Effects of Thermobifida fusca cutinase-carbohydrate-binding module fusion proteins on cotton bioscouring

Previously, we presented a novel approach for increasing Thermobifida fusca cutinase adsorption on cotton fibers by fusing cutinase with a carbohydrate-binding module (CBM). A preliminary study showed that two fusion proteins, namely cutinase-CBM_Cel6A and cutinase-CBM_CenA, with similar stabilities and catalytic properties, had potential applications in bioscouring. In the present study, an indepth analysis of both cutinase-CBMs in bioscouring was explored. Effects of cutinase-CBMs on cotton bioscouring were investigated by characterizing the chemical and physical surface changes in enzyme-treated cotton fabrics. Gas chromatography/mass spectrometry was used to analyze the degradation of the cotton fabric cuticle; Fourier transform infrared microspectroscopy was used to study changes in the chemical composition of the cotton fabric epidermal layer; and scanning electron microscopy was used to monitor minor changes in the morphology of the fiber surface. Our results indicated that cutinase-CBMs in combination with pectinase had a greater effect on cotton fabric than did cutinase. Following scouring with cutinase-CBMs and pectinase, the performance of cotton fabric in terms of its wettability and dyeability was similar to that following alkali scouring. Our study provides a foundation for the further application of cutinase-CBM to bioscouring.     

Nanostructured biocomposite scaffolds based on collagen coelectrospun with nanohydroxyapatite

Nanofibrous biocomposite scaffolds of type I collagen and nanohydroxyapatite (nanoHA) of varying compositions (wt %) were prepared by electrostatic cospinning. The scaffolds were characterized for structure and morphology by Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), atomic force microscopy (AFM) and X-ray diffraction (XRD) techniques. The scaffolds have a porous nanofibrous morphology with random fibers in the range of 500-700 nm diameters, depending on the composition. FT-IR and XRD showed the presence of nanoHA in the fibers. The surface roughness and diameter of the fibers increased with the presence of nanoHA in biocomposite fiber as evident from AFM images. Tensile testing and nanoindendation were used for the mechanical characterization. The pure collagen fibrous matrix (without nanoHA) showed a tensile strength of 1.68 +/- 0.10 MPa and a modulus of 6.21 +/- 0.8 MPa with a strain to failure value of 55 +/- 10%. As the nanoHA content in the randomly oriented collagen nanofibers increased to 10%, the ultimate strength increased to 5 +/- 0.5 MPa and the modulus increased to 230 +/- 30 MPa. The increase in tensile modulus may be attributed to an increase in rigidity over the pure polymer when the hydroxyapatite is added and/or the resulting strong adhesion between the two materials. The vapor phase chemical crosslinking of collagens using glutaraldehyde further increased the mechanical properties as evident from nanoindentation results. A combination of nanofibrous collagen and nanohydroxyapatite that mimics the nanoscale features of the extra cellular matrix could be promising for application as scaffolds for hard tissue regeneration, especially in low or nonload bearing areas.     

Characterization of cotton fabric scouring by FT-IR ATR spectroscopy

FT-IR attenuated total reflectance (ATR) spectroscopy has been used for the fast characterization of cotton fabric scouring process. The greige and the scoured cotton fabrics showed very similar FT-IR spectrum in transmission mode because the bulk composition of the fabrics are similar. However, FT-IR ATR spectroscopy can provide information about the surface of a fabric. By examination of C–H stretching region at 2800–3000 cm⁻¹, the amount of waxes left on the fabric can be estimated. The presence of pectins and/or waxes can also be probed by observation of carbonyl peak induced by the HCl vapor treatment on the fabric. Based on these changes of FT-IR ATR spectra, the scouring process has been characterized.     

