalyst for conversion of propane to propanol.

A solvent-free co-production process for biodiesel and glycerol carbonate (GC) was developed based on enzyme (Novozyme 435). Glycerol obtained from transesterification reactions could be directly converted to GC. The reactions were optimized using the response surface methodology (RSM) separately. The optimal factors, which are enzyme loading, molar ratio of dimethyl carbonate to soybean oil, water content, and reaction temperature, were derived from the response optimizer. The results of optimization for biodiesel and GC production were re-analyzed by overlapping the contour plots of the factors. The following were the optima obtained: enzyme loading of 25 g/L, molar ratio of 15:1, water content of 0.082% (v/v), and reaction temperature of 59.65 °C. When the co-production of biodiesel and GC was conducted using the optima, biodiesel and GC production were achieved with conversions of 97.6% and 95.5%, respectively.

The production of industrial chemicals from renewable biomass resources is a promising solution to overcome the society’s dependence on petroleum and to mitigate the pollution resulting from petroleum processing. Klebsiella pneumoniae is a nutritionally versatile bacterium with numerous native pathways for the production of well-known and industrially important platform chemicals derived from various sugars. Genomic sequence analyses have shown that the K. pneumoniae genome has a high similarity with that of Escherichia coli, the most studied organism, which is used in industrial biotechnology processes for fuel and chemical production. Hence, K. pneumoniae can be considered as a promising platform microorganism that can be metabolically engineered for the high-level production of bio-based chemicals. This review highlights the substrate metabolism and the metabolic engineering strategies developed in K. pneumoniae for the production of bio-based chemicals.

Background Ectoine (1,3,4,5-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid) is an attractive compatible solute because of its wide industrial applications. Previous studies on the microbial production of ectoine have focused on sugar fermentation. Alternatively, methane can be used as an inexpensive and abundant resource for ectoine production by using the halophilic methanotroph, Methylomicrobium alcaliphilum 20Z. However, there are some limitations, including the low production of ectoine from methane and the limited tools for the genetic manipulation of methanotrophs to facilitate their use as industrial strains. Results We constructed M. alcaliphilum 20ZDP with a high conjugation efficiency and stability of the episomal plasmid by the removal of its native plasmid. To improve the ectoine production in M. alcaliphilum 20Z from methane, the ectD (encoding ectoine hydroxylase) and ectR (transcription repressor of the ectABC-ask operon) were deleted to reduce the formation of by-products (such as hydroxyectoine) and induce ectoine production. When the double mutant was batch cultured with methane, ectoine production was enhanced 1.6-fold compared to that obtained with M. alcaliphilum 20ZDP (45.58 mg/L vs. 27.26 mg/L) without growth inhibition. Notably, a maximum titer of 142.32 mg/L was reached by the use of an optimized medium for ectoine production containing 6% NaCl and 0.05 μM of tungsten without hydroxyectoine production. This result demonstrates the highest ectoine production from methane to date. Conclusions Ectoine production was significantly enhanced by the disruption of the ectD and ectR genes in M. alcaliphilum 20Z under optimized conditions favoring ectoine accumulation. We demonstrated effective genetic engineering in a methanotrophic bacterium, with enhanced production of ectoine from methane as the sole carbon source. This study suggests a potentially transformational path to commercial sugar-based ectoine production. Graphical Abstract

Cupriavidus necator, a versatile microorganism found in both soil and water, can have both heterotrophic and lithoautotrophic metabolisms depending on environmental conditions. C. necator has been extensively examined for producing Polyhydroxyalkanoates (PHAs), the promising polyester alternatives to petroleum-based synthetic polymers because it has a superior ability for accumulating a considerable amount of PHAs from renewable resources. The development of metabolically engineered C. necator strains has led to their application for synthesizing biopolymers, biofuels and biochemicals such as ethanol, isobutanol and higher alcohols. Bio-based processes of recombinant C. necator have made much progress in production of these high-value products from biomass wastes, plastic wastes and even waste gases. In this review, we discuss the potential of C. necator as promising platform host strains that provide a great opportunity for developing a waste-based circular bioeconomy.

