Preparative methods of phosphorylated chitin and chitosan--an overview

Biomaterials such as chitin, chitosan and their derivatives have a significant and rapid development in recent years. Chitin and chitosan have become cynosure of all party because of an unusual combination of biological activities plus mechanical and physical properties. However, the applications of chitin and chitosan are limited due to its insolubility in most of the solvents. The chemical modification of chitin and chitosan are keen interest because of these modifications would not change the fundamental skeleton of chitin and chitosan but would keep the original physicochemical and biochemical properties. They would also bring new or improved properties. The chemical modification of chitin and chitosan by phosphorylation is expected to be biocompatible and is able to promote tissue regeneration. In view of rapidly growing interest in chitin and chitosan and their chemical modified derivatives, we are here focusing the recent developments on preparation of phosphorylated chitin and chitosan in different methods.     

Chitosan derivatives obtained by chemical modifications for biomedical and environmental applications

Chitosan is a natural based polymer, obtained by alkaline deacetylation of chitin, which presents excellent biological properties such as biodegradability and immunological, antibacterial and wound-healing activity. Recently, there has been a growing interest in the chemical modification of chitosan in order to improve its solubility and widen its applications. The main chemical modifications of chitosan that have been proposed in the literature are reviewed in this paper. Moreover, these chemical modifications lead to a wide range of derivatives with a broad range of applications. Recent and relevant examples of the distinct applications, with particular emphasis on tissue engineering, drug delivery and environmental applications, are presented.     

A new method for the quantification of chitin and chitosan in edible mushrooms

Along with β-glucans, chitin is the dominant component of the fungal cell wall. Chitosan, the deacetylated form of chitin, has found quite a number of biomedical and biotechnological applications recently. Mushroom chitin could be an important source for chitosan production. A direct determination of chitin and chitosan in mushrooms is of expedient interest. In this paper, a new method for the quantification of chitin and chitosan is described. This method is based on the specific reaction between polyiodide anions and chitosan and on measuring the optical density of the insoluble polyiodide–chitosan complex. After deacetylation, chitin can also be quantified. The specificity of the reaction is used to quantify the polymers in the presence of complex matrices. With this new spot assay, the chitin content of mycelia and fruiting bodies from several basidiomycetes and an ascomycete were analysed. The presented method could also be used for the determination in other samples as well. The chitin content of the analysed species varies between 0.4 and 9.8 g chitin per 100 g of dry mass. Chitosan could not be detected in our mushroom samples, indicating that the glucosamine units are mostly acetylated.     

Chitin deacetylase product inhibition

Chitin deacetylase is the only known enzyme catalyzing the hydrolysis of the acetamino linkage in the N-acetylglucosamine units of chitin and chitosan. This reaction can play an important role in enzymatic production of chitosan from chitin, or in enzymatic modification of chitosan, which has applications in medicine, pharmacy or plant protection. It was previously shown that acetic acid, a product of the deacetylation process, may act as an inhibitor of chitin deacetylase. Here we show the mechanism of inhibition of chitin deacetylase isolated from Absidia orchidis vel coerulea by acetic acid released during the deacetylation process. The process follows competitive inhibition with respect to acetic acid with an inhibition constant of K(i) = 0.286 mmol/L. These results will help to find the optimal system to carry out the enzymatic deacetylation process for industrial applications.     

Chitin and chitosan biopolymer production from the Iranian medicinal fungus Ganoderma lucidum: Optimization and characterization

Abstract

Chitin and chitosan with unique properties and numerous applications can be produced from fungus. The production of chitin and chitosan from the mycelia of an Iranian Ganoderma lucidum was studied to improve cell growth and chitin productivity. Inoculum size and initial pH as two effective variables on the growth of G. lucidum and chitin production were optimized using response surface method (RSM) by central composite design (CCD). The results verified the significant effect of these two variables on the cell growth and chitin production. In optimum conditions, including pH?=?5.7 and inoculum size of 7.4%, the cell dry weight was 5.91?g/L and the amount of chitin production was 1.08?g/L with the productivity of 0.083?g/(L day). The produced chitin and chitosan were characterized using XRD and FTIR. Moreover, the antibacterial activity of the produced chitosan was investigated and compared with the commercial chitosan. The results showed that the produced chitin and chitosan had suitable quality and the Iranian G. lucidum would be a great source for safe and high-quality chitin and chitosan production.     