Comparative analysis of crystallinity changes in cellulose I polymers using ATR-FTIR, X-ray diffraction, and carbohydrate-binding module probes

Cotton fiber cellulose is highly crystalline and oriented; when native cellulose (cellulose I) is treated with certain alkali concentrations, intermolecular hydrogen bonds are broken and Na-cellulose I is formed. At higher alkali concentrations Na-cellulose II forms, wherein intermolecular and intramolecular hydrogen bonds are broken, ultimately resulting in cellulose II polymers. Crystallinity changes in cotton fibers were observed and assigned using attenuated total reflectance Fourier transform infrared (ATR FT-IR) spectroscopy and X-ray diffraction (XRD) subsequent to sodium hydroxide treatment and compared with an in situ protein-binding methodology using cellulose-directed carbohydrate-binding modules (CBMs). Crystallinity changes observed using CBM probes for crystalline cellulose (CBM2a, CBM3a) and amorphous cellulose (CBM4-1, CBM17) displayed close agreement with changes in crystallinity observed with ATR-FTIR techniques, but it is notable that crystallinity changes observed with CBMs are observed at lower NaOH concentrations (2.0 mol dm(-3)), indicating these probes may be more sensitive in detecting crystallinity changes than those calculated using FTIR indices. It was observed that the concentration of NaOH at which crystallinity changes occur as analyzed using the CBM labeling techniques are also lower than those observed using X-ray diffraction techniques. Analysis of crystallinity changes in cellulose using CBMs offers a new and advantageous method of qualitative and quantitative assessment of changes to the structure of cellulose that occur with sodium hydroxide treatment.     

Properties of polylactic acid composites reinforced with oil palm biomass microcrystalline cellulose

In this work, polylactic acid (PLA) composites filled with microcrystalline cellulose (MCC) from oil palm biomass were successfully prepared through solution casting. Fourier transform infrared (FT-IR) spectroscopy indicates that there are no significant changes in the peak positions, suggesting that incorporation of MCC in PLA did not result in any significant change in chemical structure of PLA. Thermogravimetric analysis was conducted on the samples. The T₅₀ decomposition temperature improved with addition of MCC, showing increase in thermal stability of the composites. The synthesized composites were characterized in terms of tensile properties. The Young's modulus increased by about 30%, while the tensile strength and elongation at break for composites decreased with addition of MCC. Scanning electron microscopy (SEM) of the composites fractured surface shows that the MCC remained as aggregates of crystalline cellulose. Atomic force microscopy (AFM) topographic image of the composite surfaces show clustering of MCC with uneven distribution.     

Conformational plasticity of the Gerstmann-Sträussler-Scheinker disease peptide as indicated by its multiple aggregation pathways

The existence of several prion strains and their capacity of overcoming species barriers seem to point to a high conformational adaptability of the prion protein. To investigate this structural plasticity, we studied here the aggregation pathways of the human prion peptide PrP82-146, a major component of the Gerstmann-Sträussler-Scheinker amyloid disease.By Fourier transform infrared (FT-IR) spectroscopy, electron microscopy, and atomic force microscopy (AFM), we monitored the time course of PrP82-146 fibril formation. After incubation at 37 °C, the unfolded peptide was found to aggregate into oligomers characterized by intermolecular β-sheet infrared bands. At a critical oligomer concentration, the emergence of a new FT-IR band allowed to detect fibril formation. A different intermolecular β-sheet interaction of the peptides in oligomers and in fibrils is, therefore, detected by FT-IR spectroscopy, which, in addition, suggests a parallel orientation of the cross β-sheet structures of PrP82-146 fibrils. By AFM, a wide distribution of PrP82-146 oligomer volumes—the smallest ones containing from 5 to 30 peptides—was observed. Interestingly, the statistical analysis of AFM data enabled us to detect a quantization in the oligomer height values differing by steps of ∼ 0.5 nm that could reflect an orientation of oligomer β-strands parallel with the sample surface. Different morphologies were also detected for fibrils that displayed high heterogeneity in their twisting periodicity and a complex hierarchical assembly.Thermal aggregation of PrP82-146 was also investigated by FT-IR spectroscopy, which indicated for these aggregates an intermolecular β-sheet interaction different from that observed for oligomers and fibrils. Unexpectedly, random aggregates, induced by solvent evaporation, were found to display a significant α-helical structure as well as several β-sheet components.All these results clearly point to a high plasticity of the PrP82-146 peptide, which was found to be capable of undergoing several aggregation pathways, with end products displaying different secondary structures and intermolecular interactions.     

