crystalline cellulose
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Polymers ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 16
Author(s):  
Jesus Valcarcel ◽  
José Antonio Vázquez ◽  
Uxía R. Varela ◽  
Rui L. Reis ◽  
Ramon Novoa-Carballal

Styela clava is an edible sea squirt farmed in Korea that has gradually invaded other seas, negatively impacting the ecology and economy of coastal areas. Extracts from S. clava have shown wide bioactivities, and ascidians have the unique capability among animals of biosynthesizing cellulose. Thus, S. clava is a relevant candidate for valorization. Herein, we aimed at surveying and characterizing polysaccharides in both tunic and flesh of this ascidian. To this end, we enzymatically hydrolyzed both tissues, recovering crystalline cellulose from the tunic with high aspect ratios, based on results from microscopy, X-ray diffraction, and infrared spectroscopy analyses. Alkaline hydroalcoholic precipitation was applied to isolate the polysaccharide fraction that was characterized by gel permeation chromatography (with light scattering detection) and NMR. These techniques allowed the identification of glycogen in the flesh with an estimated Mw of 7 MDa. Tunic polysaccharides consisted of two fractions of different Mw. Application of Diffusion-Ordered NMR allowed spectroscopically separating the low-molecular-weight fraction to analyze the major component of an estimated Mw of 40–66 kDa. We identified six different sugar residues, although its complexity prevented the determination of the complete structure and connectivities of the residues. The two more abundant residues were N-acetylated and possibly components of the glycosaminoglycan-like (GAG-like) family, showing the remaining similarities to sulfated galactans. Therefore, Styela clava appears as a source of nanocrystalline cellulose and GAG-like polysaccharides.


2021 ◽  
Vol 12 ◽  
Author(s):  
Yunjun Zhao ◽  
Xiao-Hong Yu ◽  
Chang-Jun Liu

Lignin in Populus species is acylated with p-hydroxybenzoate. Monolignol p-hydroxybenzoyltransferase 1 (PHBMT1) mediates p-hydroxybenzoylation of sinapyl alcohol, eventually leading to the modification of syringyl lignin subunits. Angiosperm trees upon gravistimulation undergo the re-orientation of their growth along with the production of specialized secondary xylem, i.e., tension wood (TW), that generates tensile force to pull the inclined stem or leaning branch upward. Sporadic evidence suggests that angiosperm TW contains relatively a high percentage of syringyl lignin and lignin-bound p-hydroxybenzoate. However, whether such lignin modification plays a role in gravitropic response remains unclear. By imposing mechanical bending and/or gravitropic stimuli to the hybrid aspens in the wild type (WT), lignin p-hydroxybenzoate deficient, and p-hydroxybenzoate overproduction plants, we examined the responses of plants to gravitropic/mechanical stress and their cell wall composition changes. We revealed that mechanical bending or gravitropic stimulation not only induced the overproduction of crystalline cellulose fibers and increased the relative abundance of syringyl lignin, but also significantly induced the expression of PHBMT1 and the increased accumulation of p-hydroxybenzoates in TW. Furthermore, we found that although disturbing lignin-bound p-hydroxybenzoate accumulation in the PHBMT1 knockout and overexpression (OE) poplars did not affect the major chemical composition shifts of the cell walls in their TW as occurred in the WT plants, depletion of p-hydroxybenzoates intensified the gravitropic curving of the plantlets in response to gravistimulation, evident with the enhanced stem secant bending angle. By contrast, hyperaccumulation of p-hydroxybenzoates mitigated gravitropic response. These data suggest that PHBMT1-mediated lignin modification is involved in the regulation of poplar gravitropic response and, likely by compromising gravitropism and/or enhancing autotropism, negatively coordinates the action of TW cellulose fibers to control the poplar wood deformation and plant growth.


Holzforschung ◽  
2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Yuko Ono ◽  
Miyuki Takeuchi ◽  
Yaxin Zhou ◽  
Akira Isogai

