Single-well push-pull tests evaluating isobutane as a primary substrate for promoting in situ cometabolic biotransformation reactions.

A series of single-well push-pull tests (SWPPTs) were performed to investigate the efficacy of isobutane (2-methylpropane) as a primary substrate for in situ stimulation of microorganisms able to cometabolically transform common groundwater contaminants, such as chlorinated aliphatic hydrocarbons and 1,4-dioxane (1,4-D). In biostimulation tests, the disappearance of isobutane relative to a nonreactive bromide tracer indicated an isobutane-utilizing microbial community rapidly developed in the aquifer around the test well. SWPPTs were performed as natural drift tests with first-order rates of isobutane consumption ranging from 0.4 to 1.4 day-1. Because groundwater contaminants were not present at the demonstration site, isobutene (2-methylpropene) was used as a nontoxic surrogate to demonstrate cometabolic activity in the subsurface after biostimulation. The transformation of isobutene to isobutene epoxide (2-methyl-1,2-epoxypropane) illustrates the epoxidation process previously shown for common groundwater contaminants after cometabolic transformation by alkane-utilizing bacteria. The rate and extent of isobutene consumption and the formation and transformation of isobutene epoxide were greater in the presence of isobutane, with no evidence of primary substrate inhibition. Modeled concentrations of isobutane-utilizing biomass in microcosms constructed with groundwater collected before and after each SWPPT offered additional evidence that the isobutane-utilizing microbial community was stimulated in the aquifer. Experiments in groundwater microcosms also demonstrated that the isobutane-utilizing bacteria stimulated in the subsurface could cometabolically transform a mixture of co-substrates including isobutene, 1,1-dichloroethene, cis-1,2-dichloroethene, and 1,4-D with the same co-substrate preferences as the bacterium Rhodococcus rhodochrous ATCC strain 21198 after growth on isobutane. This study demonstrated the effectiveness of isobutane as primary substrate for stimulating in situ cometabolic activity and the use of isobutene as surrogate to investigate in situ cometabolic reactions catalyzed by isobutane-stimulated bacteria.

Hyperpolarized (129) Xe MRI using isobutene as a new quenching gas.

The use of a quenching gas, isobutene, with a low vapor pressure was investigated to enhance the utility of hyperpolarized (129) Xe (HP Xe) MRI. Xenon mixed with isobutene was hyperpolarized using a home-built apparatus for continuously producing HP Xe. The isobutene was then readily liquefied and separated almost totally by continuous condensation at about 173 K, because the vapor pressure of isobutene (0.247 kPa) is much lower than that of Xe (157 kPa). Finally, the neat Xe gas was continuously delivered to mice by spontaneous inhalation. The HP Xe MRI was enhanced twofold in polarization level and threefold in signal intensity when isobutene was adopted as the quenching gas instead of N2 . The usefulness of the HP Xe MRI was verified by application to pulmonary functional imaging of spontaneously breathing mice, where the parameters of fractional ventilation (ra ) and gas exchange (fD ) were evaluated, aiming at future extension to preclinical studies. This is the first application of isobutene as a quenching gas for HP Xe MRI.

Hierarchical SAPO-34 Catalysts as Host for Cu Active Sites.

Cu-containing hierarchical SAPO-34 catalysts were synthesized by the bottom-up method using different mesoporogen templates: CTAB encapsulated within ordered mesoporous silica nanoparticles (MSNs) and sucrose. A high fraction of the Cu centers exchanged in the hierarchical SAPO-34 architecture with high mesopore surface area and volume was achieved when CTAB was embedded within ordered mesoporous silica nanoparticles. Physicochemical characterization was performed by using structural and spectroscopic techniques to elucidate the properties of hierarchical SAPO-34 before and after Cu introduction. The speciation of the Cu sites, investigated by DR UV-Vis, and the results of the catalytic tests indicated that the synergy between the textural properties of the hierarchical SAPO-34 framework, the high Cu loading, and the coordination and localization of the Cu sites in the hierarchical architecture is the key point to obtaining good preliminary results in the NO selective catalytic reduction with hydrocarbons (HC-SCR).

Isobutylene contamination of blood collected in 10-ml evacuated blood collection tubes with gray conventional rubber stoppers.

