由废弃淡水龙虾壳制备的羧甲基壳聚糖在草莓保鲜过程中的生理作用

利用废弃淡水龙虾壳制备得到的羧甲基壳聚糖应用到草莓的保鲜,为如何有效地解决废弃淡水龙虾壳提供了解决途径.实验结果显示不同浓度的羧甲基壳聚糖处理的草莓腐烂率下降,同时延缓了维生素C水平的下降趋势,其中0.2%浓度的羧甲基壳聚糖的作用效果最为明显.在保鲜过程中,测定草莓果肉中超氧化物歧化酶 (SOD)、过氧化物酶 (POD)、过氧化氢酶(CAT)的活性和脂质过氧化物丙二醛(MDA)的含量的变化,发现经过处理的草莓在保鲜过程中,其SOD,POD,CAT酶活性明显提高,MDA的水平同时显著下降,结果说明羧甲基壳聚糖的保鲜作用可能通过提高抗氧化水平来实现.     

A simple preparation of half N-acetylated chitosan highly soluble in water and aqueous organic solvents

A simple and improved method of preparing highly soluble chitosan (half N-acetylated chitosan) was developed using a series of chitosan samples of low molecular weights, and the solubility of the half N-acetylated chitosan in water and organic solvents was investigated in detail. To reduce the molecular weight, chitosan was treated with NaBO3 under the condition that chitosan was homogeneously dissolved in aqueous acetic acid. Weight-average molecular weights of the obtained chitosan samples were determined using a size-exclusion chromatography system equipped with a low-angle laser light-scattering photometer. Each chitosan sample was then N-acetylated with acetic anhydride under the condition that chitosan was homogeneously dissolved in aqueous acetic acid again. The water solubility of the half N-acetylated chitosan thus prepared increased with decreasing molecular weight. From 1H NMR spectroscopy, it was suggested that the sequence of N-acetylglucosamine and glucosamine residues was random. The solubility of the half N-acetylated chitosan of low molecular weight was rather high even in aqueous dimethylacetamide and dimethylsulfoxide.     

壳聚糖絮凝沉淀纯化翅果油树叶片多糖的方法探究

采用壳聚糖絮凝沉淀法对翅果油树(Elaeagnus mollis Diels)叶片多糖进行纯化研究,分别考察了壳聚糖用量、絮凝时间、絮凝温度及溶液酸碱度(pH)对吸附色素、沉降蛋白质和多糖损失率的影响,并采用正交试验优化出壳聚糖絮凝法纯化翅果油树叶片多糖的最佳条件为:壳聚糖用量1 mg/mL、溶液pH值=5、絮凝时间50~70 min、絮凝温度20~40℃。实验结果表明,在最佳条件下,色素含量和蛋白质含量分别降低了69%左右和35%左右,多糖的纯度也得到了一定的提高。     

Improvement of the yield of physiologically active oligosaccharides in continuous hydrolysis of chitosan using immobilized chitosanases

The continuous production of chitosan oligosaccharides using a packed-bed enzyme reactor was investigated as to the effects of the operation conditions on the yield of pentamers and hexamers of chitosan oligosaccharides. A column reactor packed with immobilized chitosanases prepared by the multipoint attachment method was used for continuous hydrolysis of chitosan. In this reactor, the decrease of the yield of the target intermediate oligosaccharides due to axial mixing was negligible. The surface enzyme density of the support and flow rate of the substrate solution significantly affected the maximum yield of pentamers and hexamers. These effects were summarized as a correlation with the Damk?hler number (Da), defined as the ratio of the maximum reaction rate to the maximum mass transfer rate. The optimum condition was determined based on Da. Under the optimized condition (Da = 0.12), pentamers and hexamers could be produced continuously for a month with a yield of over 35% (7 kg/m(3) in concentration).     

