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Biomedical subjects

Brian H Davison

Publications and source records attributed to Brian H Davison.

9 recordsLinked to original sources

The path forward for biofuels and biomaterials.

Biomass represents an abundant carbon-neutral renewable resource for the production of bioenergy and biomaterials, and its enhanced use would address several societal needs. Advances in genetics, biotechnology, process chemistry, and engineering are leading to a new manufacturing concept for converting renewable biomass to valuable fuels and products, generally referred to as the biorefinery. The integration of agroenergy crops and biorefinery manufacturing technologies offers the potential for the development of sustainable biopower and biomaterials that will lead to a new manufacturing paradigm.

Biomass↗

Determination and comparison of the baseline proteomes of the versatile microbe Rhodopseudomonas palustris under its major metabolic states.

Rhodopseudomonas palustris is a purple nonsulfur anoxygenic phototrophic bacterium that is ubiquitous in soil and water. R. palustris is metabolically versatile with respect to energy generation and carbon and nitrogen metabolism. We have characterized and compared the baseline proteome of a R. palustris wild-type strain grown under six metabolic conditions. The methodology for proteome analysis involved protein fractionation by centrifugation, subsequent digestion with trypsin, and analysis of peptides by liquid chromatography coupled with tandem mass spectrometry. Using these methods, we identified 1664 proteins out of 4836 predicted proteins with conservative filtering constraints. A total of 107 novel hypothetical proteins and 218 conserved hypothetical proteins were detected. Qualitative analyses revealed over 311 proteins exhibiting marked differences between conditions, many of these being hypothetical or conserved hypothetical proteins showing strong correlations with different metabolic modes. For example, five proteins encoded by genes from a novel operon appeared only after anaerobic growth with no evidence of these proteins in extracts of aerobically grown cells. Proteins known to be associated with specialized growth states such as nitrogen fixation, photoautotrophic, or growth on benzoate, were observed to be up-regulated under those states.

Aerobiosis↗

Variation of S/G ratio and lignin content in a Populus family influences the release of xylose by dilute acid hydrolysis.

Wood samples from a second generation Populus cross were shown to have different lignin contents and S/G ratios (S: syringyl-like lignin structures; G: guaiacyl-like lignin structures). The lignin contents varied from 22.7% to 25.8% and the S/G ratio from 1.8 to 2.3. Selected samples spanning these ranges were hydrolyzed with dilute (1%) sulfuric acid to release fermentable sugars. The conditions were chosen for partial hydrolysis of the hemicellulosic fraction to maximize the expression of variation among samples. The results indicated that both lignin contents and S/G ratio significantly affected the yield of xylose. For example, the xylose yield of the 25.8% lignin and 2.3 S/G (high lignin, high S/G) sample produced 30% of the theoretical yield, whereas the xylose yield of the 22.7% lignin and 1.8 S/G (low lignin, low S/G) was 55% of the theoretical value. These results indicate that lignin content and composition among genetic variants within a single species can influence the hydrolyzability of the biomass.

Chemical Fractionation↗

Performance of chloroperoxidase stabilization in mesoporous sol-gel glass using in situ glucose oxidase peroxide generation.

A unique mesoporous sol-gel glass possessing a highly ordered porous structure (with three pore sizes of about 50, 150, and 200 A diameter) was used as a support material for immobilization of the enzyme chloroperoxidase (CPO). CPO was bound onto the glass via a bifunctional ligand, trimethoxysilylpropanal. In situ production of the cosubstrate, H2O2, was achieved using glucose oxidase. Solvent stability in acetonitrile mixtures was enhanced when a pore size larger than the size of CPO was used (i.e., 200 A). From these results, it appears that the glass-enzyme complex developed through the present work can be used as high-performance biocatalysts for various chemical-processing applications, particularly in harsh conditions.

Aldehydes↗

Succinic acid adsorption from fermentation broth and regeneration.

