15 research outputs found
Effect of Nonionic Surfactants on Dispersion and Polar Interactions in the Adsorption of Cellulases onto Lignin
Residual lignin in
pretreated biomass impedes enzymatic hydrolysis.
Nonionic surfactants are known to enhance the enzymatic hydrolysis
of lignocellulosic biomass but their mechanisms of action are incompletely
understood. This study investigates the effect of a nonionic surfactant,
Tween 80, on the adsorption of cellulases onto model lignin substrates.
Lignin substrates were prepared by spin coating of flat substrates
with three different types of lignin: organosolv lignin, kraft lignin,
and milled wood lignin. The functional group distributions in the
lignins were quantitatively analyzed by <sup>31</sup>P NMR spectroscopy.
The surface energies and surface roughnesses of the substrates were
determined by contact angle measurements and atomic force microscopy,
respectively. Tween 80 and cellulase adsorption onto the lignin substrates
was analyzed with a quartz crystal microbalance with dissipation monitoring.
Tween 80 adsorbed rapidly and primarily (≥85%) via dispersion
interactions onto the lignin substrates and effected solubilization
of lignin molecules, most notably with organosolv lignin, having the
largest dispersive surface energy component and smallest molar mass.
Cellulase adsorption onto the lignin substrates was mostly irreversible
and had both a rapid and a gradual adsorption stage. Rapid cellulase
adsorption was primarily (≥64%) mediated by dispersion interactions.
The subsequent gradual mass increase is postulated to involve swelling
of the lignin substrates. Adsorbed Tween 80 rendered lignin surfaces
more hydrophilic by increasing their polar surface energy component
and reduced both the extent of rapid cellulase adsorption as well
as the rate of the subsequent gradual mass increase. The effect of
Tween 80 on the rate and extent of the gradual mass increase depended
strongly on the chemical properties of the lignin
An Efficient, Regioselective Pathway to Cationic and Zwitterionic <i>N</i>‑Heterocyclic Cellulose Ionomers
Cationic
derivatives of cellulose and other polysaccharides are
attractive targets for biomedical applications due to their propensity
for electrostatically binding with anionic biomolecules, such as nucleic
acids and certain proteins. To date, however, relatively few practical
synthetic methods have been described for their preparation. Herein,
we report a useful and efficient strategy for cationic cellulose ester
salt preparation by the reaction of 6-bromo-6-deoxycellulose acetate
with pyridine or 1-methylimidazole. Dimethyl sulfoxide solvent favored
this displacement reaction to produce cationic cellulose acetate derivatives,
resulting in high degrees of substitution (DS) exclusively at the
C-6 position. These cationic cellulose derivatives bearing substantial,
permanent positive charge exhibit surprising thermal stability, dissolve
readily in water, and bind strongly with a hydrophilic and anionic
surface, supporting their potential for a variety of applications
such as permeation enhancement, mucoadhesion, and gene or drug delivery.
Expanding upon this chemistry, we reacted a 6-imidazolyl-6-deoxycellulose
derivative with 1,3-propane sultone to demonstrate the potential for
further elaboration to regioselectively substituted zwitterionic cellulose
derivatives
Surface-Initiated Dehydrogenative Polymerization of Monolignols: A Quartz Crystal Microbalance with Dissipation Monitoring and Atomic Force Microscopy Study
This work highlights a real-time
and label-free method to monitor
the dehydrogenative polymerization of monolignols initiated by horseradish
peroxidase (HRP) physically immobilized on surfaces using a quartz
crystal microbalance with dissipation monitoring (QCM-D). The dehydrogenative
polymer (DHP) films are expected to provide good model substrates
for studying ligninolytic enzymes. The HRP was adsorbed onto gold
or silica surfaces or onto and within porous desulfated nanocrystalline
cellulose films from an aqueous solution. Surface-immobilized HRP
retained its activity and selectivity for monolignols as coniferyl
and <i>p</i>-coumaryl alcohol underwent dehydrogenative
polymerization in the presence of hydrogen peroxide, whereas sinapyl
alcohol polymerization required the addition of a nucleophile. The
morphologies of the DHP layers on the surfaces were investigated via
atomic force microscopy (AFM). Data from QCM-D and AFM showed that
the surface-immobilized HRP-initiated dehydrogenative polymerization
of monolignols was greatly affected by the support surface, monolignol
concentration, hydrogen peroxide concentration, and temperature
Effects of Sulfate Groups on the Adsorption and Activity of Cellulases on Cellulose Substrates
Pretreatment of lignocellulosic biomass
with sulfuric acid may
leave sulfate groups on its surface that may hinder its biochemical
conversion. This study investigates the effects of sulfate groups
on cellulase adsorption onto cellulose substrates and the enzymatic
hydrolysis of these substrates. Substrates with different sulfate
group densities were prepared from H<sub>2</sub>SO<sub>4</sub>- and
HCl-hydrolyzed and partially and fully desulfated cellulose nanocrystals.