Anti-inflammatory effect of chemically modified chitin

Anti-inflammatory effects of the three types of chitin derivatives namely phosphated chitin (P-chitin), phosphated–sulfated chitin (PS-chitin), and sulfated chitin (S-chitin) were investigated using a canine model of chitosan-induced pneumonia. After simultaneous administration of chitosan with or without each chitin derivative (chitosan alone: n=6, chitosan and P-chitin: n=6, chitosan and PS-chitin: n=1, and chitosan and S-chitin: n=3), hematological examination and X-ray image processing were performed for up to 24 h. Then the lungs were recovered and were evaluated by softex imaging after inflation and fixation. The hematological findings showed that PS-chitin and S-chitin did not prevent the decrease in white blood cell (WBC) count as seen in dogs administered chitosan, while P-chitin prevented such decrease in WBC count. The surface of the inflated and fixed lung specimens was hemorrhagic in the PS- and S-chitin groups as well as in the chitosan group, while the lung looked like normal in the P-chitin group. The pulmonary blood vessels of the chitosan group showed severe change while the P-chitin group showed no changes with softex findings. Furthermore, the pattern of histogram density obtained with image processing of thoracic X-ray in P-chitin group did not change among pre and post administration while chitosan group showed rightward movement and significant changes on parameters. The cause of which is attribured to an attenuation of X-ray permeability by angiectasis of the lung.     

Novel chitin and chitosan nanofibers in biomedical applications

Chitin and its deacetylated derivative, chitosan, are non-toxic, antibacterial, biodegradable and biocompatible biopolymers. Due to these properties, they are widely used for biomedical applications such as tissue engineering scaffolds, drug delivery, wound dressings, separation membranes and antibacterial coatings, stent coatings, and sensors. In the recent years, electrospinning has been found to be a novel technique to produce chitin and chitosan nanofibers. These nanofibers find novel applications in biomedical fields due to their high surface area and porosity. This article reviews the recent reports on the preparation, properties and biomedical applications of chitin and chitosan based nanofibers in detail.     

Biomaterials based on chitin and chitosan in wound dressing applications

Wound dressing is one of the most promising medical applications for chitin and chitosan. The adhesive nature of chitin and chitosan, together with their antifungal and bactericidal character, and their permeability to oxygen, is a very important property associated with the treatment of wounds and burns. Different derivatives of chitin and chitosan have been prepared for this purpose in the form of hydrogels, fibers, membranes, scaffolds and sponges. The purpose of this review is to take a closer look on the wound dressing applications of biomaterials based on chitin, chitosan and their derivatives in various forms in detail.     

A novel soft and cotton-like chitosan-sugar nanoscaffold

A novel type of chitosan nanoscaffold with a soft and cotton-like appearance is proposed. The key to success is based on two points: (i) the change in morphology of chitin whisker to chitosan nanoscaffold and (ii) the surface modification of the nanoscaffold chitosan with a sugar unit. Simple deacetylation of chitin whisker gives a colloidal solution of chitosan, of which the chitosan is in a nanoscaled scaffold. Surface functionalization of the chitosan nanoscaffold with lactose or maltose via a heterogeneous system in water at room temperature results in a soft and cotton-like chitosan containing mesopores. As all steps are organic solvent free, this chitosan-sugar nanoscaffold might be a promising material for biopolymer-supported tissue engineering.     

Chitosan functional properties

Chitosan is a partially deacetylated polymer of N-acetyl glucosamine. It is essentially a natural, water-soluble, derivative of cellulose with unique properties. Chitosan is usually prepared from chitin (2 acetamido-2-deoxy β-1,4-D-glucan) and chitin has been found in a wide range of natural sources (crustaceans, fungi, insects, annelids, molluscs, coelenterata etc.) However chitosan is only manufactured from crustaceans (crab and crayfish) primarily because a large amount of the crustacean exoskeleton is available as a by product of food processing. Squid pens (a waste byproduct of New Zealand squid processing) are a novel, renewable source of chitin and chitosan. Squid pens are currently regarded as waste and so the raw material is relatively cheap. This study was intended to assess the functional properties of squid pen chitosan. Chitosan was extracted from squid pens and assessed for composition, rheology, flocculation, film formation and antimicrobial properties. Crustacean chitosans were also assessed for comparison. Squid chitosan was colourless, had a low ash content and had significantly improved thickening and suspending properties. The flocculation capacity of squid chitosan was low in comparison with the crustacean sourced chitosans. However it should be possible to increase the flocculation capacity of squid pen chitosan by decreasing the degree of acetylation. Films made with squid chitosan were more elastic than crustacean chitosan with improved functional properties. This high quality chitosan could prove particularly suitable for medical/analytical applications. This revised version was published online in November 2006 with corrections to the Cover Date.     

