Hydrocarbon Substrates Research Articles

Resonance Raman spectroscopic studies of peroxo and hydroperoxo intermediates in lauric acid (LA)-bound cytochrome P450 119

Cytochromes P450 bind and cleave dioxygen to generate a potent intermediate compound I, capable of hydroxylating inert hydrocarbon substrates. Cytochrome P450 119, a bacterial cytochrome P450 that serves as a good model system for the study of the intermediate states in the P450 catalytic cycle. CYP119 is found in high temperature and sulfur rich environments. Though the natural substrate and redox partner are still unknown, a potential application of such thermophilic P450s is utilizing them as biocatalysts in biotechnological industry; e.g., the synthesis of organic compounds otherwise requiring hostile environments like extremes of pH or temperature. In the present work the oxygenated complex of this enzyme bound to lauric acid, a surrogate substrate known to have a good binding affinity, was studied by a combination of cryoradiolysis and resonance Raman spectroscopy, to trap and characterize active site structures of the key fleeting enzymatic intermediates, including the peroxo and hydroperoxo species.

Progress in Creation of a New Constant Medium for Hyperpolarized MRI

The MRI (Magnetic Resonance Imaging) has been used as one of the powerful tools for medical diagnoses. Its usefulness is, however, still restricted because of the low spatial resolution and long measuring time mainly due to the low NMR (Nuclear Magnetic Resonance) signals relative to the noise levels. To overcome these restrictions, we started developing a method to remarkably enlarge the NMR signals about 10 years ago. We employ a method to hyperpolarize the nuclei, where the “hyperpolarize” means to artificially generate the nuclear polarization by many orders (102~106) of magnitude higher than the ordinary NMR signals currently in use. The hyperpolarized MRI would enable us to provide images with much higher spatial resolution and shorter measuring time than ever. Several techniques for hyperpolarization have been put into practical use; the BF (Brute Force) method, PHIP (Parahydrogen Induced Polarization) method, Laser optical pumping, DNP (Dynamic Nuclear Polarization), and so on. The experimental study on the hyperpolarization of 3He by the BF method, and that of 19F in PFC (Perfluorocarbon, known as an artificial blood) by the PHIP method has gotten started. In the former method, we use an extremely low temperature realized by the Pomeranchuk cooling in combination with 3He/4He dilution refrigerator and high magnetic field. In the latter method, we use the hydrogenation of the parahydrogen in the unsaturated hydrocarbon substrate at room temperature. Very recently, we started developing a novel type of the hyperpolarized MRI named HMM (Hyperpolarized Metabolic MRI) hoping for the cancer diagnosis.

Inhibition of Ammonia Monooxygenase from Ammonia-Oxidizing Archaea by Linear and Aromatic Alkynes.