Atomic force microscopy and FT-IR spectroscopy investigations of human heart valves

Human aortic, mitral, tricuspid and pulmonary heart valves were investigated by the contact mode atomic force microscopy (AFM) in air, and using FT-IR spectroscopy in the frequency range 950-4000 cm(-1). Heart valves were collected post mortem from 65-78 years old patients who died from non-cardiac diseases. All of the examined valves showed considerable heterogeneity in the surface topography of collagen fibrils as well as in their organization on the tissue surface. The AFM images revealed areas with significantly different spatial organization of the collagen fibril bundles. We observed zones with multidirectional, stacked collagen fibrils as well as areas of thin fibrils packed regularly, densely and "in phase". The majority of the collagen fibrils reproduced the typical transverse D-banding pattern, with the band interval varying in rather wide range of 70-90 nm. Using AFM imaging, objects that correspond to some pathological states of heart valves at their early stages, i.e. some forms of mineral deposits, were observed. The FT-IR spectra allowed us to recognize main components, i.e. collagen and elastin, in di.erent layers (ventricularis, fibrosa) of the valve leaflets as well as they gave also support for the presence of mineral deposits on the valve surface. The presented results showed, that the AFM imaging and FT-IR spectroscopy can be applied as a complementary methods for structural characterization of heart valves at the molecular and supramolecular levels.     

Isolation and characterization of cellulose nanofibrils from wheat straw using steam explosion coupled with high shear homogenization

Cellulose nanofibrils of diameter 10–50 nm were obtained from wheat straw using alkali steam explosion coupled with high shear homogenization. High shear results in shearing of the fiber agglomerates resulting in uniformly dispersed nanofibrils. The chemical composition of fibers at different stages were analyzed according to the ASTM standards and showed increase in α-cellulose content and decrease in lignin and hemicellulose. Structural analysis of steam exploded fibers was carried out by Fourier Transform Infrared (FT-IR) spectroscopy and X-ray diffraction (XRD). Thermal stability was higher for cellulose nanofibrils as compared to wheat straw and chemically treated fibers. The fiber diameter distribution was obtained using image analysis software. Characterization of the fibers by AFM, TEM, and SEM showed that fiber diameter decreases with treatment and final nanofibril size was 10–15 nm. FT-IR, XRD, and TGA studies confirmed the removal of hemicellulose and lignin during the chemical treatment process.     

Nanoscale structures and mechanics of barnacle cement

Polymerized barnacle glue was studied by atomic force microscopy (AFM), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and chemical staining. Nanoscale structures exhibiting rod-shaped, globular and irregularly-shaped morphologies were observed in the bulk cement of the barnacle Amphibalanus amphitrite (=Balanus amphitrite) by AFM. SEM coupled with energy dispersive X-ray (EDX) provided chemical composition information, making evident the organic nature of the rod-shaped nanoscale structures. FTIR spectroscopy gave signatures of β-sheet and random coil conformations. The mechanical properties of these nanoscale structures were also probed using force spectroscopy and indentation with AFM. Indentation data yielded higher elastic moduli for the rod-shaped structures when compared with the other structures in the bulk cement. Single molecule AFM force-extension curves on the matrix of the bulk cement often exhibited a periodic sawtooth-like profile, observed in both the extend and retract portions of the force curve. Rod-shaped structures stained with amyloid protein-selective dyes (Congo red and thioflavin-T) revealed that about 5% of the bulk cement were amyloids. A dominant 100 kDa cement protein was found to be mechanically agile, using repeating hydrophobic structures that apparently associate within the same protein or with neighbors, creating toughness on the 1–100 nm length scale.     

Changes in the structural properties and rate of hydrolysis of cotton fibers during extended enzymatic hydrolysis

An extended enzymatic hydrolysis of cotton fibers by crude cellulase from Trichoderma pseudokoningii S-38 is described with characterization of both the enzyme changes of activities and cellulose structure. The hydrolysis rates declined drastically during the early stage and then slowly and steadily throughout the whole hydrolysis process the same trend could be seen during the following re-hydrolysis process. Morphological and structural changes to the fibers, such as swelling, frequent surface erosion, and variation in the packing and orientation of microfibrils, were investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Observation of X-ray diffraction and IR spectra suggests that the hydrolysis process results in a gradual increase in the relative intensity of the hydrogen bond network, and a gradual decrease in the apparent crystal size of cellulose. The I(alpha) crystal phase was hydrolyzed more easily than was the I(beta) crystal phase. Apart from the inactivation of CBHs activity, changes in the packing and arrangement of microfibrils and the structural heterogeneity of cellulose during hydrolysis could be responsible for the reduction in the rate of reaction, especially in its later stages. The results indicate that the enzymatic hydrolysis of cellulose occurs on the outer layer of the fiber surface and that, following this, the process continues in a sub-layer manner.     

