Cellulose Ribbons Research Articles

Physical properties of bacterial cellulose aqueous suspensions treated by high pressure homogenizer

In the present study, purified wet bacterial cellulose was disrupted into cellulose aqueous suspensions (0.5%, w/v) and then homogenized by 10 passes at 0, 200, 400 and 600 bar respectively. In order to evaluate the effects of high pressure homogenization (HPH) on bacterial cellulose aqueous suspensions, morphology, rheology, stability, texture, water holding capacity (WHC), water swelling ability (WSA) and water release rate (WRR) were investigated. Morphological analysis by atomic force microscopy revealed changes in microstructure and dispersion of cellulose ribbons after HPH treatment. The diameter of micro-fibril ribbons decreased from 95.6 nm to 60.3 nm with increasing pressure up to 600 bar. The rheology results suggested that all the suspensions displayed a shear thinning behaviour, moreover the suspensions treated by HPH showed 3 regions (shear thinning region, plateau region and shear thinning region again) and the flow curves followed the Herschel–Bulkley model. Furthermore, the stability and textural properties of the suspensions were enhanced after HPH treatment. Additionally, WHC and WSA of cellulose suspensions displayed high stability and the WRR increased after HPH treatment during storage.

Characterization of water-soluble exopolysaccharides from Gluconacetobacter xylinus and their impacts on bacterial cellulose crystallization and ribbon assembly

Gluconacetobacterxylinus has the ability to produce different types of water soluble exopolysaccharides (EPS). Those EPS have different levels of association to bacterial cellulose (BC). At least a portion of the EPS can be released from the BC by 0.1 to 4 M NaOH solution treatments. Hard to extract EPS (HE-EPS) released by 4 M NaOH solutions have been characterized and contain approximately 75 % mannose and 25 % glucose. To study the effect of the EPS on BC synthesis, purified EPS were added to the medium at the start of cultivation and the BC produced was characterized. Results showed that the presence of HE-EPS in the culture medium interfered with the alignment of the BC crystals, but did not reduce crystal size. This is in contrast to similar studies performed using xyloglucan, xylan and glucomannan. The width of the average ribbon increased by 60 % when HE-EPS levels increased in the medium, which indicated that the HE-EPS could also modulate the bundling of cellulose ribbons. Based on the data we propose a mechanism for how HE-EPS alters cellulose formation and assembly. The addition of HE-EPS disturbs the preferential crystal orientation and increases the spacing of cellulose microfibrils without affecting crystallization by associating with ordered cellulose prior to physical aggregation or bundling.

Bacterial cellulose/gelatin composites: in situ preparation and glutaraldehyde treatment

Bacterial cellulose (BC)/GEL composites were prepared in situ by adding gelatin into BC-producing culture medium. The addition of gelatin interfered with the formation of the BC pellicle structure and thus made the BC yield and growth rate quite different from that of pure BC. Scanning electron microscope images showed that the width of cellulose ribbons became narrower than that of pure BC and the gelatin filled in the pores of BC to form a dense structure. The addition level of gelatin significantly influences the yield of BC/GEL composites. An optimum value of 0.5 wt/v% gelatin was attained, with which the highest yield of 0.0541 g/100 mL was achieved. Under this condition, the weight percentage of gelatin in BC/GEL composite was 65 wt%. BC/GEL composites were treated with glutaraldehyde to crosslink BC fibrils and gelatin. The crosslinking degree, determined by the concentration of glutaraldehyde and crosslinking time, could affect the swelling behavior, thermal stability and mechanical properties of composites. With increasing of the crosslinking degree, the crystallinity index and swelling behavior of the composites decreased. The increase in the crosslinking degree also descreased the composite’s strain at break in elongation but increased the compressive and tensile strength. Covalent bonding between BC and gelatin provides good strength retention to the glutaraldehyde-treated composites with a high crosslinking degree. Considering the cytocompatibility and properties of composites, the most appropriate concentration of glutaraldehyde and crosslinking time were 1.0 wt/v% and 24 h, respectively.