Dual-column headspace gas chromatographic analysis with two flame-ionization detectors is a commonly used analytical technique for forensic blood ethanol quantitation. This technique is also applicable to the identification and quantitation of other volatile organic compounds such as methanol in biological samples. Compound identification by retention time is limited to those compounds with known retention times programmed into the instrument method. Historically, an early-eluting peak from an unidentified compound has been observed in both chromatograms from antemortem blood samples analyzed for ethanol concentration with this technique. The unidentified compound's retention time matches that of methanol on one column but not on the second column. This previously unidentified compound has been identified as isobutylene. The proposed source of the isobutylene contamination historically observed in antemortem blood samples collected in 10-ml gray-top blood collection tubes is the conventional rubber stopper. Isobutylene was detected in deionized water stored in each of the seven lots of 10-ml blood tubes tested; the expiration dates of the tubes tested spanned the years 2002-2022. Misidentification of isobutylene as methanol is possible when using a single-column gas chromatographic system. The presence of isobutylene in blood collected in a gray-top collection tube does not represent laboratory contamination, is not an interferent with blood ethanol quantitation, and does not affect the ethanol concentration in the blood. A 0.150 g/dl aqueous ethanol standard was stored in a gray-top tube to evaluate the potential impact of isobutylene on ethanol quantitation. The solution's average ethanol concentration measured after storage was 0.150 g/dl.

Isobutene production in Synechocystis sp. PCC 6803 by introducing α-ketoisocaproate dioxygenase from Rattus norvegicus.

Cyanobacteria can be utilized as a platform for direct phototrophic conversion of CO2 to produce several types of carbon-neutral biofuels. One promising compound to be produced photobiologically in cyanobacteria is isobutene. As a volatile compound, isobutene will quickly escape the cells without building up to toxic levels in growth medium or get caught in the membranes. Unlike liquid biofuels, gaseous isobutene may be collected from the headspace and thus avoid the costly extraction of a chemical from culture medium or from cells. Here we investigate a putative synthetic pathway for isobutene production suitable for a photoautotrophic host. First, we expressed α-ketoisocaproate dioxygenase from Rattus norvegicus (RnKICD) in Escherichia coli. We discovered isobutene formation with the purified RnKICD with the rate of 104.6 â€‹± â€‹9 â€‹ng (mg protein)-1 min-1 using α-ketoisocaproate as a substrate. We further demonstrate isobutene production in the cyanobacterium Synechocystis sp. PCC 6803 by introducing the RnKICD enzyme. Synechocystis strain heterologously expressing the RnKICD produced 91 â€‹ng â€‹l-1 OD750 -1 â€‹h-1. Thus, we demonstrate a novel sustainable platform for cyanobacterial production of an important building block chemical, isobutene. These results indicate that RnKICD can be used to further optimize the synthetic isobutene pathway by protein and metabolic engineering efforts.

Comparative Study of Different Acidic Surface Structures in Solid Catalysts Applied for the Isobutene Dimerization Reaction.

Dimerization of isobutene (IBE) to C8s olefins was evaluated over a range of solid acid catalysts of diverse nature, in a fixed bed reactor working in a continuous mode. All catalytic materials were studied in the title reaction performed between 50-250 °C, being the reaction feed a mixture of IBE/helium (4:1 molar ratio). In all materials, both conversion and selectivity increased with increasing reaction temperature and at 180 °C the best performance was recorded. Herein, we used thermogravimetry analysis (TGA) and temperature programmed desorption of adsorbed ammonia (NH3-TPD) for catalysts characterization. We place emphasis on the nature of acid sites that affect the catalytic performance. High selectivity to C8s was achieved with all catalysts. Nicely, the catalyst with higher loading of Brønsted sites displayed brilliant catalytic performance in the course of the reaction (high IBE conversion). However, optimum selectivity towards C8 compounds led to low catalyst stability, this being attributed to the combined effect between the nature of acidic sites and structural characteristics of the catalytic materials used. Therefore, this study would foment more research in the optimization of the activity and the selectivity for IBE dimerization reactions.

Actinobacterial Degradation of 2-Hydroxyisobutyric Acid Proceeds via Acetone and Formyl-CoA by Employing a Thiamine-Dependent Lyase Reaction.