Adsorption onto and hydrolysis of the substrates was analyzed by quartz
crystal microbalance with dissipation monitoring (QCM-D). The surface
roughness of the substrates, measured by atomic force microscopy,
increased with decreasing sulfate group density, but their surface
accessibilities, measured by QCM-D H<sub>2</sub>O/D<sub>2</sub>O exchange
experiments, were similar. The adsorption of cellulose binding domains
onto sulfated substrates decreased with increasing sulfate group density,
but the adsorption of cellulases increased. The rate of hydrolysis
of sulfated substrates decreased with increasing sulfate group density.
The results indicated an inhibitory effect of sulfate groups on the
enzymatic hydrolysis of cellulose, possibly due to nonproductive binding
of the cellulases onto the substrates through electrostatic interactions
instead of their cellulose binding domains
Chitinase Activity on Amorphous Chitin Thin Films: A Quartz Crystal Microbalance with Dissipation Monitoring and Atomic Force Microscopy Study
Chitinases
are widely distributed in nature and have wide-ranging
pharmaceutical and biotechnological applications. This work highlights
a real-time and label-free method to assay Chitinase activity via
a quartz crystal microbalance with dissipation monitoring (QCM-D)
and atomic force microscopy (AFM). The chitin substrate was prepared
by spincoating a trimethylsilyl chitin solution onto a silica substrate,
followed by regeneration to amorphous chitin (RChi). The QCM-D and
AFM results clearly showed that the hydrolysis rate of RChi films
increased as Chitinase (from <i>Streptomyces griseus</i>) concentrations increased, and the optimal temperature and pH for
Chitinase activity were around 37 °C and 6–8, respectively.
The Chitinase showed greater activity on chitin substrates, having
a high degree of acetylation, than on chitosan substrates, having
a low degree of acetylation
Role of (1,3)(1,4)-β-Glucan in Cell Walls: Interaction with Cellulose
(1,3)Â(1,4)-β-d-Glucan (mixed-linkage glucan or MLG),
a characteristic hemicellulose in primary cell walls of grasses, was
investigated to determine both its role in cell walls and its interaction
with cellulose and other cell wall polysaccharides in vitro. Binding
isotherms showed that MLG adsorption onto microcrystalline cellulose
is slow, irreversible, and temperature-dependent. Measurements using
quartz crystal microbalance with dissipation monitoring showed that
MLG adsorbed irreversibly onto amorphous regenerated cellulose, forming
a thick hydrogel. Oligosaccharide profiling using <i>endo</i>-(1,3)Â(1,4)-β-glucanase indicated that there was no difference
in the frequency and distribution of (1,3) and (1,4) links in bound
and unbound MLG. The binding of MLG to cellulose was reduced if the
cellulose samples were first treated with certain cell wall polysaccharides,
such as xyloglucan and glucuronoarabinoxylan. The tethering function
of MLG in cell walls was tested by applying <i>endo</i>-(1,3)Â(1,4)-β-glucanase
to wall samples in a constant force extensometer. Cell wall extension
was not induced, which indicates that enzyme-accessible MLG does not
tether cellulose fibrils into a load-bearing network
KTN-BMP-2 interaction.
<p>Association (a) and dissociation (d) electrostatic interaction profiles of BMP-2 and the KTN monolayer in PBS and in water. BMP-2 analytes were successively flowed, at incrementally increasing concentrations: A) in PBS (pH 7.4) at 0.2, 0.4, 0.9, 1.7, 3.5, and 6.9 μM, B) in PBS (pH 4.5) at 0.03, 0.05, 0.1, 0.2, 0.4, 0.9, and 1.7 μM, and C) in water (pH 7) at 0.03, 0.05, 0.1, 0.2, and 0.4 μM. KTN-BMP-2 electrostatic attraction was strongest in water (KD = 1.1 × 10–7 M). In the presence of PBS salts at physiological pH, binding association was slightly weakened (KD = 3.2 × 10–5 M). Acidification of the PBS eliminated any binding between BMP-2 and KTN.</p
KTN bulk release.
<p>KTN gel bulk degradation in vitro at A-B) constant pH = 7.4, and C-D) constant [KCl] = 154 mM for over a period of 28 days at 37°C. Faster degradation occurred at longer time points, lower [KCl], and higher pH levels.</p
Comparison between reduced (KTN) and oxidized (KOS) keratin biomaterials.
<p>KTN can form disulfide linkages to produce a stable scaffold; but KOS cannot, due to the sulfonic acid modification of thiol groups. Consequently, the electrostatic properties are also altered, resulting in more negatively-charged KOS scaffolds, prone to more rapid hydrolytic degradation than KTN scaffolds. 10 mM NaOH solvent was used for wetting and soaking.</p
XPS analysis.
<p>A) Wide-scan XPS spectra of gold surfaces treated with 10 mM NaOH solvent, KOS, and KTN. Overnight incubation of KTN led to no detectable Au signals. The inset graph shows that, compared to the solvent group, KOS has very similar concentration levels of carbon, nitrogen, oxygen, sulfur, and gold, while KTN has elevated amounts of protein elements (C, N, and O) but decreased Au. B) Near-scan analysis displays the formation of an amide (O = C-N) peak at 288 eV, corresponding to KTN protein deposition on gold. Unbound and gold-bound KTN thiols were also detected at 163.6 and 162.5 eV, respectively. Partial adsorption of KTN on gold shifted the Au4f peaks to slightly lower energies.</p