壳聚糖基温敏水凝胶的研究进展

壳聚糖是一种由甲壳素脱乙酰化得到的氨基多糖,具有生物相容性、低细胞毒性和可生物降解性等特点。壳聚糖/β-甘油磷酸钠溶液温敏水凝胶在组织工程、药物缓释等领域多有报道,其成胶性能取决于凝胶的组分和浓度。针对单纯壳聚糖水凝胶强度较低、降解较快、药物突释等缺陷,通常对壳聚糖进行改性或引入新材料共混,获得更符合实际需要的壳聚糖基温敏水凝胶。对近年来壳聚糖基水凝胶的研究进展进行综述,包括改性壳聚糖、共混体系等,概述了其在组织工程(软骨、血管、神经修复)、药物缓释(癌症药物缓释、糖尿病治疗)领域中研究和应用的新进展,以期为后续温敏水凝胶的进一步研究提供参考。     

The co-ordination of chitosan and chitin synthesis in Mucor rouxii

Chitin synthetase preparations from cell walls and chitosomes of the fungus Mucor rouxii were tested for their ability to synthesize chitosan when incubated with uridine diphosphate N-acetyl-D-glucosamine in the presence of chitin deacetylase. The most effective chitin synthetase preparation was one dissociated from cell walls with digitonin. The rate of chitosan synthesis by the wall-dissociated chitin synthetase was about three times that of an equivalent amount of cell walls. The chitosan-synthesizing ability of chitosomes was relatively low, but was more than tripled by treatment with digitonin. Presumably, digitonin improves chitosan yields of dissociating chitin synthetase. The dissociated enzyme would produce dispersed chitin chains that could be attacked by chitin deacetylase before they have time to crystallize into microfibrils. The regulation of chitin and chitosan syntheses in vivo may be determined by the organization of chitin synthetase molecules at the cell surface. Those molecules that remain organized as a complex, similar if not identical to that found in chitosomes, would produce mainly chitin. Chitosan would be preferentially produced by chitin synthetase molecules which are dispersed upon reaching the cell surface.     

Membrane chromatography: preparation and applications to protein separation

As a result of the convective flow of solutes through porous membranes, membrane chromatography has a higher capture efficiency and a higher productivity than column chromatography and shows most promising industrial applications for the recovery, isolation, and purification of proteins and enzymes. This paper presents a comprehensive review of the methods for preparation of adsorptive membranes (such as surface modification, in situ copolymerization, direct formation from hydrophilic materials, and functionalized particulate-entrapped membranes) and deals particularly with novel macroporous chitin and chitosan membranes for protein separations developed by the authors.     

Chitosan but not chitin activates the inflammasome by a mechanism dependent upon phagocytosis

Chitin is an abundant polysaccharide found in fungal cell walls, crustacean shells, and insect exoskeletons. The immunological properties of both chitin and its deacetylated derivative chitosan are of relevance because of frequent natural exposure and their use in medical applications. Depending on the preparation studied and the end point measured, these compounds have been reported to induce allergic responses, inflammatory responses, or no response at all. We prepared highly purified chitosan and chitin and examined the capacity of these glycans to stimulate murine macrophages to release the inflammasome-associated cytokine IL-1β. We found that although chitosan was a potent NLRP3 inflammasome activator, acetylation of the chitosan to chitin resulted in a near total loss of activity. The size of the chitosan particles played an important role, with small particles eliciting the greatest activity. An inverse relationship between size and stimulatory activity was demonstrated using chitosan passed through size exclusion filters as well as with chitosan-coated beads of defined size. Partial digestion of chitosan with pepsin resulted in a larger fraction of small phagocytosable particles and more potent inflammasome activity. Inhibition of phagocytosis with cytochalasin D abolished the IL-1β stimulatory activity of chitosan, offering an explanation for why the largest particles were nearly devoid of activity. Thus, the deacetylated polysaccharide chitosan potently activates the NLRP3 inflammasome in a phagocytosis-dependent manner. In contrast, chitin is relatively inert.     

Chitin whiskers: an overview

Chitin is the second most abundant semicrystalline polysaccharide. Like cellulose, the amorphous domains of chitin can also be removed under certain conditions such as acidolysis to give rise to crystallites in nanoscale, which are the so-called chitin nanocrystals or chitin whiskers (CHWs). CHW together with other organic nanoparticles such as cellulose whisker (CW) and starch nanocrystal show many advantages over traditional inorganic nanoparticles such as easy availability, nontoxicity, biodegradability, low density, and easy modification. They have been widely used as substitutes for inorganic nanoparticles in reinforcing polymer nanocomposites. The research and development of CHW related areas are much slower than those of CW. However, CHWs are still of strategic importance in the resource scarcity periods because of their abundant availability and special properties. During the past decade, increasing studies have been done on preparation of CHWs and their application in reinforcing polymer nanocomposites. Some other applications such as being used as feedstock to prepare chitosan nanoscaffolds have also been investigated. This Article is to review the recent development on CHW related studies.     

