Metabolic engineering of p-hydroxybenzoate in poplar lignin.

Ester-linked p-hydroxybenzoate occurs naturally in poplar lignin as pendent groups that can be released by mild alkaline hydrolysis. These 'clip-off' phenolics can be separated from biomass and upgraded into diverse high-value bioproducts. We introduced a bacterial chorismate pyruvate lyase gene into transgenic poplar trees with the aim of producing more p-hydroxybenzoate from chorismate, itself a metabolic precursor to lignin.

Direct analysis by time-of-flight secondary ion mass spectrometry reveals action of bacterial laccase-mediator systems on both hardwood and softwood samples.

The modification and degradation of lignin play a vital role in carbon cycling as well as production of biofuels and bioproducts. The possibility of using bacterial laccases for the oxidation of lignin offers a route to utilize existing industrial protein expression techniques. However, bacterial laccases are most frequently studied on small model compounds that do not capture the complexity of lignocellulosic materials.

Challenges Determining the Correct Deposition Order of Different Intersecting Black Inks by Time-of-Flight Secondary Ion Mass Spectrometry.

The distinction of different inks and determination of their deposition order are important forensic tasks when evaluating questioned documents. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) has received attention for these tasks due to the technique's nondestructive nature, rich mass spectral information, ability to provide chemical images, and excellent surface sensitivity. Prior literature results demonstrate the ability of ToF-SIMS to differentiate between many varieties of blue ballpoint pens and to determine the correct deposition order for selected ink intersections.

Comparative analysis of lignin peroxidase and manganese peroxidase activity on coniferous and deciduous wood using ToF-SIMS.

White-rot fungi are distinguished by their ability to efficiently degrade lignin via lignin-modifying type II peroxidases, including manganese peroxidase (MnP) and lignin peroxidase (LiP). In the present study, time-of flight secondary ion mass spectrometry (ToF-SIMS) was used to evaluate lignin modification in three coniferous and three deciduous wood preparations following treatment with commercial preparations of LiP and MnP from two different white-rot fungi. Percent modification of lignin was calculated as a loss of intact methoxylated lignin over nonfunctionalized aromatic rings, which is consistent with oxidative cleavage of methoxy moieties within the lignin structure.

Direct and up-close views of plant cell walls show a leading role for lignin-modifying enzymes on ensuing xylanases.

Background: A key barrier that limits the full potential of biological processes to create new, sustainable materials and fuels from plant fibre is limited enzyme accessibility to polysaccharides and lignin that characterize lignocellulose networks. Moreover, the heterogeneity of lignocellulosic substrates means that different enzyme combinations might be required for efficient transformation of different plant resources. Analytical techniques with high chemical sensitivity and spatial resolution that permit direct characterization of solid samples could help overcome these challenges by allowing direct visualization of enzyme action within plant fibre, thereby identify barriers to enzyme action.

Advancing lignocellulose bioconversion through direct assessment of enzyme action on insoluble substrates.

Microbial utilization of lignocellulose from plant cell walls is integral to carbon cycling on Earth. Correspondingly, secreted enzymes that initiate lignocellulose depolymerization serve a crucial step in the bioconversion of lignocellulosic biomass to fuels and chemicals. Genome and metagenome sequencing efforts that span the past decade reveal the diversity of enzymes that have evolved to transform lignocellulose from wood, herbaceous plants and grasses.

Towards practical time-of-flight secondary ion mass spectrometry lignocellulolytic enzyme assays.

Background: Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) is a surface sensitive mass spectrometry technique with potential strengths as a method for detecting enzymatic activity on solid materials. In particular, ToF-SIMS has been applied to detect the enzymatic degradation of woody lignocellulose. Proof-of-principle experiments previously demonstrated the detection of both lignin-degrading and cellulose-degrading enzymes on solvent-extracted hardwood and softwood.

Application of time-of-flight-secondary ion mass spectrometry for the detection of enzyme activity on solid wood substrates.

Time-of-flight-secondary ion mass spectrometry (TOF-SIMS) is a surface analysis technique that is herein demonstrated to be a viable tool for the detection of enzyme activity on solid substrates. Proof-of-principle experiments are presented that utilize commercial cellulase and laccase enzymes, which are known to modify major polymeric components of wood (i.e.

Mode of coniferous wood decay by the white rot fungus Phanerochaete carnosa as elucidated by FTIR and ToF-SIMS.

The softwood degrading white-rot fungus, Phanerochaete carnosa, was investigated for its ability to degrade two coniferous woods: balsam fir and lodgepole pine. P. carnosa grew similarly on these wood species, and like the hardwood-degrading white-rot fungus Ceriporiopsis subvermispora, P.

Expanding the library of secondary ions that distinguish lignin and polysaccharides in time-of-flight secondary ion mass spectrometry analysis of wood.

Extracted pine (Pinus spp.) wood and the holocellulose and cellulose fractions of pine were analyzed by time-of-flight secondary ion mass spectrometry (ToF-SIMS). The main sources of variation among wood constituents were elucidated by principal component analysis (PCA).

Formation of high-mass cluster ions from compound semiconductors using time-of-flight secondary ion mass spectrometry with cluster primary ions.

The detection of high-mass, nonstoichiometric, GaxAsy and InxPy secondary ion clusters using time-of-flight secondary ion mass spectrometry is reported for the first time. The GaxAsy and InxPy clusters are detected in both positive and negative ion spectra and extend to masses of at least 6000 dalton (Da). Consecutive clusters differ by the addition of one gallium (indium) atom.