Auto-generative high pressure digestion (AHPD) and hydrogen-injecting digestion (HID) have been introduced to directly produce high CH4-content biogas from anaerobic digester. However, each approach has its own technical difficulties (pH changes), and practical issues (high cost of H2) to obtain > 90% CH4 containing biogas, particularly, from the high-strength waste like food waste (FW). To overcome this problem, in this study, AHPD and HID were integrated, which can offset each drawback but maximize its benefit. Substrate concentration of FW tested here was 200 g COD/L, the highest ever applied in AHPD and HID studies. At first, the reactor was operated by elevating the autogenerative pressure from 1 to 3, 5, and 7 bar without H2 injection. With the pressure increase, the CH4 content in the biogas gradually increased from 52.4% at 1 bar to 77.4% at 7 bar. However, a drop of CH4 production yield (MPY) was observed at 7 bar, due to the pH drop down to 6.7 by excess CO2 dissolution. At further operation, H2 injection began at 5 bar, with increasing its amount. The injection was effective to increase the CH4 content to 82.8%, 87.2%, and 90.6% at 0.09, 0.13, and 0.18 L H2/g CODFW.fed of H2 injection amount, respectively. At 0.25 L H2/g CODFW.fed, there was a further increase of CH4 content to 92.1%, but the MPY was dropped with pH increase to 8.7 with residual H2 being detected (4% in the biogas). Microbial community analysis showed the increased abundance of piezo-tolerant microbe with pressure increase, and direct interspecies electron transfer contributors after H2 injection. In conclusion, the integration of two approaches enabled to directly produce high calorific biogas (90% > CH4, 180 MJ/m3 biogas) from high-strength FW at the lowest requirement of H2 (0.18 L H2/g CODFW.fed) ever reported.

Microbial biotransformation of CH

Here, we report an analysis method for determining PHA (polyhydroxyalkanoates) contents and their monomer composition in microbial cells based on pyrolysis gas chromatography combined with mass spectrometry (Py-GC/MS). Various kinds of microbial cells accumulating different PHA contents and monomer compositions were prepared through the cultivation of Ralstonia eutropha and recombinant Escherichia coli. Py-GC/MS could analyse these samples in a short time without complicated pretreatment steps. Characteristic peaks such as 2-butenoic acid, 2-pentenoic acid, and hexadecanoic acid regarding PHA compositions and cell components were identified. Considering constituents of cells and ratios of peak areas of dehydrated monomers to hexadecanoic acid, a simple equation for estimation of PHA contents in microbial cells was derived. Also, monomer compositions of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in R. eutropha could be successfully determined based on peak area of 2-butenoic acid and 2-pentenoic acid of Py-GC/MS, which are the corresponding species of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in PHBV. Correlation of results between GC-FID and Py-GC/MS could be fitted very well. This method shows similar results for the samples obtained from same experimental conditions, allowing rapid and reliable analysis. Py-GC/MS can be a promising tool to rapidly screen PHA-positive strains based on polymer contents along with monomer compositions.

The microbial electrosynthesis is a platform to supply protons and electrons to improve the conversion efficiency and production rate for the valorization of C1 gas. This study examined proton migration and electron transfer of the electrode and microbe by using various external parameters in the electrosynthesis of CO. The CO electrosynthesis achieved almost double of coulombic efficiency than the conventional CO2 electrosynthesis. The maximum volumetric acetate production rate was 0.71 g/L/day in the BES, which was 2–6 times higher than reported elsewhere. These results show that the efficient proton migration and electron transfer can enhance the productivity and conversion efficiency of the biological CO conversion in a bioelectrochemical system.

The Two-component Regulatory System (TCS) is the primary mode that bacteria use to continuously sense the environment. A TCS is comprised of a periplasmic sensor Histidine kinase (HK) domain and a cytoplasmic Response regulator (RR) domain. The HK domain phosphorylates the RR domain to activate the effector gene expression. Utilizing a rational approach, the sensor HK was genetically engineered in Escherichia coli to create chimeric HK, by a rewiring or domain swapping strategy. Apart from the wild-type characteristics, chimeric HK imparts novel or the desired characteristics and ability to genetically engineered E. coli for its adaptation and survival. This review focuses on the design, potential applications, and future perspectives of chimeric HKs used as high throughput screening biosensors of various compounds.