Decoloration of chitosan by UV irradiation

Decoloration of chitosan by UV irradiation, which was used to replace a bleaching step during chitosan preparation, was evaluated under four separate treatments (effect of irradiation time, chitosan/water ratio, stirring speed, and UV light source). The optimal decoloration condition was defined as that producing white chitosan with higher viscosity. Decoloration of chitosan could be achieved effectively using a UV-C light by stirring unbleached chitosan in water (1:8, w/v) for 5 min at 120 rpm. UV irradiation applied under the optimal conditions could be used to produce chitosan with desirable white color (L^* = 76.95, a^* = −0.37, and b^* = 14.04) and high viscosity (1301.7 mPa s at 0.5% w/v in 1.0% v/v acetic acid).     

Practical synthesis and characterization of mannose-modified chitosan

A new and practical laboratory approach to synthesize mannose modified chitosan (Man-chitosan) was developed via reductive amination reaction. Chitosan and mannose were used as raw materials. The reaction condition was mild and controllable. The overall yield was 47-52%. Each reaction products and Man-chitosan were characterized by (1)H NMR, ESI-MS, FT-IR and TGA spectrum. FT-IR and (1)H NMR results showed that mannose conjugated to chitosan via an alkane chain bridge (CH(2)CH(2)). The degree of substitution was calculated by element analysis. TGA results indicated that mannose grafted to chitosan slightly decreased the thermal stability of chitosan in some extent. MTT assay indicated that Man-chitosan was low cytotoxicity against HepG-2 and SMMC-7721 cells.     

Optimal gamma-ray dose and irradiation conditions for producing low-molecular-weight chitosan that retains its chemical structure

This study focuses on the optimal conditions for gamma irradiation to reduce the molecular weight of chitosan but still retain its chemical structure. Chitosan was irradiated under various conditions, i.e. flake solid state (condition 1), flake dispersed in water (condition 2), flake dispersed in 0.05, 0.1, 1 and 2% aqueous K(2)S(2)O(8) solution (conditions 3a, 3b, 3c and 3d, respectively), flake dispersed in 0.5, 1 and 2% aqueous H(2)O(2) solution (conditions 4a, 4b and 4c, respectively), and chitosan acetic acid solution (condition 5). Comparative studies were done using three types of chitosans with molecular weights of the order of 10(5) Da with degrees of deacetylation of 0.80, 0.85 and 0.90%. For all conditions, after irradiation, there were two regions of molecular weight reduction. A severe degradation occurred in the first region with decreases in the molecular weight of 80% for radiation doses up to 50 kGy for conditions 1, 2 and 3 (3a-3c) and 20 kGy for condition 4. In the second region, a slow degradation occurred, which resembled a plateau stage. The results for conditions 3d and 5 were the most dramatic, since the primary structure of chitosan was changed after the irradiation. The degradation of chitosan by gamma rays was found to be most effective for the amorphous structure. The retention of the structure of chitosan after gamma irradiation makes it possible to produce a low-molecular-weight chitosan that retains its functionality, as demonstrated by its activity in the coupling reaction with N,N'-carbonyldiimidazole.     

Preparation and characterization of novel chitosan/gelatin membranes using chitosan hydrogel

Chitin and chitosan are novel biomaterials. The novel chitosan/gelatin membranes were prepared using the suspension of chitosan hydrogel mixed with gelatin. The prepared chitosan/gelatin membranes were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), mechanical, swelling, and thermal studies. The morphology of these chitosan/gelatin membranes was found to be very smooth and homogeneous. The XRD studies showed that the chitosan/gelatin membranes have good compatibility and interaction between the chitosan and gelatin. The stress and elongation of chitosan/gelatin membranes on wet condition showed excellent when the mixture ratio of gelatin was 0.50. The prepared chitosan/gelatin membranes showed good swelling, mechanical and thermal properties. Cell adhesion studies were also carried out using human MG-63 osteoblast-like cells. The cells incubated with chitosan/gelatin membranes for 24 h were capable of forming cell adhesion. Thus the prepared chitosan/gelatin membranes are bioactive and are suitable for cell adhesion suggesting that these membranes can be used for tissue-engineering applications. Therefore, these novel chitosan/gelatin membranes are useful for biomedical applications.     