Population Balance Model Validation and Predictionof CQAs for Continuous Milling Processes: toward QbDin Pharmaceutical Drug Product Manufacturing

Continuous tablet manufacturing has been investigated for its potential advantages (e.g., cost, efficiency, and controllability) over more conventional batch processes. One avenue for tablet manufacturing involves roller compaction followed by milling to form compactible granules. A better understanding of these powder processes is needed to implement Quality by Design in pharmaceutical manufacturing. In this study, ribbons of microcrystalline cellulose were produced by roller compaction and milled in a conical screen mill. A full factorial experiment was performed to evaluate the effects of ribbon density, screen size, and impeller speed on the product size distribution and steady-state mass holdup of the mill. A population balance model was developed to simulate the milling process, and a parameter estimation technique was used to calibrate the model with a subset of experimental data. The calibrated model was then simulated at other processing conditions and compared with additional unused experimental data. Statistical analyses of the results showed good agreement, demonstrating the model’s predictive capability in quantifying milled product critical quality attributes within the experimental design space. This approach can be used to optimize the design space of the process, enabling Quality by Design.

Microstructure and dielectric properties of biocarbon nanofiber composites

A kind of web-like carbon with interconnected nanoribbons was fabricated using bacterial cellulose pyrolyzed at various temperatures, and the microwave dielectric properties were investigated. Bacterial cellulose was converted into carbonized bacterial cellulose (CBC) with a novel three-dimensional web built of entangled and interconnected cellulose ribbons when the carbonization temperature was below 1,200°C; the web-like structure was destroyed at a temperature of 1,400°C. Composites of CBC impregnated with paraffin wax exhibited high complex permittivity over a frequency range of 2 to 18 GHz, depending on the carbonization temperature. Both real and imaginary parts were the highest for CBC pyrolyzed at 1,200°C. The complex permittivity also strongly depended on CBC loadings. For 7.5 wt.% loading, the real and imaginary permittivities were about 12 and 4.3, respectively, and the minimum reflection loss was -39 dB at 10.9 GHz. For 30 wt.% loading, the real and imaginary permittivities were about 45 and 80, respectively, and the shielding efficiency was more than 24 dB in the measured frequency range and could be up to 39 dB at 18 GHz. The electromagnetic properties were assumed to correlate with both the dielectric relaxation and the novel web-like structure.

Evaluation of Three Approaches for Real-Time Monitoring of Roller Compaction with Near-Infrared Spectroscopy

Three different approaches have been evaluated for monitoring ribbon density through real-time near-infrared spectroscopy measurements. The roll compactor was operated to produce microcrystalline cellulose (MCC) ribbons of varying densities. The first approach used the slope of the spectra which showed a variation through the ribbon that could be attributed to density. A second qualitative approach was also developed with a principal component analysis (PCA) model with spectra taken in-line during the production of ribbons in an ideal roll pressure range. The PCA (i.e., real-time) density scans show that the model was able to qualitatively capture the density responses resulting from variation in process parameters. The third approach involved multivariate partial least squares (PLS) calibration models developed at wavelength regions of 1,120-1,310 and 1,305-2,205 nm. Also, various PLS models were developed using three reference methods: caliper, pycnometer, and in-line laser. The third approach shows a quantitative difference between the model-predicted and the measured densities. Models developed at high-wavelength region showed highest accuracy compared with models at low-wavelength region. All the PLS models showed a high accuracy along the spectra collected throughout the production of the ribbons. The three methods showed applicability to process control monitoring by describing the changes in density during in-line sampling.

Electromagnetic performances of composites with promising carbons derived from bacterial cellulose

Carbon-matrix composites with interconnected carbon nanoribbon filler were fabricated using bacterial cellulose as the carbon precursor, and their use for electromagnetic interference shielding and microwave absorption was investigated. After carbonization at 800°C, bacterial cellulose (BC) was converted into a carbon (CBC) with a novel three-dimensional web built of entangled and interconnected cellulose ribbons. Composites of CBC impregnated with paraffin wax exhibit high complex permittivity over a frequency range of 2–18GHz, depending on the CBC content. The shielding effectiveness reaches 20dB in the measured frequency range for the 40wt.% CBC/paraffin wax composite with only a 1mm thickness. Rhombic shape Fe3O4 nanoparticles were prepared by oxidation precipitation and mixed with paraffin wax in a mass ratio of 2:1. Incorporating 2.5wt.% CBC into the Fe3O4/paraffin wax composite decreases the reflection loss at least 5 times. CBC dramatically improves the dielectric loss and attenuation constant through the increase of imaginary permittivity, leading to improved microwave-absorption properties.