Immuno-assay is one of diagnostic methods that usually measures biomarkers associated with cancers. However, this method is complex and take a long time to analyze. To overcome these disadvantages, many immuno-sensing chips have been designed and developed. However, these devices still require an external pump or electrical source. In this study, our group fabricated a capillary microfluidic device using glass and adhesive polyethylene terephthalate (PET) film, which were designed by simply patterning and cutting to make the microfluidic capillary channels. Using capillary force alone, glass microfluidic chip can control the speed of fluid-flow and the flow sequence by adjusting the width of the channel and design. In addition, each flow can push out other flow without mixing. The glass-based capillary microfluidic chip (GCMC) can automatically perform immunoassay in regular order without external devices and it provide an electrochemical signal analysis in an average of 2 min. The concentration of the prostate-specific antigen (PSA), a biomarker of prostate cancer, was measured by cyclic voltammetry (CV). In conclusion, GCMC can detect between a range of 100 pg/ml to 1 μg/ml of PSA and provide high selectivity to PSA.

Creating a module that achieves sustainable CO2 capture while being compatible with the existing industry is paramount in overcoming the current CO2-driven environmental issues. This paper presents...

World-wide production of l-lysine has rapidly increased in recent years. In the industrial scale production, it is cost effective to minimize waste as many waste materials are generated during downstream processing. Therefore, the conversion of crude lysine to a more valuable product reduces waste emission. In this study, 1,5-diaminopentane (DAP, trivial name: cadaverine) was produced by l-lysine decarboxylation using Hafnia alvei. The conditions of enzymatic reaction were determined. In particular, the addition of specific detergent (Brij 56) was significantly affected in the bioconversion system. Addition of hydrophobic organic solvent improved the mixing of the reactants. Finally, an industrial crude form of lysine served as a substrate. The DAP conversion by analytical, feed and industrial crude l-lysine was 93.9%, 90.3%, and 63.8%, respectively.

Here, we developed a pressurized bioreactor system that increase carbon monoxide (CO) transfer efficiency in order to enhance the hydrogen productivity in the microbial water gas shift reaction by Thermococcus onnurineus NA1. The effects of CO pressure on the hydrogen production rate, CO consumption rate and the cell growth were investigated using small scale stainless steel bottles at various CO partial pressures. It was found that CO solubility increased by applying pressure can affect hydrogen production positively as long as the increased toxicity of CO is endurable to cells. The hydrogen productivity increased to some extent with CO pressure, but decreased drastically at the pressure higher than 4 bar. On the other hand, the effect of pressure itself on the cell's activity was not as significant as that of CO solubility increase. In the experiments at various system pressures with identical CO partial pressure of 1 bar, more than 80% of the cell activity remains even at total pressure of 10 bar. Also, it was important to determine the appropriate time to increase pressure for preventing excess CO in the reactor. Based on these results, a fermentation strategy for the pressurized system was designed and applied to a 5 L bioreactor with the continuous supply of the gas containing 60% CO. When the pressure was introduced to the bioreactor up to 4 bar at CO limitation condition, the unprecedented high productivity (360 mmol L−1 h−1) could be obtained.

The purpose of this study was to simultaneously produce biodiesel and glycerol carbonate. Glycerol carbonate is a high-value material that could be obtained from glycerol. Response surface methodology was used to perform optimization of simultaneous enzymatic production of biodiesel and glycerol carbonate. Independent factors for optimization of simultaneous enzymatic production included enzyme concentration, a molar ratio of dimethyl carbonate to soybean oil, and reaction temperature. The levels of satisfaction with the production reaction model for biodiesel and glycerol carbonate were high, 0.994 and 1, respectively. The optimum condition for simultaneous production was 116.76 g/L enzyme concentration, 9.27:1 molar ratio, and 52.56 °C reaction temperature. Under the optimum condition, biodiesel 96.3% and glycerol carbonate 99.7% were produced at the same time.

The present study focused on the effect of reducibility of support oxide (CeO2, ZrO2, TiO2, and Al2O3) over the activity of cobalt-based catalysts in high temperature water-gas shift (HT-WGS) reaction. The H2 temperature programmed reduction (TPR) and CO chemisorption characterization results showed that dispersion of cobalt over the support was increased with the increase of the reducibility of the support oxide. The supported catalysts were characterized by X-ray diffraction (XRD) and N2 adsorption–desorption. The results indicated that the Co/CeO2 catalyst possesses the highest surface area and metal dispersion in the series; Co/CeO2 > Co/ZrO2 > Co/Al2O3 > Co/TiO2. The activity results showed that Co/CeO2 was the highly active among the tested catalysts in the temperature range of 350–550 °C. Moreover, the time-on-stream study revealed that the Co/CeO2 catalyst was relatively more stable than cobalt supported on ZrO2 and Al2O3 oxides. The excellent activity and stability of the Co/CeO2 catalyst were attributed to its high metal dispersion, which is found strongly dependent on the reducible nature of the support.