Ammonia monooxygenase (AMO) is a key nitrogen-transforming enzyme belonging to the same copper-dependent membrane monooxygenase family (CuMMO) as the particulate methane monooxygenase (pMMO). The AMO from ammonia-oxidizing archaea (AOA) is very divergent from both the AMO of ammonia-oxidizing bacteria (AOB) and the pMMO from methanotrophs, and little is known about the structure or substrate range of the archaeal AMO. This study compares inhibition by C2 to C8 linear 1-alkynes of AMO from two phylogenetically distinct strains of AOA, "Candidatus Nitrosocosmicus franklandus" C13 and "Candidatus Nitrosotalea sinensis" Nd2, with AMO from Nitrosomonas europaea and pMMO from Methylococcus capsulatus (Bath). An increased sensitivity of the archaeal AMO to short-chain-length alkynes (≤C5) appeared to be conserved across AOA lineages. Similarities in C2 to C8 alkyne inhibition profiles between AMO from AOA and pMMO from M. capsulatus suggested that the archaeal AMO has a narrower substrate range than N. europaea AMO. Inhibition of AMO from "Ca Nitrosocosmicus franklandus" and N. europaea by the aromatic alkyne phenylacetylene was also investigated. Kinetic data revealed that the mechanisms by which phenylacetylene inhibits "Ca Nitrosocosmicus franklandus" and N. europaea are different, indicating differences in the AMO active site between AOA and AOB. Phenylacetylene was found to be a specific and irreversible inhibitor of AMO from "Ca Nitrosocosmicus franklandus," and it does not compete with NH3 for binding at the active site.IMPORTANCE Archaeal and bacterial ammonia oxidizers (AOA and AOB, respectively) initiate nitrification by oxidizing ammonia to hydroxylamine, a reaction catalyzed by ammonia monooxygenase (AMO). AMO enzyme is difficult to purify in its active form, and its structure and biochemistry remain largely unexplored. The bacterial AMO and the closely related particulate methane monooxygenase (pMMO) have a broad range of hydrocarbon cooxidation substrates. This study provides insights into the AMO of previously unstudied archaeal genera, by comparing the response of the archaeal AMO, a bacterial AMO, and pMMO to inhibition by linear 1-alkynes and the aromatic alkyne, phenylacetylene. Reduced sensitivity to inhibition by larger alkynes suggests that the archaeal AMO has a narrower hydrocarbon substrate range than the bacterial AMO, as previously reported for other genera of AOA. Phenylacetylene inhibited the archaeal and bacterial AMOs at different thresholds and by different mechanisms of inhibition, highlighting structural differences between the two forms of monooxygenase.

Characterisation of bubble diameter and gas hold-up in simulated hydrocarbon-based bioprocesses in a bubble column reactor

Hydrocarbon substrates can be upgraded to high-value products through biological oxidation processes, whereby an oxygen moiety is inserted into the hydrocarbon backbone by microbes in a suitable bioreactor system such as a bubble column reactor (BCR). However, there is a need to understand the behaviour of the Sauter bubble diameter (D32) and gas hold-up (ƐG) in a simulated four-phase (air, water, hydrocarbon and microbes) system in the BCR. This study has investigated the impact of operational conditions, such as hydrocarbon concentration (HC), superficial gas velocity (UG) and yeast loading (SL) (using deactivated S. cerevisiae as test microorganism) on D32 and ƐG. It was found that D32 and ƐG were mainly affected by UG and SL, whereas HC had an insignificant impact on both D32 and ƐG. Any increase in UG (1–3 cm/sec) resulted in a significant increase in D32 and ƐG, due to the increase in the number of bubbles in the system. On the other hand, an increase in SL was found to result in D32 linearly increasing, which thereafter caused a decrease of ƐG in the system. This influence can be attributed to the yeast cells in the system affecting the fluid properties and the system hydrodynamics. The outcome of this work provides fundamental understanding of the impact of operating conditions (HC, UG and SL) on D32 and ƐG, which underpins the system hydrodynamics in a simulated four-phase hydrocarbon-based bioprocess.

Enantioselective Bioamination of Aromatic Alkanes Using Ammonia: A Multienzymatic Cascade Approach

AbstractChiral amines are common drug building blocks and important active pharmaceutical ingredients. Preparing these functionalized compounds from simple materials, such as alkanes, is of great interest. We recently developed an artificial bioamination cascade for the C−H amination of cyclic alkanes by combining P450 monooxygenase, alcohol dehydrogenase, and amine dehydrogenase. Herein, this system has been extended to the synthesis of chiral aromatic amines. In the first hydroxylation step, process optimization increased the conversion to 77 %. Two stereoselectively complementary alcohol dehydrogenases and an amine dehydrogenase were selected for the bioconversion of aromatic hydrocarbons to amines. The amination reaction was optimized with respect to cofactor addition and enzyme dosage. Isopropanol was added to decrease ketone intermediate accumulation in the amination step, which further enhanced the overall conversion. This cascade system converted a panel of hydrocarbon substrates into the corresponding amines with excellent optical purity (>99 % ee) and moderate conversion ratios (13–53 %).