Assessment of the critical factors affecting the porosity of roller compacted ribbons and the feasibility of using NIR chemical imaging to evaluate the porosity distribution

The purpose of this study was to assess the porosity variation of roller compacted ribbons made using different process parameters; in addition, the feasibility of using near-infrared chemical imaging (NIR-CI) to evaluate porosity variations was examined. Ribbons of neat microcrystalline cellulose were compacted using a range of roll pressures (RP), roll speeds (RS) and feed screw speeds (FSS). The ribbon porosity decreased as RP increased with the exception of ribbons produced by the combination of high RS and low FSS where increasing RP increases the porosity of the ribbons. Lower RS was found to produce ribbons with lower porosity and the porosity increases as the RS increased. Increased FSS will decrease ribbon porosity at higher RS while it slightly increase the ribbon porosity at lower RS. A simple linear regression model showed NIR-CI was able to predict the ribbon porosity with a correlation of 0.9258. NIR-CI is able to characterize differences in porosity as a function of position on the ribbon where regions with lower porosity show higher absorbance. Nevertheless, NIR-CI is able to show sinusoidal variation in intensities along the roller compacted ribbon among all settings studied.

Interaction between nanosized crystalline components of a composite based on Acetobacter xylinum cellulose and calcium phosphates

A composite consisting of two nanosized biocompatible components, Acetobacter xylinum cellulose and calcium phosphate, is prepared through aggregation in an aqueous suspension. The structures of initial components and composite are investigated by the methods of X-ray and electron diffraction and electron microscopy. The mineral component consists of two crystalline phases, hydroxyapatite and whitlockite (magnesium-containing tricalcium phosphate), which are nanosized platelike crystals. The composite preserves the crystalline structures of initial calcium phosphates and cellulose. In the course of composite formation, hydroxyapatite and whitlockite crystallites are adsorbed on the surfaces of nanofibrillar cellulose ribbons. Whitlockite nanocrystals are predominantly deposited on the surface of cellulose ribbons. The mutual orientation of the surfaces of crystalline structures of cellulose and two types of calcium phosphates, hydroxyapatite and whitlockite, is analyzed by means of computer simulation, and the variants of mutual arrangement of their surfaces during formation of the interfacial boundary are suggested.

Adding enzymatically modified gelatin to enhance the rehydration abilities and mechanical properties of bacterial cellulose

Bacterial cellulose (BC) possesses excellent water holding capacity. However, increased crystallization causes a decrease in the rehydration ability of dried BC. Due to its excellent hydrophilic properties, we used gelatin and its enzymatically modified form (EMG) to prepare BC nano-composites in an attempt to enhance the rehydration abilities properties of BC. The polar peptide fraction was increased by extended hydrolysis of fish gelatin in Alcalase. Peptides smaller than 10 kDa were obtained by hydrolysis at 50 °C for 20 min. Protein contents of composites prepared by immersing BC in 5% gelatin (Gelatin/BC) or EMG (EMG/BC) solution were 81% and 92%, respectively, both of which then formed high gelatin/BC composites (HGBC). The protein content of EMG/BC was higher than that of Gelatin/BC, as compared with low gelatin/BC composites (LGBC) that were formed by immersion in a corresponding 0.5% solution. Among both HGBC and LGBC composites, EMG/BC exhibited the best rehydration abilities. Freeze-dried EMG/BC also exhibited the fastest rehydration rate and the best ability to restore wet-type composites. Composite microstructures revealed that BC was enveloped by gelatin when non-polar EMG (NPEMG) and polar EMG (PEMG) entered the BC network and adsorbed onto cellulose ribbons. The microstructure of EMG/BC contained both PEMG/BC and NPEMG/BC structures. Gelatin hydrolysates, penetrating BC networks and forming stable composites, improved the rehydration ability of dried BC. The polar functional groups of gelatin and its hydrolysates represent the key factors contributing to the hydrophilic nature of composites.