Coal is one of the major sources of energy, fuel, and other related chemicals. The processes to utilize coal for energy, fuel and other chemicals such as coal combustion, liquefaction, carbonization, and gasification pose a great threat to the environment by emitting toxic particles and CO2 to the atmosphere. Thus, biological beneficiation of coal can be a good strategy to utilize coal with environmental sustainability. Here, we report the screening of microorganisms able to degrade or depolymerize coal. These host strains are potential candidates for the development of biological treatment process of coal. A total of 45 microbial strains were isolated from sludge enriched with coal and were identified based on 16S rRNA sequencing. Four strains of three genera, Cupriavidus sp., Pseudomonas sp., and Alcaligenes sp., were further characterized for their abilities to degrade coal. The degree of coal degradation was analyzed by measuring the increase in absorbance at 450 nm by UV spectroscopy. These microorganisms were also able to increase the pH of the culture media as a response to the acidic nature of coal. Laccase-like activity was also found in these strains when tested for RBBR dye degradation. Since biological degradation of coal through the use of microorganisms is a good alternative to chemical combustion of coal, microbial strains isolated in this study can be potential biological catalysts for coal conversion into valuable chemicals.

Pyrolysis is an important conversion process which can produce high value-added light oils from unconventional oils such as oil sand bitumen and extra heavy oil, thus it is important to understand the characteristics and kinetics of pyrolysis for unconventional oils. In this study, the nonisothermal pyrolysis of Athabasca oil sand bitumen and its maltene and asphaltene fractions was analyzed using a thermogravimetric analyzer, and activation energies for pyrolysis were determined by the model-free isoconversional Friedman analysis. The analysis suggests that the pyrolysis of oil sand bitumen consists of reactions for volatilization of maltene fraction and cracking of maltene and asphaltene fractions. The pyrolysis behavior of oil sand bitumen was well described based on the kinetic parameters estimated by the distributed activation energy model for maltene and asphaltene fractions, which is beneficial to effective utilization and development of pyrolysis processes of oil sand bitumen.

Advances in scientific technology in the early twentieth century have facilitated the development of synthetic plastics that are lightweight, rigid, and can be easily molded into a desirable shape without changing their material properties. Thus, plastics become ubiquitous and indispensable materials that are used in various manufacturing sectors, including clothing, automotive, medical, and electronic industries. However, strong physical durability and chemical stability of synthetic plastics, most of which are produced from fossil fuels, hinder their complete degradation when they are improperly discarded after use. In addition, accumulated plastic wastes without degradation have caused severe environmental problems, such as microplastics pollution and plastic islands. Thus, the usage and production of plastics is not free from environmental pollution or resource depletion. In order to lessen the impact of climate change and reduce plastic pollution, it is necessary to understand and address the current plastic life cycles. In this review, "sustainable biopolymers" are suggested as a promising solution to the current plastic crisis. The desired properties of sustainable biopolymers and bio-based and bio/chemical hybrid technologies for the development of sustainable biopolymers are mainly discussed.

While the hydrogen economy is receiving growing attention, research on microbial hydrogen production is also increasing. Microbial water–gas shift reaction is advantageous as it produces hydrogen from by product gas including carbon monoxide (CO). However, CO solubility in water is the bottleneck of this process by low mass transfer. Thermococcus onnurineus NA1 strain can endure a high-pressure environment and can enhance hydrogen production in a pressurized reactor by increasing CO solubility. As CO causes cell toxicity, two important factors, pressure and input gas flow rate, should be considered for process control during cultivation. Hence, we employed different operational strategies for enhancing hydrogen production and obtained 577 mmol/L/h of hydrogen productivity. This is the highest hydrogen productivity reported to date from microbial water–gas shift reaction.