A thixotropic hydrogel from chemically cross-linked guar gum: synthesis, characterization and rheological behaviour

Polysaccharide guar gum (GG) was cross-linked in an alkaline solution with polyethylene glycol diglycidyl ether (PEGDGE) to create a new hydrogel. The GG hydrogel was examined by FT-IR spectroscopy, AFM analysis and SEM analysis. The water uptake of the GG hydrogel was measured at different pHs, and rheological studies were performed to verify the thixotropic nature of the material. Rheological studies revealed the pseudoplastic behaviour of the GG hydrogel and its thixotropic nature. AFM analysis on a sample which was subjected to shear stress showed the presence of nanoparticles in the hydrogel. When the sample was left to settle, the gel surface returned to its original homogenous morphology. The thixotropic and injectable nature of the GG hydrogel suggest its possible use in biomedical applications.     

Changes of the surface structure of corn stalk in the cooking process with active oxygen and MgO-based solid alkali as a pretreatment of its biomass conversion

This study presents a novel, efficient and environmentally friendly process for the cooking of corn stalk that uses active oxygen (O₂ and H₂O₂) and a recoverable solid alkali (MgO). The structural changes on the surface of corn stalk before and after cooking were characterized by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) techniques. The results showed that lignin and extractives were effectively removed, especially those on the surface of corn stalk. Additionally, the changes included becoming fibrillar, the exposure of cellulose and hemi-cellulose and the pitting corrosion on the surface, etc. The results also showed that the removal reaction is from outside to inside, but the main reaction is possibly on the surface. Furthermore, the results of active oxygen cooking with a solid alkali are compared with those of alkaline cooking in the paper.     

Effect of SDS on human hair: Study on the molecular structure and morphology

This paper presents a model study to understand the effect of surfactants on the physicochemical properties of human hair. FT‐IR ATR spectroscopy has been employed to understand the chemical changes induced by sodium dodecyl sulfate (SDS) on human scalp hair. In particular, the SDS induced changes in the secondary structure of protein present in the outer protective layer of hair, i.e. cuticle, have been investigated. Conformational changes in the secondary structure of protein were studied by curve fitting of the amide I band after every phase of SDS treatment. It has been found that SDS brings rearrangements in the protein backbone conformations by transforming β ‐sheet structure to random coil and β ‐turn. Additionally, AFM and SEM studies were carried out to understand the morphological changes induced on the hair surface. SEM and AFM images demonstrated the rupture and partial erosion of cuticle sublayers. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Proteolysis kinetics and structural characterization of ultrasonic pretreated sunflower protein

Proteolysis kinetics and structure attributes of sunflower meal protein (SMP) pretreated using sonication of frequency type single (20, 40 kHz) and dual (20/40 kHz) were examined. A simplified model with impeded enzyme reaction was developed and the proteolysis of SMP in a heterogeneous system was successfully described. Initial observed rate (k_in) increased after sonication, demonstrating the impacts of sonication on SMP by enhancing proteolysis (enzymolysis) and altering the structure of SMP, which was validated by circular dichroism (CD) spectroscopy, ultraviolet-visible (UV-Vis) spectroscopy, scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared (FTIR) spectroscopy and sodium dodecyl sulfide polyacrylamide gel electrophoresis (SDS-PAGE) analyses. From SMP characterization, there was a reduction in α-helix, enhancement in absorption intensity and alterations in functional groups of SMP following sonication. SEM and AFM observations indicated that sonicated SMP unfolded and exhibited more heterogenous structures, and irregular small-sized particles than control, especially at 20/40 kHz. SDS-PAGE profile displayed notable changes in molecular weight after sonication, which evidenced limited unfolding in SMP conformation. The kinetic model has possibility of being used to control enzymolysis; and structural attributes could guide in the development of ultrasonic equipment for use of pretreating protein for enzymolysis.     