The tertiary branched short-chain 2-hydroxyisobutyric acid (2-HIBA) has been associated with several metabolic diseases and lysine 2-hydroxyisobutyrylation seems to be a common eukaryotic as well as prokaryotic post-translational modification in proteins. In contrast, the underlying 2-HIBA metabolism has thus far only been detected in a few microorganisms, such as the betaproteobacterium Aquincola tertiaricarbonis L108 and the Bacillus group bacterium Kyrpidia tusciae DSM 2912. In these strains, 2-HIBA can be specifically activated to the corresponding CoA thioester by the 2-HIBA-CoA ligase (HCL) and is then isomerized to 3-hydroxybutyryl-CoA in a reversible and B12-dependent mutase reaction. Here, we demonstrate that the actinobacterial strain Actinomycetospora chiangmaiensis DSM 45062 degrades 2-HIBA and also its precursor 2-methylpropane-1,2-diol via acetone and formic acid by employing a thiamine pyrophosphate-dependent lyase. The corresponding gene is located directly upstream of hcl, which has previously been found only in operonic association with the 2-hydroxyisobutyryl-CoA mutase genes in other bacteria. Heterologous expression of the lyase gene from DSM 45062 in E. coli established a 2-hydroxyisobutyryl-CoA lyase activity in the latter. In line with this, analysis of the DSM 45062 proteome reveals a strong induction of the lyase-HCL gene cluster on 2-HIBA. Acetone is likely degraded via hydroxylation to acetol catalyzed by a MimABCD-related binuclear iron monooxygenase and formic acid appears to be oxidized to CO2 by selenium-dependent dehydrogenases. The presence of the lyase-HCL gene cluster in isoprene-degrading Rhodococcus strains and Pseudonocardia associated with tropical leafcutter ant species points to a role in degradation of biogenic short-chain ketones and highly branched organic compounds.

Efficient Separation of n-Butene and iso-Butene by Flexible Ultramicroporous Metal-Organic Frameworks with Pocket-like Cavities.

Efficient separation of n-butene (n-C4 H8 ) and iso-butene (iso-C4 H8 ) is of significance for the upgrading of C4 olefins to high-value end products but remains one of the major challenges in hydrocarbon purifications owing to their similar structures. Herein, we report a flexible metal-organic framework, MnINA (INA=isonicotinate), featuring one-dimensional pore channels with periodically large pocket-like cavities connected by narrow bottlenecks, for the first time for efficient n-/iso-C4 H8 separation. MnINA with smaller pore size (4.62 Å) compared with CuINA (4.84 Å), exhibits steep adsorption isotherms and high capacity of 1.79 mmol g-1 for n-C4 H8 (4.46 Å) through strong host-guest interactions via C-H⋅⋅⋅π bonding. The narrow bottlenecks exert barriers for the large molecules of iso-C4 H8 (4.84 Å) within the gate-opening pressure range of 0-0.1 bar. This gives rise to MnINA with excellent separation selectivity of 327.7 for n-/iso-C4 H8 mixture. The adsorption mechanism for n-C4 H8 and the gate-opening effect were investigated by dispersion-corrected density functional (DFT-D) theory, verifying the strong interactions between n-C4 H8 and the frameworks as well as the gate-opening effect derived from the rotation of organic linkers. The breakthrough tests confirmed MnINA and CuINA can be promising candidates for n-/iso-C4 H8 separation.

Hybrid Mo-C T Nanowires as Highly Efficient Catalysts for Direct Dehydrogenation of Isobutane.

Direct dehydrogenation of isobutane to isobutene has drawn extensive attention for synthesizing various chemicals. The Mo-based catalysts hold promise as an alternative to the toxic CrO x- and scarce Pt-based catalysts. However, the low activity and rapid deactivation of the Mo-based catalysts greatly hinder their practical applications. Herein, we demonstrate a feasible approach toward the development of efficient and non-noble metal dehydrogenation catalysts based on Mo-C T hybrid nanowires calcined at different temperatures. In particular, the optimal Mo-C700 catalyst exhibits isobutane consumption rate of 3.9 mmol g-1 h-1 and isobutene selectivity of 73% with production rate of 2.8 mmol g-1 h-1. The catalyst maintained 90% of its initial activity after 50 h of reaction. Extensive characterizations reveal that such prominent performance is well correlated with the adsorption abilities of isobutane and isobutene and the formation of η-MoC species. In contrast, the generation of ß-Mo2C crystalline phase during long-term reaction causes minor decline in activity. Compared to MoO2 and ß-Mo2C, η-MoC plays a role more likely in suppressing the cracking reaction. This work demonstrates a feasible approach toward the development of efficient and non-noble metal dehydrogenation catalysts.