We engineered a type II methanotroph, Methylosinus trichosporium OB3b, for 3-hydroxypropionic acid (3HP) production by reconstructing malonyl-CoA pathway through heterologous expression of Chloroflexus aurantiacus malonyl-CoA reductase (MCR), a bifunctional enzyme. Two strategies were designed and implemented to increase the malonyl-CoA pool and thus, increase in 3HP production. First, we engineered the supply of malonyl-CoA precursors by overexpressing endogenous acetyl-CoA carboxylase (ACC), substantially enhancing the production of 3HP. Overexpression of biotin protein ligase (BPL) and malic enzyme (NADP+-ME) led to a ∼22.7% and ∼34.5% increase, respectively, in 3HP titer in ACC-overexpressing cells. Also, the acetyl-CoA carboxylation bypass route was reconstructed to improve 3HP productivity. Co-expression of methylmalonyl-CoA carboxyltransferase (MMC) of Propionibacterium freudenreichii and phosphoenolpyruvate carboxylase (PEPC), which provides the MMC precursor, further improved the 3HP titer. The highest 3HP production of 49 mg/L in the OB3b-MCRMP strain overexpressing MCR, MMC and PEPC resulted in a 2.4-fold improvement of titer compared with that in the only MCR-overexpressing strain. Finally, we could obtain 60.59 mg/L of 3HP in 42 h using the OB3b-MCRMP strain through bioreactor operation, with a 6.36-fold increase of volumetric productivity compared than that in the flask cultures. This work demonstrates metabolic engineering of type II methanotrophs, opening the door for using type II methanotrophs as cell factories for biochemical production along with mitigation of greenhouse gases.

The surface hydrophobicity of a microbial cell is known to be one of the important factors in its adhesion to an interface. To date, such property has been altered by either genetic modification or external pH, temperature, and nutrient control. Here we report a new strategy to engineer a microbial cell surface and discover the unique dynamic trapping of hydrophilic cells at an air/water interface via hydrophobicity switching. We demonstrate the surface transformation and hydrophobicity switching of Escherichia coli (E. coli) by metal nanoparticles. By employing real-time dark-field imaging, we directly observe that hydrophobic gold nanoparticle-coated E. coli, unlike its naked counterpart, is irreversibly trapped at the air/water interface because of elevated hydrophobicity. We show that our surface transformation method and resulting dynamic interfacial trapping can be generally extended to Gram-positive bateria, Gram-negative bacteria, and fungi. As the dynamic interfacial trapping allows the preconcentration of microbial cells, high intensity of scattering light, in-plane focusing, and near-field enhancement, we are able to directly quantify E. coli as low as 1.0 × 103 cells/ml by using a smartphone with an image analyzer. We also establish the identification of different microbial cells by the characteristic Raman transitions directly measured from the interfacially trapped cells.

Biological production of 2,3-butanediol (2,3-BDO), a C4 platform chemical, has been studied recently, but the high cost of separation and purification before chemical conversion is substantial. To overcome this obstacle, we have conducted a study to convert 2,3-BDO to mevalonate, a terpenoid intermediate, using recombinant Pseudomonas putida and this biological process won’t need the separation and purification process of 2,3-BDO. The production of mevalonate when 2,3-BDO was used as a substrate was 6.61 and 8.44 times higher than when glucose and glycerol were used as substrates under the same conditions, respectively. Lower aeration contributed to higher yields of mevalonate in otherwise identical conditions. The maximum mevalonate production on the shaking flask scale was about 2.21 g/L, in this study (product yield was 0.295, 27% of theoretical yield (1.10)). This study was the first successful attempt for mevalonate production by P. putida using 2,3-BDO as the sole carbon source and presented a new metabolic engineering tool and biological process for mevalonate synthesis.

Microbial biotransformation of CH4 gas has been attractive for the production of energy and high-value chemicals. However, insufficient supply of CH4 in a culture medium needs to be overcome for the efficient utilization of CH4. Here, we utilized cellulose nanocrystals coated with a tannic acid-Fe3+ complex (TA-Fe3+CNCs) as a medium component to enhance the gas-liquid mass-transfer performance. TA-Fe3+CNCs were well suspended in water without agglomeration, stabilized gas bubbles without coalescence, and increased the gas solubility by 20 % and the kLa value at a rapid inlet gas flow rate. Remarkably, the cell growth rate of Methylomonas sp. DH-1 as model CH4-utilizing bacteria improved with TA-Fe3+CNC concentration without any cytotoxic or antibacterial properties, resulting in higher metabolite production ability such as methanol, pyruvate, formate, and succinate. These results showed that TA-Fe3+CNCs could be utilized as a significant component in the culture medium applicable as a promising nanofluid for efficient CH4 microbial biotransformation.