Abstract Eucalyptus (Eucalyptus globulus) cellulose was isolated from wood powder by dewaxing, delignification, and subsequent 4% NaOH extraction. 2,2,6,6-Tetramethyl-piperidine-1-oxyl (TEMPO)-oxidized eucalyptus celluloses were prepared from never-dried eucalyptus cellulose (EC) in yields of 96% and 72% (based on the dry weight of EC) when oxidized with NaOCl of 5 and 10 mmol/g-EC, respectively. Their carboxy contents were 1.4 and 1.8 mmol/g, respectively, when determined by conductivity titration. The crystallinity of cellulose I for EC decreased by TEMPO-mediated oxidation, showing that the originally crystalline region in EC was partly converted to disordered regions by TEMPO-mediated oxidation. Correspondingly, the relative signal area of C6‒OH/C1 with the trans-gauche (tg) conformation attributed to crystalline cellulose I in the solid-state 13C NMR spectrum of EC decreased from 0.42 to 0.34 by TEMPO-mediated oxidation with NaOCl of 10 mmol/g-EC. TEMPO-oxidized EC prepared with NaOCl of 10 mmol/g-EC was almost completely converted into individual TEMPO-oxidized EC nanofibrils (TEMPO-ECNFs) of homogeneous widths of ∼3 nm widths and lengths of >1 μm by mechanical disintegration in water. However, the TEMPO-ECNFs contained many kinks and had uneven surfaces, probably owing to significant damage occurring on the surface cellulose molecules of crystalline cellulose microfibrils during TEMPO-mediated oxidation.


2021 ◽  
Author(s):  
Chao Zhong ◽  
Bernd Nidetzky

AbstractEnzyme-catalyzed iterative β-1,4-glycosylation of β-glycosides is promising for bottom-up polymerization of reducing-end-modified cello-oligosaccharide chains. Self-assembly of the chains from solution yields crystalline nanocellulose materials with properties that are tunable by the glycoside group used. Cellulose chains with a reducing-end thiol group are of interest to install a controllable pattern of site-selective modifications into the nanocellulose material. Selection of the polymerizing enzyme (cellodextrin phosphorylase; CdP) was pursued here to enhance the synthetic precision of β-1-thio-glucose conversion to generate pure “1-thio-cellulose” (≥95%) unencumbered by plain (unlabeled) cellulose resulting from enzymatic side reactions. The CdP from Clostridium stercorarium (CsCdP) was 21 times more active on β-1-thio-glucose (0.17 U/mg; 45 °C) than the CdP from Clostridium cellulosi (CcCdP), and it lacked hydrolase activity, which is substantial in CcCdP, against the α-d-glucose 1-phosphate donor substrate. The combination of these enzyme properties indicated that CsCdP is a practical catalyst for 1-thio-cellulose synthesis directly from β-1-thio-glucose (8 h; 25 mol% yield) that does not require a second enzyme (cellobiose phosphorylase), which was essential when using the less selective CcCdP. The 1-thio-cellulose chains had an average degree of polymerization of ∼10 and were assembled into highly crystalline cellulose II crystallinity material.


2021 ◽  
Vol 10 (15) ◽  
pp. e526101523267
Author(s):  
Letícia Pereira dos Santos Barbosa de Sousa ◽  
Priscila Maria Sarmeiro Correa Marciano Leite ◽  
Angela Aparecida Vieira ◽  
Anderson Carlos Faria ◽  
Lucia Vieira

Bacterial cellulose membrane (BCM) is a biomaterial synthesized by bacteria of the genus Gluconocetobacter hansenii with a higher degree of purity than plant cellulose. The commonly used raw material for manipulating bacterial cellulose is kombucha, a beverage consumed by a vast population around the world that promises health benefits. The beverage is composed of tea species Camellia sinenses and a carbon source, refined sucrose, and a starter culture of bacteria and yeast with 10% fermented tea (starter tea) to activate the fermentative process. The Kombucha’s bacterial cellulose membranes (KBCM) are formed over 7 to 10 days on the surface of the fermented product and have the appearance of a gelatinous membrane, this being the by-product of interest. In this work, the objective was to obtain the membrane composed of cellulose via Kombucha and purify it to obtain crystalline cellulose. The purification was performed with distilled water and 0.5M NaOH sodium hydroxide solution to remove residues from the fermentation, successfully removing sugars and bacteria. At the end of the experiments, a lighter film was obtained with coloration close to white, and comparative analyses were performed to verify the structural chemical composition, crystallinity, and morphology of the samples by techniques FTIR, DRX, and SEM, respectively. Then, once the biomaterial was purified, the range of applications expanded to several products to meet the biomedical area, sustainable packaging, and even the fashion industry.