Microbial Degradation of Different Hydrocarbon Fuels with Mycoremediation of Volatiles.

Naturally occurring microorganisms in soil matrices play a significant role in overall hydrocarbon contaminant removal. Bacterial and fungal degradation processes are major contributors to aerobic remediation of surface contaminants. This study investigated degradation of conventional diesel, heating diesel fuel, synthetic diesel (Syntroleum), fish biodiesel and a 20% biodiesel/diesel blend by naturally present microbial communities in laboratory microcosms under favorable environmental conditions. Visible fungal remediation was observed with Syntroleum and fish biodiesel contaminated samples, which also showed the highest total hydrocarbon mineralization (>48%) during the first 28 days of the experiment. Heating diesel and conventional diesel fuels showed the lowest total hydrocarbon mineralization with 18–23% under favorable conditions. In concurrent experiments with growth of fungi suspended on a grid in the air space above a specific fuel with little or no soil, fungi were able to survive and grow solely on volatile hydrocarbon compounds as a carbon source. These setups involved negligible bacterial degradation for all five investigated fuel types. Fungal species able to grow on specific hydrocarbon substrates were identified as belonging to the genera of Giberella, Mortierella, Fusarium, Trichoderma, and Penicillium.

High-Valent d7 NiIII versus d8 CuIII Oxidants in PCET.

Oxygenases have been postulated to utilize d4 FeIV and d8 CuIII oxidants in proton-coupled electron transfer (PCET) hydrocarbon oxidation. In order to explore the influence the metal ion and d-electron count can hold over the PCET reactivity, two metastable high-valent metal-oxygen adducts, [NiIII(OAc)(L)] (1b) and [CuIII(OAc)(L)] (2b), L = N,N'-(2,6-diisopropylphenyl)-2,6-pyridinedicarboxamidate, were prepared from their low-valent precursors [NiII(OAc)(L)]- (1a) and [CuII(OAc)(L)]- (2a). The complexes 1a/b-2a/b were characterized using nuclear magnetic resonance, Fourier transform infrared, electron paramagnetic resonance, X-ray diffraction, and absorption spectroscopies and mass spectrometry. Both complexes were capable of activating substrates through a concerted PCET mechanism (hydrogen atom transfer, HAT, or concerted proton and electron transfer, CPET). The reactivity of 1b and 2b toward a series of para-substituted 2,6-di-tert-butylphenols (p-X-2,6-DTBP; X = OCH3, C(CH3)3, CH3, H, Br, CN, NO2) was studied, showing similar rates of reaction for both complexes. In the oxidation of xanthene, the d8 CuIII oxidant displayed a small increase in the rate constant compared to that of the d7 NiIII oxidant. The d8 CuIII oxidant was capable of oxidizing a large family of hydrocarbon substrates with bond dissociation enthalpy (BDEC-H) values up to 90 kcal/mol. It was previously observed that exchanging the ancillary anionic donor ligand in such complexes resulted in a 20-fold enhancement in the rate constant, an observation that is further enforced by comparison of 1b and 2b to the literature precedents. In contrast, we observed only minor differences in the rate constants upon comparing 1b to 2b. It was thus concluded that in this case the metal ion has a minor impact, while the ancillary donor ligand yields more kinetic control over HAT/CPET oxidation.

Solvent-Free Melting-Assisted Pyrolysis Strategy Applied on the Co/N Codoped Porous Carbon Catalyst