Characterisation of density distributions in roller-compacted ribbons using micro-indentation and X-ray micro-computed tomography

Roller compaction is one stage in a dry granulation process to produce free flowing granules. Its proper understanding is essential in optimising manufacturing efficiency and product quality. Roller compaction produces a compacted strip or “ribbon”, which is then milled to produce granules. For a given milling condition, the density distribution in the ribbons determines the properties of the granules (particularly their size distribution and strength). Therefore, knowing the density distributions in the ribbons is very important in improving the effectiveness of the roller compaction process and the quality of the granules produced. In this paper, the density distribution in roller-compacted ribbons of microcrystalline cellulose (Avicel PH102) has been examined using three different techniques: (1) sectioning; (2) micro-indentation and (3) X-ray micro-computed tomography. It has been shown that with proper calibration all three techniques can essentially produce the same results, but with a different degree of resolution (scale of scrutiny). In addition, the influence of process conditions, such as roll gap, roll speed and the presence or absence of lubrication, on the ribbon density distributions has also been investigated. Flow into the press is often constrained by the presence of “cheek plates”, which prevent lateral powder movement. In this type of arrangement, it is found that non-uniform powder feeding occurs in the compaction region, induced by the friction between the powder and the cheek plates; as a result, the densities in the middle of the ribbon width are generally higher than those close to the edges. It has also been shown that higher average ribbon densities are obtained when the roll gap, roll speed, or the friction between the powder and the side cheek plates is reduced.

Bacterium organizes hierarchical amorphous structure in microbial cellulose

A pellicle, a gel film of microbial cellulose, is a supermolecular system containing 99% of water by weight, which is closely related to an amorphous structure in it. Using ultra-small-angle neutron scattering, in order to cover over a wide range of length scales from nm to 10 microm, we examined the hierarchical amorphous structure in the microbial cellulose, which is synthesized by a bacterium (Acetobacter xylinum). The microbial cellulose swollen by water shows small-angle scattering that obeys a power law q -behavior according to q -alpha as a function of the magnitude of the scattering vector q . The power law, determined by scattering, is attributed to a mass fractal due to the distribution of the center of mass for the crystallite (microfibril) in amorphous cellulose swollen by water. As q increases, alpha takes the values of 2.5, 1, and 2.35, corresponding, respectively, to a gel network composed of bundles, a bundle composed of cellulose ribbons, and concentration fluctuations in a bundle. From the mass fractal q -behavior and its length scale limits, we evaluated a volume fraction of crystallite in microbial cellulose. It was found that 90% of the cellulose bundle is occupied by amorphous cellulose containing water.

Cellulose synthesized by Acetobacter xylinum in the presence of multi-walled carbon nanotubes

The structure of bacterial cellulose is affected by the bacterial strain used, culture media and cultivation conditions. In this study, acid-treated multi-walled carbon nanotubes (MWNTs) were added into a static culture medium and their effect on bacterial cellulose structure was studied by scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FT-IR), CP/MAS 13C NMR and X-ray diffractometry. The bacterial cellulose ribbons and the MWNTs interwound and formed a three-dimensional network architecture. Band-like assemblies with sharp bends and rigidity were also produced in the presence of MWNTs. The intermolecular hydrogen bonds in bacterial cellulose produced in the presence of MWNTs were weakened. The crystal structure, cellulose I α content, crystallinity index (CrI) and crystallite size all changed. The results may suggest that the acid-treated MWNTs containing hydroxyl groups interact with the sub-elementary bacterial cellulose fibrils, subsequently interfering with the aggregation and crystallization.