Pharmaceutics ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2063
Author(s):  
Indu Muthancheri ◽  
Rohit Ramachandran

In this study, a hybrid modeling framework was developed for predicting size distribution and content uniformity of granules in a bi-component wet granulation system with components of differing hydrophobicities. Two bi-component formulations, (1) ibuprofen-USP and micro-crystalline cellulose and (2) micronized acetaminophen and micro-crystalline cellulose, were used in this study. First, a random forest method was used for predicting the probability of nucleation mechanism (immersion and solid spread), depending upon the formulation hydrophobicity. The predicted nucleation mechanism probability is used to determine the aggregation rate as well as the initial particle distribution in the population balance model. The aggregation process was modeled as Type-I: Sticking aggregation and Type-II: Deformation driven aggregation. In Type-I, the capillary force dominant aggregation mechanism is represented by the particles sticking together without deformation. In the case of Type-II, the particle deformation causes an increase in the contact area, representing a viscous force dominant aggregation mechanism. The choice between Type-I and II aggregation is determined based on the difference in nucleation mechanism that is predicted using the random forest method. The model was optimized and validated using the granule content uniformity data and size distribution data obtained from the experimental studies. The proposed framework predicted content non-uniform behavior for formulations that favored immersion nucleation and uniform behavior for formulations that favored solid-spreading nucleation.


Author(s):  
Lijuan Gao ◽  
Yaru Su ◽  
Wenxia Song ◽  
Weican Zhang ◽  
Qingsheng Qi ◽  
...  

Cytophaga hutchinsonii is an abundant soil cellulolytic bacterium that uses a unique cellulose degradation mechanism different from those that involve free cellulases or cellulosomes. Though several proteins were identified to be important for cellulose degradation, the mechanism used by C. hutchinsonii to digest crystalline cellulose remains a mystery. In this study, chu_0922 was identified by insertional mutation and gene deletion as an important gene locus indispensable for crystalline cellulose utilization. Deletion of chu_0922 resulted in defect in crystalline cellulose utilization. The Δ 0922 mutant completely lost the ability to grow on crystalline cellulose even with extended incubation, and selectively utilized the amorphous region of cellulose leading to the increased crystallinity. As a protein secreted by the type Ⅸ secretion system (T9SS), CHU_0922 was found to be located on the outer membrane, and the outer membrane localization of CHU_0922 relied on the T9SS. Comparative analysis of the outer membrane proteins revealed that the abundance of several cellulose binding proteins, including CHU_1276, CHU_1277, and CHU_1279, was reduced in the Δ 0922 mutant. Further study showed that CHU_0922 is crucial for the full expression of the gene cluster containing chu_1276 , chu_1277 , chu_1278 , chu_1279 , and chu_1280 ( cel9C ), which is essential for cellulose utilization. Moreover, CHU_0922 is required for the cell surface localization of CHU_3220, a cellulose binding protein that is essential for crystalline cellulose utilization. Our study provides insights into the complex system that C. hutchinsonii uses to degrade crystalline cellulose. IMPORTANCE The widespread aerobic cellulolytic bacterium Cytophaga hutchinsonii , belonging to the phylum Bacteroidetes , utilizes a novel mechanism to degrade crystalline cellulose. No genes encoding proteins specialized in loosening or disruption the crystalline structure of cellulose were identified in the genome of C. hutchinsonii , except for chu_3220 and chu_1557 . The crystalline cellulose degradation mechanism remains enigmatic. This study identified a new gene locus, chu_0922 , encoding a typical T9SS substrate that is essential for crystalline cellulose degradation. Notably, CHU_0922 is crucial for the normal transcription of chu_1276 , chu_1277 , chu_1278 , chu_1279 , and chu_1280 ( cel9C ), which play important roles in the degradation of cellulose. Moreover, CHU_0922 participates in the cell surface localization of CHU_3220. These results demonstrated that CHU_0922 plays a key role in the crystalline cellulose degradation network. Our study will promote the uncovering of the novel cellulose utilization mechanism of C. hutchinsonii.


2021 ◽  
Author(s):  
Dennison Bindhulekha Arya ◽  
Salom Gnana Thanga Vincent ◽  
J.K Reshma ◽  
Junaid Hassan Salahudeen

Abstract Estuarine sediments are best suited for bioprospecting cellulose degrading microorganisms because of continuous input of cellulosic carbon from rivers and terrestrial runoff, and such sediments act as a substrate for decomposition by microbes. Sediment samples were collected from thirteen stations of Ashtamudi estuary, a tropical Ramsar site during April 2016 and January 2017 and analysed for environmental variables such as temperature, pH, electrical conductivity, oxidation- reduction potential, sulphate, total organic carbon (Corg), carbohydrate, protein, lipid and labile organic matter. Microcosm experiments were conducted in the sediment samples to compare native and substrate-induced cellulase enzyme activities in mesophilic and thermophilic conditions added with crystalline cellulose and cellobiose as substrates. Abundance of cellulolytic anaerobes in the roll tubes was higher with cellobiose than crystalline cellulose. Substrate induced enzyme activity was more than native enzyme activity [0.0012±0.0001- 0.004±0.002 (April 2016) and 0.004±0.001- 0.161±0.002 mg glucose h-1 (January 2017)] in the sediment samples and cellulolytic activity was more pronounced in thermophilic conditions during April 2016. Redundancy analysis indicated that salinity was the highest determining factor for explaining variations among bacterial abundance and activity during April 2016 and sediment lipid content during January 2017. The study reveals that estuarine sediments can act as a potential source of thermophilic cellulase enzyme producing bacteria, which needs to be further explored owing to their vast industrial applications.