From the current demand for aromatic ketones, economical and environmentally friendly renewable catalysts are highly economical for the efficient selection of aralkyl oxides to form the corresponding aromatic ketone compounds. Yet, since the preparation of the Co–N–C catalyst is complicated at present, it is difficult to be widely applied to the oxidation of arylalkanes. In this study, a one-step solvent-free melting-assisted heat-treatment strategy is adopted to synthesize cobalt and nitrogen codoped carbon catalysts from sucrose, 5-(4-carboxyphenyl)-10,15,20-triphenylcobalt porphyrin (CoCPTPP), and KIT-6. Thanks to the interaction between the silicon hydroxyl of KIT-6 and the carboxyl hydroxyl of CoCPTPP as well as the distinctive properties of CoCPTPP with Co–N4 structures, the Co species in the as-synthesized catalysts are uniformly dispersed on carbon materials. Hereafter, the catalyst showed a high efficiency for the oxidation of arylalkane, e.g., the conversion rate of ethylbenzene is 96% and the selectivity of acetophenone is 99%. According to the characterizations including transmission electron microscopy etc, the results show that a porous special structure and thriving Co–Nx active sites in the catalysts is the main reason for the catalyst’s good catalytic activity and stability. In addition, the catalyst can also be used for the oxidation reaction of various aromatic hydrocarbon substrates with high catalytic activity.

Expression, purification, characterization and in silico analysis of newly isolated hydrocarbon degrading bleomycin resistance dioxygenase.

In the present investigation, we report cloning, expression, purification and characterization of a novel Bleomycin Resistance Dioxygenase (BRPD). His-tagged fusion protein was purified to homogeneity using Ni-NTA affinity chromatography, yielding 1.2mg of BRPD with specific activity of 6.25 Umg-1 from 600ml of E. coli culture. Purified enzyme was a dimer with molecular weight ~26kDa in SDS-PAGE and ~73kDa in native PAGE analysis. The protein catalyzed breakdown of hydrocarbon substrates, including catechol and hydroquinone, in the presence of metal ions, as characterized via spectrophotometric analysis of the enzymatic reactions. Bleomycin binding was proven using the EMSA gel retardation assay, and the putative bleomycin binding site was further determined by in silico analysis. Molecular dynamic simulations revealed that BRPD attains octahedral configuration in the presence of Fe2+ ion, forming six co-ordinate complexes to degrade hydroquinone-like molecules. In contrary, in the presence of Zn2+ ion BRPD adopts tetrahedral configuration, which enables degradation of catechol-like molecules.

A closed cycle for esterifying aromatic hydrocarbons with CO2 and alcohol.

The ability to functionalize hydrocarbons with CO2 could create opportunities for high-volume CO2 utilization. However, current methods to form carbon-carbon bonds between hydrocarbons and CO2 require stoichiometric consumption of very resource-intensive reagents to overcome the low reactivity of these substrates. Here, we report a simple semi-continuous cycle that converts aromatic hydrocarbons, CO2 and alcohol into aromatic esters without consumption of stoichiometric reagents. Our strategy centres on the use of solid bases composed of an alkali carbonate (M2CO3, where M+ = K+ or Cs+) dispersed over a mesoporous support. Nanoscale confinement disrupts the crystallinity of M2CO3 and engenders strong base reactivity at intermediate temperatures. The overall cycle involves two distinct steps: (1) CO32--promoted C-H carboxylation, in which the hydrocarbon substrate is deprotonated by the supported M2CO3 and reacts with CO2 to form a supported carboxylate (RCO2M); and (2) methylation, in which RCO2M reacts with methanol and CO2 to form an isolable methyl ester with concomitant regeneration of M2CO3.

Highly Selective and Catalytic Oxygenations of C−H and C=C Bonds by a Mononuclear Nonheme High‐Spin Iron(III)‐Alkylperoxo Species

The reactivity of a mononuclear high-spin iron(III)-alkylperoxo intermediate [FeIII (t-BuLUrea )(OOCm)(OH2 )]2+ (2), generated from [FeII (t-BuLUrea )(H2 O)(OTf)](OTf) (1) [t-BuLUrea =1,1'-(((pyridin-2-ylmethyl)azanediyl)bis(ethane-2,1-diyl))bis(3-(tert-butyl)urea), OTf=trifluoromethanesulfonate] with cumyl hydroperoxide (CmOOH), toward the C-H and C=C bonds of hydrocarbons is reported. 2 oxygenates the strong C-H bonds of aliphatic substrates with high chemo- and stereoselectivity in the presence of 2,6-lutidine. While 2 itself is a sluggish oxidant, 2,6-lutidine assists the heterolytic O-O bond cleavage of the metal-bound alkylperoxo, giving rise to a reactive metal-based oxidant. The roles of the urea groups on the supporting ligand, and of the base, in directing the selective and catalytic oxygenation of hydrocarbon substrates by 2 are discussed.