Application of thermal effusivity as a process analytical technology tool for monitoring and control of the roller compaction process

The aim of this study was to examine the relationship between physical characteristics of compacted ribbons and their thermal effusivity in an attempt to evaluate the feasibility of using effusivity for in-process monitoring of roller compaction. In this study, thermal effusivity, solid fraction, tensile strength, and Young's modulus of ribbons of microcrystalline cellulose (MCC), anhydrous lactose, and placebo (PBO) formulations containing various ratios of MCC to anhydrous lactose (75:20, 55:40, 40:55, and 20:75) were determined at various compaction pressures (25-150 bars). The effusivity-square root of solid fraction relationship was linear for MCC and all the PBO formulations but was a second-order polynomial function for lactose. This could be due to the predominant deformation of lactose by brittle fracture, which might have significantly increased the number and size of contact points between particles, causing a change in thermal conductivity along with a density change. The effusivity-tensile strength and effusivity-Young's modulus relationships were best described by logarithmic functions for MCC but were linear for lactose up to a compaction pressure of 65 bars. There were similar relationships for effusivity with tensile strength and Young's modulus for all PBO formulations except PBO IV, which might have been due to the deformation of lactose, the largest component in this formulation. Strong correlations between effusivity and physical properties of ribbons were established. Although these correlations were formulation-dependent, they demonstrate the possibility of using effusivity as a tool in monitoring roller compaction.

Features of bacterial cellulose synthesis in a mutant generated by disruption of the diguanylate cyclase 1 gene of Acetobacter xylinum BPR 2001.

The diguanylate cyclase 1 (DGC1) (dgc1) gene in Acetobacter xylinum BPR 2001--a bacterial cellulose (BC) producer--was cloned and sequenced, and a DGC1 gene-disrupted mutant, strain DD, was constructed. The production and structural characteristics of the BC formed by DD were compared with those of the parental strain BPR 2001. BC production by DD was almost the same as that by BPR 2001 in static cultivation and in shake flask cultivation. However, in a jar fermentor DD produced about 36% more BC than the parental strain. DD produced suspended particle materials that cannot aggregate owing to their random structural characteristics in static cultivation; more uniformly dispersed BC pellicles and smaller BC pellets are produced on average in a jar fermentor, as reflected by the higher BC production by DD than by the parental strain in a jar fermentor. Micrographs of BC produced by DD revealed that the width of cellulose ribbons assemblies decreased as a result of differences in the ultrastructure and mechanism of formation of BC between the two strains. These results reveal that disruption of the dgc1 gene, which catalyzes synthesis of c-di-GMP (an effector of BC synthase), is not fatal for BC synthesis, although it affects BC structure.

Effects of endogenous endo-β-1,4-glucanase on cellulose biosynthesis in Acetobacter xylinum ATCC23769

Endo-β-1,4-glucanase (CMCax; EC 3.2.1.4) from Acetobacter xylinum ATCC23769 was expressed as a 6 × His-tagged fusion protein in Escherichia coli. The optimal temperature, pH, K m and V max of the purified His-tagged CMCax toward carboxymethyl cellulose were 50°C, 4.5, 20 mg/ml and 37.2 μM/min, respectively. The number of recognition residues of cello-oligosaccharide by this enzyme were five (cellopentaose) or longer, and the stereochemical course of hydrolysis was of the inverting type. Addition of a small amount (1.5 mg/l) of His-tagged CMCax into a culture medium enhanced cellulose production 1.2-fold. CMCax overproduction in A. xylinum also enhanced the yield of cellulose production. Transmission electron microscopic analysis revealed that the cellulose ribbons secreted from the CMCax overproducing strain were dispersed compared with those from the wild type strain in the same manner as by carboxymethyl cellulose addition. These results could suggest that CMCax from A. xylinum influences in cellulose ribbon assembly, which is considered to be a rate-determined process in cellulose synthesis.