Chitin and Chitosan: Functional Biopolymers from Marine Crustaceans

Chitin and chitosan, typical marine polysaccharides as well as abundant biomass resources, are attracting a great deal of attention because of their distinctive biological and physicochemical characteristics. To fully explore the high potential of these specialty biopolymers, basic and application researches are being made extensively. This review deals with the fundamental aspects of chitin and chitosan such as the preparation of chitin and chitosan, crystallography, extent of N-acetylation, and some properties. Recent progress of their chemistry is then discussed, focusing on elemental modification reactions including acylation, alkylation, Schiff base formation and reductive alkylation, carboxyalkylation, phthaloylation, silylation, tosylation, quaternary salt formation, and sulfation and thiolation.     

甲壳素/壳聚糖在酶固定化中的应用

作为功能性材料,甲壳素与壳聚糖分布广泛,且具有一系列独特的性质:无毒性、凝胶性、生物适应性、降解产物的无毒性、显著的蛋白质亲和性等。正是由于这些特性,虽然甲壳素/壳聚糖材料目前尚未得到充分的开发利用,但是与其它一些酶的固定化载体相比,具有广泛的开发前景。该文综述了近年来甲壳素/壳聚糖在酶的固定化方面的一些研究成果。主要包括:甲壳素/壳聚糖的理化性质、载体不同制备方法的特色和差异、在食品工业、非食品工业、环保、酶的分离纯化以及医疗应用方面的研究进展。     

A new route for chitosan immobilization onto polyethylene surface

Low-density polyethylene (LDPE) belongs to commodity polymer materials applied in biomedical applications due to its favorable mechanical and chemical properties. The main disadvantage of LDPE in biomedical applications is low resistance to bacterial infections. An antibacterial modification of LDPE appears to be a solution to this problem. In this paper, the chitosan and chitosan/pectin multilayer was immobilized via polyacrylic acid (PAA) brushes grafted on the LDPE surface. The grafting was initiated by a low-temperature plasma treatment of the LDPE surface. Surface and adhesive properties of the samples prepared were investigated by surface analysis techniques. An antibacterial effect was confirmed by inhibition zone measurements of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The chitosan treatment of LDPE led to the highest and most clear inhibition zones (35mm(2) for E. coli and 275mm(2) for S. aureus).     

serpentine and vermiform encode matrix proteins with chitin binding and deacetylation domains that limit tracheal tube length in Drosophila

Many organs contain epithelial tubes that transport gases or liquids . Proper tube size and shape is crucial for organ function, but the mechanisms controlling tube diameter and length are poorly understood. Recent studies of tracheal (respiratory) tube morphogenesis in Drosophila show that chitin synthesis genes produce an expanding chitin cylinder in the apical (luminal) extracellular matrix (ECM) that coordinates the dilation of the surrounding epithelium . Here, we describe two genes involved in chitin modification, serpentine (serp) and vermiform (verm), mutations in which cause excessively long and tortuous tracheal tubes. The genes encode similar proteins with an LDL-receptor ligand binding motif and chitin binding and deacetylation domains. Both proteins are expressed and secreted during tube expansion and localize throughout the lumen in a chitin-dependent manner. Unlike previously characterized chitin pathway genes, serp and verm are not required for chitin synthesis or secretion but rather for its normal fibrillar structure. The mutations also affect structural properties of another chitinous matrix, epidermal cuticle. Our work demonstrates that chitin and the matrix proteins Serp and Verm limit tube elongation, and it suggests that tube length is controlled independently of diameter by modulating physical properties of the chitin ECM, presumably by N-deacetylation of chitin and conversion to chitosan.     

Chitin deacetylases: new, versatile tools in biotechnology

Chitin deacetylases have been identified in several fungi and insects. They catalyse the hydrolysis of N-acetamido bonds of chitin, converting it to chitosan. Chitosans, which are produced by a harsh thermochemical procedure, have several applications in areas such as biomedicine, food ingredients, cosmetics and pharmaceuticals. The use of chitin deacetylases for the conversion of chitin to chitosan, in contrast to the presently used chemical procedure, offers the possibility of a controlled, non-degradable process, resulting in the production of novel, well-defined chitosan oligomers and polymers.