Judith Jervis

The effect of overliming on the toxicity of dilute acid pretreated lignocellulosics: the role of inorganics, uronic acids and ether-soluble organics.

Although the treatment of dilute acid pretreated lignocellulosics with calcium hydroxide or carbonate (overliming) is known to improve the fermentability of carbohydrate-rich hydrolyzate streams, a firm understanding of the chemistry behind the process is lacking. Quantitative evaluation of inorganics, uronic acids, and non-polar organics indicates that only a portion of the improvement can be ascribed to these materials. Upon overliming the concentrations of inorganics either increase (Ca, Mg), decrease (Fe, P, Zn, K) or remain relatively the same (Al, Na). Furthermore, organic compounds that are not extractable with tert-butyl methyl ether (MTBE) are toxic to Zymomonas mobilis CP4(pZB5). Overliming and direct neutralization are somewhat effective in removing sulfate anions, although sulfate toxicity is considerably less than that of acetic acid. Uronic acids were found to be non-toxic under pH controlled conditions.

Identification of inhibitory components toxic toward Zymomonas mobilis CP4(pZB5) xylose fermentation

Zymomonas mobilis CP4(pZB5) is a recombinant bacterium that can produce ethanol from both xylose and glucose. The ethanol-producing efficiency of this organism is substantially impeded by toxic substances present in pretreated hydrolyzates or solid biomass substrates. Acetic acid and furfural (a pentose degradation product) are highly toxic to this organism at levels envisioned for a pretreated-hardwood liquid hydrolyzate. In addition, lignin degradation products and 5-hydroxymethylfurfural (a hexose degradation product) have a moderately toxic effect on the organism. Of the compounds studied, organic acids and aldehydes were found to be more inhibitory than lignin acids or the one alkaloid studied. Acetone:water and methanol extracts of solid biomass samples from red oak, white oak, and yellow poplar are toxic toZymomonas cell growth and ethanol production, with the extracts from white oak being the most toxic.

C-Glycosidic ellagitannins from white oak heartwood and callus tissues

Abstract Efforts were made to fully evaluate the performances of the two commonly used solvents, aq. Me 2 CO and aq. MeOH, on extraction of vescalagin and castalagin from freshly harvested white oak ( Quercus alba ). The results show that aq. Me 2 CO is superior to aq. MeOH in obtaining higher vescalagin and castalagin yields with lower total insoluble ellagitannins. A comparison of aq. Me 2 CO extracts from Q. alba heartwood and callus tissues reveals similar ellagitannin profiles with callus tissues producing almost 3 times as much castalagin and vescalagin via manipulation of the media nitrogen and copper concentrations.

Toxicity of hardwood extractives toward Saccharomyces cerevisiae glucose fermentation.

The toxicity of oak and yellow-poplar wood extracts, as well as a first-stage hardwood hydrolyzate liquor prepared from a red oak:white oak:yellow-poplar (1:1:1) sawdust, on Saccharomyces cerevisiae D5A was examined. Acetone/water and hot methanol extracts of solid biomass samples from white oak, red oak, and yellow-poplar gave 88-94% of the ethanol produced with controls. The organism was tolerant to the compounds present in the xylose-rich hydrolyzate, with fermentation efficiency being improved to 97% of that obtained with controls by using an overliming/thermal conditioning protocol.

The exo-proteome and exo-metabolome of Nostoc punctiforme (Cyanobacteria) in the presence and absence of nitrate.

Abstract The ability of nitrogen-fixing filamentous Cyanobacteria to adapt to multiple environments comes in part from assessing and responding to external stimuli, an event that is initiated in the extracellular milieu. While it is known that these organisms produce numerous extracellular substances, little work has been done to characterize both the metabolites and proteins present under standard laboratory growth conditions. We have assessed the extracellular milieu of Nostoc punctiforme when grown in liquid culture in the presence and absence of a nitrogen source (nitrate). The extracellular proteins identified were enriched in integrin β-propellor domains and calcium-binding sites with sequences unique to N. punctiforme, supporting a role for extracellular proteins in modulating species-specific recognition and behavior processes. Extracellular proteases are present and active under both conditions, with the cells grown with nitrate having a higher activity when normalized to chlorophyll levels. The released metabolites are enriched in peptidoglycan-derived tetrasaccharides, with higher levels in nitrate-free media.

Mass Spectral Analyses of Corn Stover Prehydrolysates To Assess Conditioning Processes

Flow injection electrospray (FIE) and LC-tandem mass spectrometry techniques were used to characterize corn stover acid hydrolysates before and after overliming and ammonia conditioning steps. Analyses were performed on samples without fractionation (dilution only) in an effort provide an inventory of ionizable substances. Statistical evaluation of the results indicates that the ammonia-treated and crude hydrolysates were more similar to one another than any other pairing, with conditioning leading to a decrease in malate levels. LC-tandem mass spectrometry studies were also developed to characterize the oligosaccharides present in each hydrolysate utilizing a hydrophilic interaction chromatographic separation method. Neutral and acidic pentose-based oligosaccharides (xylodextrins) with degrees of polymerization between 2 and 5 were quantified with 4-O-methyl glucuronic acid-containing dimer and trimers predominating. Conditioning had little effect on the quantified oligosaccharide pool.

Toward Understanding Monomeric Ellagitannin Biosynthesis

Ellagitannins are found in approximately 40 percent of all dicotyledenous plants ranging from raspberries to the oaks.1–3 By definition, this class of compounds is composed of only glucose and biaryl-linked gallic acid. Though these two compounds are relatively simple in structure, the complexity of the compounds that can be formed is tremendous. Ellagitannins are seemingly a natural combinatorial library, with over 500 different compounds isolated and identified to date. Due to their relatively high oxidation potential4 and ability to bind heavy metals5 and proteins,6 research efforts have been directed at understanding their roles in environment-plant interactions,7 human health,8 timber processing,9 and spirits manufacture.10

Metabolite Profiling of Soybean Seed Extracts from Near-Isogenic Low and Normal Phytate Lines Using Orthogonal Separation Strategies

Untargeted metabolomic profiling using liquid chromatography-mass spectrometry (LC-MS) was applied to lipid-depleted methanolic extracts of soybean seeds utilizing orthogonal chromatographic separations (reversed-phase and hydrophilic interaction) in both positive and negative ionization modes. Four near-isogenic lines (NILs) differing in mutations for two genes encoding highly homologous multidrug resistant proteins (MRPs) were evaluated. The double mutant exhibited a low phytate phenotype, whereas the other three NILs, the two single mutants and the wild type, did not. Principal component analysis (PCA) of the four LC-MS data sets fully separated the low phytate line from the other three. While the levels of neutral oligosaccharides were the same for all lines, there were significant metabolite differences residing in the levels of malonyl isoflavones, soyasaponins, and arginine. Two methanol-soluble polypeptides were also found as differing in abundance levels, one of which was identified as the allergen Gly m 1.