Coal, the most primitive fossil fuel, has been exploited for ages, and reserves dictate the economies of many countries. Presently, most energy is generated by direct combustion, raising concerns over global warming. Biological pretreatment of fossil resources and generation of alternative green energy can address the environmental issues associated with global coal utilization. Biological coal treatment can produce industrially important chemicals and bio-methane by employing microorganisms able to depolymerize/degrade coal. This review discusses current advances in microbial coal conversion, such as the efforts made to comprehend microbial processes, significant outputs of coal conversion, principle components responsible for coal conversion, and factors affecting the biological processes to convert coal. Development of these biological processes can be a stepping stone for greener coal; however, integration of multidisciplinary technologies is needed to increase the efficiency of economic coal utilization and production of economically and industrially feasible biomethane.

The objective of this study was to investigate microbial communities in a continuous anaerobic phenol-degrading system using a next generation sequencing tool. The anaerobic granules adapted to phenol were first obtained by repeated-batch operation, which were then inoculated in an up-flow anaerobic sludge blanket reactor (UASB) operated at various organic loading rates (OLRs). Lag periods for both phenol degradation and CH4 production decreased as batch fermentation was repeated, indicating a progressive adaptation of the granules to phenol. In the UASB operation, the highest OLR handled was 6 kg COD/m3/d, in which the attained biogas production rate, phenol degradation, and CH4 contents were 2.1 m3/m3/d, 79.0%, and 75.3%, respectively. Syntrophorhabdus and Clostridium were found to be the dominant bacteria, whose sum occupied around 60% of total bacterial sequences. In particular, there was a significant increase in Syntrophorhabdus (39.2% of total bacterial sequences), known to degrade phenol to benzoate and subsequently to acetate and hydrogen in syntrophic association with a hydrogenotrophic methanogen. In terms of archaea, Methanosaeta (42.1% of total archaeal sequences), and Methanobacterium (24.5% of total archaeal sequences) became dominant as operation continued, which were negligible in the inoculum.

Background Methane is the major component of natural and shale gas. Methane can be converted into methanol via a bioprocess using methanotrophs, and methanol is a valuable chemical feedstock for the production of value-added chemicals. This work demonstrates highly effective bioconversion of methane to methanol using a newly isolated novel methanotroph, Methylomonas sp. DH-1. Results A novel methanotroph strain was isolated from activated sludge from a brewery plant and characterized using phylogenetic analysis, electron microscopy and chemotaxonomic analysis. This aerobic, Gram-negative, non-motile rod-shaped type I methanotroph was designated as Methylomonas sp. DH-1. The growth condition of Methylomonas sp. DH-1 and batch methane-to-methanol bioconversion conditions such as methane concentration, pH, biocatalyst loading, concentration of formate and MDH inhibitor were analyzed and optimized. Methanol was produced from methane with a 1.340 g L−1 titer, a 0.332 g L−1 h−1 volumetric conversion rate and a 0.0752 g g−1 cell h−1 specific methanol conversion rate. Conclusion It was demonstrated that isolation and application of a new methanotroph strain is a practical way of improving bioconversion efficiency in the conversion of methane to methanol. Moreover, one promising feature of Methylomonas sp. DH-1 for methanol production was its extremely high tolerance to methanol up to 7%(v/v), which is advantageous for high-titer methanol production. © 2016 Society of Chemical Industry

Ce0.6Zr0.4O2 supported transition metal (Me = Ni, Cu, Co, and Mo) catalysts have been investigated to screen for the catalytic activity and selectivity for deoxygenation reaction of oleic acid. Me-Ce0.6Zr0.4O2 catalysts were prepared by a co-precipitation method. Ni-Ce0.6Zr0.4O2 catalyst exhibited much higher oleic acid conversion, selectivity for C9 to C17 compounds, and oxygen removal efficiency than the others. This is mainly ascribed to the presence of free Ni species, synergy effects between Ni and Ce0.6Zr0.4O2, and the highest BET surface area.

In hydrogen production by methanol steam reforming reaction with microchannel reactor, Al2O3 thin film formed by atomic layer deposition (ALD) was introduced on the surface of microchannel reactor prior to the coating of catalyst particles. Methanol conversion rate and hydrogen production rate, increased in the presence of Al2O3 thin film. Over-view and cross-sectional scanning electron microscopy study showed that the adhesion between catalyst particles and the surface of microchannel reactor enhanced due to the presence of Al2O3 thin film. The improvement of hydrogen production rate inside the channels of microreactor mainly came from the stable fixation of catalyst particles on the surface of microchannels.