Immobilization of alkaline polygalacturonate lyase from Bacillus subtilis on the surface of bacterial polyhydroxyalkanoate nano-granules

Applied Microbiology and Biotechnology - Alkaline polygalacturonate lyase (PGL), one of the pectinolytic enzymes, has been widely used for the bioscouring of cotton fibers, biodegumming, and...     

Implementation of batchwise bioscouring of cotton knits

The examination of critical factors determining the performance of bioscouring showed that a short treatment of the fabric at greater than 80°C after pectinase treatment at 60°C was essential for removal of waxes from the fabric as demonstrated by diminished intensities of methylene peaks in FT-IR measurements. Batch-wise bioscouring of cotton knits was carried out several times with post-treatment at 80°C using a rapid dyeing machine. The dye-ability of bioscoured knits was as good as the company's alkaline scoured ones with slightly higher K/S values. Water pollution caused by effluents of bioscouring and alkaline processes were estimated, as well as that due to the input of chemicals and enzymes. Higher BOD:COD_Cr ratios for enzymes indicated their biodegradable character. After calculation of energy consumption using a simulation program, an economic evaluation of the two processes was done on the basis of one ton production by considering the costs of chemicals and enzyme, water usage, energy consumption and waste water treatment charge.     

Implementation of batchwise bioscouring of cotton knits

The examination of critical factors determining the performance of bioscouring showed that a short treatment of the fabric at greater than 80°C after pectinase treatment at 60°C was essential for removal of waxes from the fabric as demonstrated by diminished intensities of methylene peaks in FT-IR measurements. Batch-wise bioscouring of cotton knits was carried out several times with post-treatment at 80°C using a rapid dyeing machine. The dye-ability of bioscoured knits was as good as the company's alkaline scoured ones with slightly higher K/S values. Water pollution caused by effluents of bioscouring and alkaline processes were estimated, as well as that due to the input of chemicals and enzymes. Higher BOD:COD_Cr ratios for enzymes indicated their biodegradable character. After calculation of energy consumption using a simulation program, an economic evaluation of the two processes was done on the basis of one ton production by considering the costs of chemicals and enzyme, water usage, energy consumption and waste water treatment charge.     

Chemical analysis of a cornstarch film surface modified by SF6 plasma treatment

SF₆ plasmas were employed to improve the water repellency of cornstarch films by producing physical as well as chemical modifications of the film surface. Samples were placed in the cathode of a capacitively coupled plasma enhanced chemical vapor deposition (PECVD) reactor. Local surface modifications resulting from SF₆ plasma treatment were evaluated using atomic force microscopy (AFM) and scanning electron microscopy (SEM). Photoelectron spectroscopy (XPS) and Raman spectroscopy were used to characterize sulfur and fluoride incorporation on the surface as well as changes in the chemical state of carbon. The results indicate that fluoride and sulfur incorporation is dependent on the self bias, and fluoride is preferentially incorporated at self-biases higher than 100 V. The carbon chemical state changed, and an amorphous-like layer was formed upon treatment. Surface reticulation was observed, indicated by the formation of a structure that resembled starch recrystallization. Optimized treatment conditions led to water contact angles over 120°.     

Degradation kinetics of pectins by an alkaline pectinase in bioscouring of cotton fabrics

Alkaline pectinases have been proven to be effective as bioscouring agents of cotton fabrics. In order to monitor the scouring degree of cotton fabrics quantificationally, a kinetic study of the degradation of pectins in cotton by an alkaline pectinase ‘Bioprep 3000L’ was performed and the influences of initial pectinase concentration and treatment time on bioscouring were evaluated quantitatively. The results showed that although the degradation products increased as pectinase concentration grew higher at same incubation time, the growth multiples of the maximum degradation rate which was used as the starting degradation rate were less than those of initial enzyme concentration. The degradation kinetics of pectins in cotton fibers with a pectinase could be described by modified Ghose–Walseth kinetic empirical equations which had been previously applied to the degradation reaction of cellulose.