来源于家蝇、地鳖虫、黄粉虫和河虾壳几丁糖的基本特性

为了了解昆虫来源几丁糖的基本特性及其与河虾来源几丁糖的差别,用同一方法在相同条件下分别以河虾壳(shell of Procambarus clarkii)、家蝇蛹壳(pupa shell ofMusca domestica vicinaMacquart)、地鳖虫壳(shell of Euplyphaga Walker)和黄粉虫蜕(exuviate of Tnebrio molitorL.)为来源制备的几丁糖在灰份、脱乙酰度、分子质量等及红外图谱上进行比较研究。昆虫来源几丁糖,其灰份低于河虾壳来源几丁糖;家蝇蛹壳几丁糖分子质量明显低于其他3种来源的几丁糖;从几丁糖质量角度看,采用家蝇蛹壳和地鳖虫壳可制备脱乙酰度较高的几丁糖;4种来源的几丁糖红外光谱图谱基本一致,具有几丁糖的特征吸收峰。     

Microwave-assisted degradation of chitosan for a possible use in inhibiting crop pathogenic fungi

Degradation of chitosan by H(2)O(2) under microwave irradiation was investigated. The oxidative degradation of chitosan was highly accelerated by microwave irradiation under the condition of low temperature and low concentration of H(2)O(2). The degraded chitosans with low molecular weight (M(w)) were characterized by gel permeation chromatography, Fourier-transform infrared spectroscopy, ultraviolet-visible spectroscopy, X-ray diffraction and elemental analysis. The decrease of M(w) led to transformation of crystal structure and increase of water solubility, whereas no significant chemical structure change in the backbone of chitosan was observed. Antifungal activities of chitosans with different M(w) against crop pathogenic fungi Phomopsis asparagi, Fusarium oxysoporum f. sp. Vasinfectum and Stemphylium solani were investigated at the concentrations of 100, 200 and 400mg/L. All degraded chitosans with low M(w) exhibited enhanced antifungal activity compared with original chitosan and the chitosan of 41.2kDa showed the highest activity. At 400mg/L, the chitosan of 41.2kDa inhibited growth of P. asparagi at 89.3%, stronger than polyoxin and triadimefon, the inhibitory effects of which were found to be 55.5% and 68.5%. All the results indicated that oxidative degradation under microwave irradiation was a promising technique for large-scale production of low M(w) chitosan for use in crop protection.     

壳聚糖作为基因药物载体的研究进展

以壳聚糖及其衍生物作为基因的载体的转染效率受到许多因素的影响, 如复合物粒子大小、壳聚糖/DNA的比值、壳聚糖的分子量、脱乙酰度、转染过程中血清的浓度、介质的pH值等。对壳聚糖进行一定程度的修饰, 可以改变壳聚糖的转染效率。介绍了壳聚糖作为基因转移载体的转染条件, 转染效率和转染机制的研究情况及研究进展。     

Effect of chitosan elicitation and media components on the production of anthraquinone colorants in madder (Rubia akane Nakai) cell culture

To increase the production of anthraquinone colorants in madder (Rubia akane Nakai) cell culture, the effects of elicitation on the colorant production were investigated. Chitosan was the best biotic elicitor among nine plant derived and microbial derived polysaccharides. When elicited with 25 mg/L chitosan, the total production was increased approximately two times in a seven-day culture as compared to that in the unelicited cells. Anthraquinone production was increased in proportion to the contact period up to day 3. Maximum anthraquinone colorants were obtained with 3-day treatment of chitosan. During chitosan elicitation, the total production was increased 1.3 times in MS medium containing galactose as compared to that containing sucrose. The degree of deacetylation in chitosan and the use of growth regulator or addition of precursor did not affect the production of anthraquinone colorants. When madder cells were elicited at optimum condition, anthraquinone concentration and specific anthraquinone content increased 1.3 times (0.69 g/L) and 2.2 times (0.32 g/g DCW), respectively.     