2021 ◽  
Vol 2069 (1) ◽  
pp. 012012
Author(s):  
Chi Zhang ◽  
Mingyang Chen ◽  
Dominique Derome ◽  
Jan Carmeliet

Abstract Wood is known to swell substantially during moisture adsorption and shrink during desorption. These deformations may lead to wood damage in the form of cracking and disjoining of wooden components in e.g. floor or windows. Two swelling mechanisms may be distinguished: reversible swelling/shrinkage and moisture-induced shape memory effect. In the latter, wood is deformed in the wet state and afterward dried under maintained deformation, in order that wood retains its deformed shape even after the removal of the mechanical loading, called fixation. When wood is wetted again, it loses its fixation, partially regains its original shape, called recovery. These two mechanisms have their origin at the nanoscale and are modelled here using atomistic simulation and after upscaled to continuum level allowing finite element modelling. Hysteretic sorption and swelling are explained at nanoscale by the opening and closing of sorption sites in ad-and desorption, where in desorption water molecules preferentially remained bonded at sorption sites. The moisture-induced shape memory is explained by the moisture-induced activation of the interfaces between the reinforcing crystalline cellulose fibres and its matrix at nanoscale, referred to as a molecular switch. Our work aims to highlight that the understanding of sorption-induced reversible deformation and moisture-induced shape memory may play an important role in wood engineering and in building physics applications.


Author(s):  
Shuaishuai Xie ◽  
Yahong Tan ◽  
Wenxia Song ◽  
Weican Zhang ◽  
Qingsheng Qi ◽  
...  

Cytophaga hutchinsonii is a Gram-negative bacterium belonging to the phylum Bacteroidetes . It digests crystalline cellulose with an unknown mechanism, and possesses a type IX secretion system (T9SS) that can recognize the C-terminal domain (CTD) of the cargo protein as a signal. In this study, the functions of CTD in the secretion and localization of T9SS substrates in C. hutchinsonii were studied by fusing the green fluorescent protein (GFP) with CTD from CHU_2708. CTD is necessary for the secretion of GFP by C. hutchinsonii T9SS. The GFP-CTD CHU_2708 fusion protein was found to be glycosylated in the periplasm with a molecular mass about 5 kDa higher than that predicted from its sequence. The glycosylated protein was sensitive to peptide- N -glycosidase F which can hydrolyze N -linked oligosaccharides. Analyses of mutants obtained by site-directed mutagenesis of asparagine residues in the N-X-S/T motif of CTD CHU_2708 suggest that N -glycosylation occurred on the CTD. CTD N- glycosylation is important for the secretion and localization of GFP-CTD recombinant proteins in C. hutchinsonii . Glycosyltransferase encoding gene chu_3842 , a homologous gene of Campylobacter jejuni pglA , was found to participate in the N -glycosylation of C. hutchinsonii . Deletion of chu_3842 affected cell motility, cellulose degradation, and cell resistance to some chemicals. Our study provided the evidence that CTD as the signal of T9SS was N -glycosylated in the periplasm of C. hutchinsonii . IMPORTANCE The bacterial N -glycosylation system has previously only been found in several species of Proteobacteria and Campylobacterota , and the role of N -linked glycans in bacteria is still not fully understood. C. hutchinsonii has a unique cell-contact cellulose degradation mechanism, and many cell surface proteins including cellulases are secreted by the T9SS. Here, we found that C. hutchinsonii , a member of the phylum Bacteroidetes , has an N -glycosylation system. Glycosyltransferase CHU_3842 was found to participate in the N -glycosylation of C. hutchinsonii proteins, and had effects on cell resistance to some chemicals, cell motility, and cellulose degradation. Moreover, N -glycosylation occurs on the CTD translocation signal of T9SS. The glycosylation of CTD apears to play an important role in affecting T9SS substrates transportation and localization. This study enriched our understanding of the widespread existence and multiple biological roles of N -glycosylation in bacteria.


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