Highly Selective and Catalytic Oxygenations of C−H and C=C Bonds by a Mononuclear Nonheme High‐Spin Iron(III)‐Alkylperoxo Species

AbstractThe reactivity of a mononuclear high‐spin iron(III)‐alkylperoxo intermediate [FeIII(t‐BuLUrea)(OOCm)(OH2)]2+(2), generated from [FeII(t‐BuLUrea)(H2O)(OTf)](OTf) (1) [t‐BuLUrea=1,1′‐(((pyridin‐2‐ylmethyl)azanediyl)bis(ethane‐2,1‐diyl))bis(3‐(tert‐butyl)urea), OTf=trifluoromethanesulfonate] with cumyl hydroperoxide (CmOOH), toward the C−H and C=C bonds of hydrocarbons is reported.2oxygenates the strong C−H bonds of aliphatic substrates with high chemo‐ and stereoselectivity in the presence of 2,6‐lutidine. While2itself is a sluggish oxidant, 2,6‐lutidine assists the heterolytic O−O bond cleavage of the metal‐bound alkylperoxo, giving rise to a reactive metal‐based oxidant. The roles of the urea groups on the supporting ligand, and of the base, in directing the selective and catalytic oxygenation of hydrocarbon substrates by2are discussed.

Beneath the surface: Evolution of methane activity in the bacterial multicomponent monooxygenases

The bacterial multicomponent monooxygenase (BMM) family has evolved to oxidise a wide array of hydrocarbon substrates of importance to environmental emissions and biotechnology: foremost amongst these is methane, which requires among the most powerful oxidant in biology to activate. To understand how the BMM evolved methane oxidation activity, we investigated the changes in the enzyme family at different levels: operonic, phylogenetic analysis of the catalytic hydroxylase, subunit or folding factor presence, and sequence-function analysis across the entirety of the BMM phylogeny. Our results show that the BMM evolution of new activities was enabled by incremental increases in oxidative power of the active site, and these occur in multiple branches of the hydroxylase phylogenetic tree. While the hydroxylase primary sequence changes that resulted in increased oxidative power of the enzyme appear to be minor, the principle evolutionary advances enabling methane activity occurred in the other components of the BMM complex and in the recruitment of stability proteins. We propose that enzyme assembly and stabilization factors have independently-evolved multiple times in the BMM family to support enzymes that oxidise increasingly difficult substrates. Herein, we show an important example of evolution of catalytic function where modifications to the active site and substrate accessibility, which are the usual focus of enzyme evolution, are overshadowed by broader scale changes to structural stabilization and non-catalytic unit development. Retracing macroscale changes during enzyme evolution, as demonstrated here, should find ready application to other enzyme systems and in protein design.

Formation of a Wave Structure on the Surface of a Graphene Film

A wavy structure of the surface of graphene nanoparticles on a hydrocarbon substrate has been obtained. It has been shown that the wavy surface is the manifestation of the Kelvin-Helmholtz instability at the interface between liquid paraffin and graphene suspension with the inhomogeneity length λ = 250 nm. The manifestation of such wavy structures is due primarily to van der Waals forces.