More than 25 sorbents were tested for uptake of succinic acid from aqueous solutions. The best resins were then tested for successive loading and regeneration using hot water. The key desired properties for an ideal sorbent are high capacity, complete stable regenerability, and specificity for the product. The best resins have a stable capacity of about 0.06 g of succinic acid/g of resin at moderate concentrations (1-5 g/L) of succinic acid. Several sorbents were tested more exhaustively for uptake of succinic acid and for successive loading and regeneration using hot water. One resin, XUS 40285, has a good stable isotherm capacity, prefers succinate over glucose, and has good capacities at both acidic and neutral pH. Succinic acid was removed from simulated media containing salts, succinic acid, acetic acid, and sugar using a packed column of sorbent resin, XUS 40285. The fermentation byproduct, acetate, was completely separated from succinate. A simple hot water regeneration successfully concentrated succinate from 10 g/L (inlet) to 40-110 g/L in the effluent. If successful, this would lower separation costs by reducing the need for chemicals for the initial purification step. Despite promising initial results of good capacity (0.06 g of succinic/g of sorbent), 70% recovery using hot water, and a recovered concentration of >100 g/L, this regeneration was not stable over 10 cycles in the column. Alternative regeneration schemes using acid and base were examined. Two (XUS 40285 and XFS-40422) showed both good stable capacities for succinic acid over 10 cycles and >95% recovery in a batch operation using a modified extraction procedure combining acid and hot water washes. These resins showed comparable results with actual broth.

Adsorption↗

A simplified method to create quantitative, "fixed" uranyl-contaminated metal coupons.

A method was developed and validated to quantitatively apply and "fix" uranyl contamination onto a metal surface (steel). Simple approaches are needed to create test surfaces in order to quantify contaminant removal or "decon" methods. We used steel discs sized to allow direct and accurate alpha counting in a Ludlum scanner from radioactive contaminants. A typical 3.8-cm-diameter coupon had a depleted uranyl loading of about 0.1 mg U cm with a count of 980 dpm. The resulting alpha radiation was measured with a precision of >97% for the same coupon. The alpha concentration on replicate coupons differed by as much as 9% (standard deviation). This method, based on earlier methods, required a uranyl solution to be dried but lowers the baking temperature to less than 100 degrees C to increase safety in a typical radiological laboratory. A dike was used to provide a uniform coating of the uranyl solution.

Calibration↗

Direct transesterification of gases by "dry" immobilized lipase.

Several different reactor configurations, including single pass, continuous recycle, and batch reactor modes, were used to investigate the effects of temperature and water activity, or relative humidity, on lipase-catalyzed, gas-phase transesterifications. Temperature and relative humidity were controlled both inside reactors and throughout the course of the reaction to account for and optimize their effects. Results indicated that, at low relative humidity, reaction rates increased with temperature up to 60 degrees C. However, when relative humidity was increased, a similar increase in temperature resulted in the loss of nearly all enzyme activity. These results are consistent with the idea that enzymes without free water are more thermally stable. Furthermore, at constant ambient temperatures, production increased dramatically with an increase in relative humidity, confirming the idea that an increase in water activity increases catalytic activity. A mass balance performed on reactors at higher relative humidity revealed that hydrolysis (rather than transesterification) of the ester substrate could significantly decrease product yields.

Animals↗

Substrate desolvation as a governing factor in enzymatic transformations of PAHs in aqueous-acetonitrile mixtures.

Conversion of hydrophobic substrates such as polyaromatic hydrocarbons (PAHs) was studied in aqueous-organic media using transformation of pyrene by cytochrome c. The experiments were conducted in pure solvents and aqueous-acetonitrile mixtures. The reaction rates dropped gradually as the solvent concentration was increased and were negligible in pure solvents. Thermodynamic calculations show that substrate desolvation was the governing factor in this reaction. The decrease in free energy of activation upon increasing the solvent concentration was found to be proportional to the substrate transfer free energy. This suggests that in order to make such reactions feasible in organic media, it will be necessary to improve the binding between the proteins and the substrates.

Acetonitriles↗

Characterization of the 70S Ribosome from Rhodopseudomonas palustris using an integrated "top-down" and "bottom-up" mass spectrometric approach.

We present a comprehensive mass spectrometric approach that integrates intact protein molecular mass measurement ("top-down") and proteolytic fragment identification ("bottom-up") to characterize the 70S ribosome from Rhodopseudomonas palustris. Forty-two intact protein identifications were obtained by the top-down approach and 53 out of the 54 orthologs to Escherichia coli ribosomal proteins were identified from bottom-up analysis. This integrated approach simplified the assignment of post-translational modifications by increasing the confidence of identifications, distinguishing between isoforms, and identifying the amino acid positions at which particular post-translational modifications occurred. Our combined mass spectrometry data also allowed us to check and validate the gene annotations for three ribosomal proteins predicted to possess extended C-termini. In particular, we identified a highly repetitive C-terminal "alanine tail" on L25. This type of low complexity sequence, common to eukaryotic proteins, has previously not been reported in prokaryotic proteins. To our knowledge, this is the most comprehensive protein complex analysis to date that integrates two MS techniques.

Acetylation↗