Effects of Endogenous Endo-β-1,4-Glucanase on Cellulose Biosynthesis in Acetobacter xylinum ATCC23769

Endo-beta-1,4-glucanase (CMCax; EC 3.2.1.4) from Acetobacter xylinum ATCC23769 was expressed as a 6 x His-tagged fusion protein in Escherichia coli. The optimal temperature, pH, K(m) and V(max) of the purified His-tagged CMCax toward carboxymethyl cellulose were 50 degrees C, 4.5, 20 mg/ml and 37.2 microM/min, respectively. The number of recognition residues of cello-oligosaccharide by this enzyme were five (cellopentaose) or longer, and the stereochemical course of hydrolysis was of the inverting type. Addition of a small amount (1.5 mg/l) of His-tagged CMCax into a culture medium enhanced cellulose production 1.2-fold. CMCax overproduction in A. xylinum also enhanced the yield of cellulose production. Transmission electron microscopic analysis revealed that the cellulose ribbons secreted from the CMCax overproducing strain were dispersed compared with those from the wild type strain in the same manner as by carboxymethyl cellulose addition. These results could suggest that CMCax from A. xylinum influences in cellulose ribbon assembly, which is considered to be a rate-determined process in cellulose synthesis.

Cellulose synthesized by Acetobacter xylinum in the presence of plant cell wall polysaccharides

Acetobacter xylinum was cultured in Schramm–Hestrin (SH) medium containing glucuronoxylan (xylan medium) or pectin (pectin medium). Loose bundles of cellulose microfibrils were formed in the xylan medium. In contrast the cellulose ribbons formed in the pectin medium were the same as those normally formed in SH medium.The periodic acidthiocarbohydrazidesilver proteinate method indicated that positive reacted substances located along cellulose microfibrils formed in both mediums. Freeze-fracture and deepetching electron microscopy also revealed that polysaccharides exist around cellulose microfibrils. X-ray diffractometry and Fourier Transform In-frared spectroscopy demonstrated that the addition of xylan induced a change in the ratio of cellulose Iα and Iβ. Electron diffraction analysis revealed that xylan discontinuously affected the crystalline structure of cellulose microfibrils. Pectin did not have this effect. Glucuronoxylan in the medium prevented the assembly of cellulose

Rotation of cellulose ribbons during degradation with fungal cellulase

Degradation of bacterial cellulose with a commercial cellulase, Celluclast 1.5L (Novo Nordisk), from the fungus Trichoderma reesei, causes a rotational movement of the cellulose microfibrils. Purified cellulases (CBH I, CBH II, and EG II) do not induce rotation of bacterial cellulose, however, ratios of CBH I and EG II do cause rotation of bacterial cellulose. Equimolar amounts of CBH I or CBH II and EG II do not result in motion during degradation. Based on these observations, we provide further evidence supporting, at least on theoretical grounds, the hypothesis that cellulose chains have intrinsic chirality. As the cellulase enzymes interact with and degrade the cellulose fibrils, the crystalline structure of the cellulose is altered, allowing the linear cellulose polymers to relax into a lower energy state, thus relieving the strain induced by crystallization of the nascent β-glucan chains during the biogenesis of the microfibril. This conversion of crystalline bacterial ribbons into more relaxed conformations produces the rotation observed during the treatment of bacterial cellulose with cellulase.

Optimized mixtures of recombinant Humicola insolens cellulases for the biodegradation of crystalline cellulose.

The digestion of bacterial cellulose ribbons by ternary mixtures of enzymes consisting of recombinant cellulases (two cellobiohydrolases, Cel6A and Cel7A, and the endoglucanase Cel45A) from Humicola insolens was investigated over a wide range of mixture composition. The extent of digestion was followed by soluble sugar release (saccharification) analysis together with transmission electron microscopy (TEM) observations. It was found that the addition of minute quantities of Cel45A induced a spectacular increase in saccharification of the substrate with either Cel7A or the mixture of Cel6A and Cel7A. Conversely, only a moderate saccharification resulted from the mixing of Cel45A and Cel6A. This difference is believed to originate from (1) the occasional endo character of Cel6A and (2) the competition of Cel6A and Cel45A for the substrate sites that are sensitive to endo activity. Interestingly, the mixture of enzymes giving rise to the highest saccharification rate did not always correspond to mixtures of enzymes generating the highest synergy. TEM images revealed that the bacterial cellulose ribbons became at the same time cut and narrowed down under the action of an optimized mixture of the three enzymes.