The potential of indigenous bacteria from oil sludge for biosurfactant production using hydrolysate of agricultural waste

Abstract. Ni’matuzahroh, Sari SK, Ningrum IP, Pusfita AD, Marjayandari L, Trikurniadewi N, Ibrahim SNMM, Fatimah, Nurhariyati T, Surtiningsih T, Yuliani H. 2019. The potential of indigenous bacteria from oil sludge for biosurfactant production using hydrolysate of agricultural waste. Biodiversitas 20: 1374-1379. Biosurfactants are amphipathic compounds which are useful in various fields of health, industry, and remediation. Biosurfactants are produced by bacteria that grow in hydrocarbon or sugar substrates. Hydrolysis product of agricultural waste can be used as a biosurfactant production medium. This research aims to obtain biosurfactant producing bacteria from Balongan oil sludge, Indonesia. The ability to grow and produce biosurfactant by indigenous bacteria was tested using a medium of Synthetic Mineral Water (SMW) added by 209.3 ppm of rice straw hydrolysis product (RSHP). The growth of bacteria was evaluated through Total Plate Count (TPC) and biosurfactant production was evaluated through measurement of emulsification activity and surface tension. Six indigenous bacteria were capable to produce biosurfactants in the RSHP. Emulsification activity was not detected, but surface tension reduction was founded. The best biosurfactant was indicated by surface tension value of 53.56 mN/m with TPC value of 20.07 CFU/mL at the 5th day of incubation by BP (1) 5. The indigenous bacteria were identified as Propionibacterium BP (1) 1, Propionibacterium BP (1) 3, Bacillus BP (1) 4, Corynebacterium BP (1) 5, Corynebacterium BP (1) 8, and Rothia BP (1) 6. Utilization of sugar as hydrolysis product of agricultural waste is an innovation of raw materials for biosurfactant production.

Novel polyolefin based alkaline polymer electrolyte membrane for vanadium redox flow batteries

Low conductivity and stability of anion exchange membranes limited the durability and performance of corresponding fuel cells despite their unique advantages in compared to the proton exchange membrane fuel cells. In this work, we present a simple and efficient method to fabricate highly durable anion exchange membranes from commercial porous hydrocarbon substrates. Polyethylene/polypropylene (PE/PP) microfibers is employed as a substrate and radiation induced emulsion grafting is utilized to introduce poly(vinylbenzyl chloride) side chains. The grafted substrates with flexible content of side chains are converted to anion exchange membranes by converting into dense membranes and introducing quaternary amine groups. The dense membranes are characterized and employed in vanadium redox flow battery (VRFB) application. Developed membranes demonstrate high anion conductivity, low vanadium permeability and good chemical and mechanical stability. In compare to Nafion 117, superior performance is achieved for the developed membranes as the undesired vanadium ion permeability is reduced by 16 times and the energy efficiency significantly enhanced by 9% at current density of 80 mA cm−2. Such enhanced performance of the developed membrane is attributed to the presence of positive charged ionic groups in the membrane structure.

Production and characterization of lipopeptide biosurfactant producing Paenibacillus sp. D9 and its biodegradation of diesel fuel

The current research aimed at analyzing the biodegradation efficiency of a potent biosurfactant producing Paenibacillus sp. D9, along with the characterization of the surface-active compound produced during diesel fuel biodegradation. The biosurfactant production by Paenibacillus sp. D9 was evaluated using diesel fuel as culture medium, subsequently analyzed for its structural characteristics using different methods and thus determining the biodegradation utilization. This strain showed wide cell surface hydrophobicity against varieties of hydrocarbon substrate tested. Paenibacillus sp. D9 displayed higher hydrophobicity to the long-chain hydrocarbons mixtures tested such as 71.5% diesel fuel, 70.0% engine oil and 76.0% n-paraffin. The optimum condition for biosurfactant synthesis was obtained in a medium containing 10% (v/v) diesel fuel with a production yield of 4.7 g/L. The resultant biosurfactant reduced surface tension from 71.4 to 30.1 mN/m against carbon source utilized. The critical micelle concentration value of the biosurfactant was 200 mg/L with emulsification efficiencies against a wide range of hydrophobic pollutants. With different physiochemical and analytical methods, the study demonstrated that the genus Paenibacillus produced a low molecular weight lipopeptide biosurfactant. Its emulsifying ability further supports its potential use in environmental and biotechnological applications.