Low Leachable Container System Consisting of a Polymer-Based Syringe with Chlorinated Isoprene Isobutene Rubber Plunger Stopper.

UNLABELLED: A 36 month leachable study on water for injection in direct contact within a polymer-based prefillable syringe consisting of a cyclo olefin polymer barrel, a chlorinated isoprene isobutene rubber plunger stopper, a polymer label attached on the barrel, and a secondary packaging was conducted at 25 ± 2 °C and 60 ± 5% relative humidity. Through the various comparison studies, no difference in the leachable amounts was observed between this polymer-based prefilled syringe and a glass bottle as a blank sample reference by 36 months. No influence on the leachables study outcome was noted from the printed label and/or label adhesive or from the secondary packaging. In an additional study, no acrylic acid used as the label adhesive leachable was detected by an extended storage for 45 months at 25 ± 2 °C and 60 ± 5% relative humidity as a worst case. To obtain more details, a comparison extractable study was conducted between a cyclo olefin polymer barrel and a glass barrel. In addition, chlorinated isoprene isobutene rubber and bromo isoprene isobutene rubber were compared. As a result, no remarkable difference was found in the organic extractables for syringe barrels. On the other hand, in the case of element extractable analysis, the values for the cyclo olefin polymer barrel were lower than that for the glass barrel. For the plunger stoppers, the chlorinated isoprene isobutene rubber applied in this study was showing a lower extractable profile as compared to the bromo isoprene isobutene rubber, both for organic and element extractables. In conclusion, the proposed polymer-based prefillable syringe system has great potential and represents a novel alternative that can achieve very low level extractable profiles and can bring additional value to the highly sensitive biotech drug market. LAY ABSTRACT: A 36 month leachable study on water for injection in direct contact within a cyclo olefin polymer barrel and chlorinated isoprene isobutene rubber plunger stopper that has a polymer label attached to the barrel and is wrapped into a secondary packaging was conducted at 25 °C and 60% relative humidity. Through the various comparison studies, no difference in the leachable amounts was observed between polymer-based prefilled syringes and a glass bottle as a blank sample reference by 36 months. No influences on the leachables study outcome were noted from the secondary packaging. To obtain more details, a comparison extractable study was conducted between the cyclo olefin polymer and the glass barrel. In addition, chlorinated isoprene isobutene rubber and bromo isoprene isobutene rubber plunger stoppers were compared as well. As a result, no remarkable difference was found in the organic extractables for barrels. As for element extractable analysis, the values for the cyclo olefin polymer barrel were lower than that for the glass barrel. For the plunger stoppers, the chlorinated isoprene isobutene rubber applied in this study was showing a lower extractable profile as compared to the bromo isoprene isobutene rubber, both for organic and element extractables. In conclusion, the proposed polymer-based prefillable syringe system has great potential and represents a novel alternative that can achieve very low level extractable profiles and can bring additional value to the highly sensitive biotech drug market.

Metal-Exchanged ß Zeolites as Catalysts for the Conversion of Acetone to Hydrocarbons.

Various metal-ß zeolites have been synthesized under similar ion-exchange conditions. During the exchange process, the nature and acid strength of the used cations modified the composition and textural properties as well as the Brönsted and Lewis acidity of the final materials. Zeolites exchanged with divalent cations showed a clear decrease of their surface Brönsted acidity and an increase of their Lewis acidity. All materials were active as catalysts for the transformation of acetone into hydrocarbons. Although the protonic zeolite was the most active in the acetone conversion (96.8% conversion), the metal-exchanged zeolites showed varied selectivities towards different products of the reaction. In particular, we found the Cu-ß to have a considerable selectivity towards the production of isobutene from acetone (over 31% yield compared to 7.5% of the protonic zeolite). We propose different reactions mechanisms in order to explain the final product distributions.