Anaerobic degradation of hexadecane and phenanthrene coupled to sulfate reduction by enriched consortia from northern Gulf of Mexico seafloor sediment

To advance understanding of the fate of hydrocarbons released from the Deepwater Horizon oil spill and deposited in marine sediments, this study characterized the microbial populations capable of anaerobic hydrocarbon degradation coupled with sulfate reduction in non-seep sediments of the northern Gulf of Mexico. Anaerobic, sediment-free enrichment cultures were obtained with either hexadecane or phenanthrene as sole carbon source and sulfate as a terminal electron acceptor. Phylogenetic analysis revealed that enriched microbial populations differed by hydrocarbon substrate, with abundant SSU rRNA gene amplicon sequences from hexadecane cultures showing high sequence identity (up to 98%) to Desulfatibacillum alkenivorans (family Desulfobacteraceae), while phenanthrene-enriched populations were most closely related to Desulfatiglans spp. (up to 95% sequence identity; family Desulfarculaceae). Assuming complete oxidation to CO2, observed stoichiometric ratios closely resembled the theoretical ratios of 12.25:1 for hexadecane and 8.25:1 for phenanthrene degradation coupled to sulfate reduction. Phenanthrene carboxylic acid was detected in the phenanthrene-degrading enrichment cultures, providing evidence to indicate carboxylation as an activation mechanism for phenanthrene degradation. Metagenome-assembled genomes (MAGs) revealed that phenanthrene degradation is likely mediated by novel genera or families of sulfate-reducing bacteria along with their fermentative syntrophic partners, and candidate genes linked to the degradation of aromatic hydrocarbons were detected for future study.

Updating the Chemiluminescence Oxygen-Aftereffect Method for Determining the Rate Constant of the Peroxy-Radical Self-Reaction: Oxidation of Cyclohexene.

Updating the facile chemiluminescence oxygen-aftereffect method, most suitable for determining the rate constant (kt ) of the peroxy-radical self-reaction (main chemiluminescence channel), pertained to considering the sensitivity of such a method toward a disturbing influence of the peroxy radicals of the initiator of the chain oxidation process. Such a disturbance may derive from the side chemiluminescent reaction, which involves peroxy radicals of both hydrocarbon and initiator. To examine the applicability and limitations of the chemiluminescence method under present scrutiny, cyclohexene was used as the model oxidizable hydrocarbon substrate. Computer simulations of the reaction and chemiluminescence kinetics have demonstrated the validity of the considered methodology at the value of the rate constant of the propagation of the overall chain process by peroxy radicals of the initiator higher than 1 m-1 s-1 . Despite that the chemiluminescence time profile and the stationary level of the total chemiluminescence intensity depend on the kinetics of the side chemiluminescence channel and on the ratio of the excited-state generation yields in the mentioned reaction channel and in the main chemiluminescence process, the value of kt assessed by the oxygen-aftereffect method has been found independent of variation of these characteristics.

Assembling a genome for novel nitrogen-fixing bacteria with capabilities for utilization of aromatic hydrocarbons

Metagenome from refinery wastewater treatment plant running under nitrogen stress was analyzed for mining of novel aromatic hydrocarbon-degrading bacteria. The sequence data were assembled using metaspade followed by binning using the Metabat tool to assemble genome; where coverage and depth were calculated using bowtie and samtools. The analysis picked a novel genome belonging to family Bradyrhizobiaceae, identified based on 16S rDNA gene which was supported by CheckM and Kraken analysis. Using RAST, the assembled genome showed the capabilities for nitrogen fixation with the utilization of multiple hydrocarbon substrates with 14 different types of oxygenases as mapped by Minpath. An additional genetic feature like genes for stress and resistance towards heavy metals and antibiotic suggested that the genome has gone through the rigorous process of adaptation. If such bacteria could be cultivated then it will open the broad window of bioremediation strategies under nitrogen stress environment.