Formation of biocompatible nanoparticles by self-assembly of enzymatic hydrolysates of chitosan and carboxymethyl cellulose

A simple preparation method for biocompatible nanoparticles in high concentration (0.5 wt %) by self-assembly of chitosan and carboxymethyl cellulose hydrolysates was developed. Chitosan and carboxymethyl cellulose were hydrolyzed beforehand with chitosanase and cellulase respectively to make fragments having lower molecular weights. Nanoparticles were spontaneously formed only by mixing the two hydrolysate solutions. The particle size distribution was relatively narrow, about 200 nm in mean size. The mean particle size decreased from 226 nm to 165 nm with decreasing molecular weight of chitosan hydrolysate from 9.5 to 6.8 kDa. The mixing ratio of chitosan and carboxymethyl cellulose hydrolysates also affected particle size. Changes in particle size are discussed in relation to a possible mechanism of polyionic complexation. The chitosan-carboxymethyl cellulose nanoparticles were stably suspended over 1 week even under low pH (pH 3.0), high ionic strength (NaCl 1 M), or low temperature (4 degrees C) conditions.     

Synthesis of methylated chitosan containing aromatic moieties: Chemoselectivity and effect on molecular weight

N-Arylated chitosans were synthesized via Schiff bases formed by the reaction between the primary amino group of chitosan with aromatic aldehydes followed by reduction of the Schiff base intermediates with sodium cyanoborohydride. Treatment of chitosan containing N,N-dimethylaminobenzyl and N-pyridylmethyl substituents with iodomethane under basic conditions led to quaternized N-(4-N,N-dimethylaminobenzyl) chitosan and quaternized N-(4-pyridylmethyl) chitosan. Methylation occurred at either N,N-dimethylaminobenzyl and N-pyridylmethyl groups before the residual primary amino groups of chitosan GlcN units were substituted. The total degree of quaternization of each chitosan varied depending on the extent of N-substitution (ES) and the sodium hydroxide concentration used in methylation. Increasing ES increased the total degree of quaternization but reduced attack at the GlcN units. N,N-dimethylation and N-methylation at the primary amino group of chitosan decreased at higher ES’s. Higher total degrees of quaternization and degrees of O-methylation resulted when higher concentrations of sodium hydroxide were used. The molecular weight of chitosan before and after methylation was determined by gel permeation chromatography under mild acidic condition. The methylation of the N,N-dimethylaminobenzyl derivative with iodomethane was accompanied by numerous backbone cleavages and a concomitant reduction in the molecular weight of the methylated product was observed. The antibacterial activity of water-soluble methylated chitosan derivatives was determined using Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacteria; minimum inhibitory concentrations (MIC) of these derivatives ranged from 32 to 128 μg/mL. The presence of the N,N-dimethylaminobenzyl and N-pyridylmethyl substituents on chitosan backbone after methylation did not enhance the antibacterial activity against S. aureus. However, N-(4-N,N-dimethylaminobenzyl) chitosan with degree of quaternization at the aromatic substituent and the primary amino group of chitosan of 17% and 16–30%, respectively, exhibited a slightly increased antibacterial activity against E. coli.     

Dependence on the Preparation Procedure of the Polymorphism and Crystallinity of Chitosan Membranes

Differences in the polymorphism and crystallinity of chitosan were found in membranes prepared by different procedures when examined by X-ray diffraction measurements for four samples of chitosan differing in the degree of polymerization. When an acetic acid solution of chitosan was dried in air and then soaked in an alkaline solution (method A), both hydrated and anhydrous polymorphs of chitosan were present in the resulting membranes; the latter polymorph made chitosan insoluble in common solvents of chitosan, and its crystallinity increased with decreasing chitosan molecular weight. When a highly concentrated chitosan solution in aqueous acetic acid was neutralized with an alkaline solution (method B), no anhydrous polymorphs were detected in the membrane because of incomplete drying. When aqueous formic acid was used as the solvent, behavior basically similar to that in aqueous acetic acid was observed. In contrast, even with method A, aqueous hydrochloric acid gave a chitosan membrane having very little anhydrous crystallinity. The crystalline polymorph called “1–2”, which has been proposed to be one of four chitosan polymorphs, is considered to be a mixture of hydrated and anhydrous crystals.     

Synthesis and characterization of N-aryl chitosan derivatives

Selective N-arylation of chitosan was performed via a Schiff bases formed by the reaction between the 2-amino group of glucosamine residue of chitosan with an aromatic aldehyde under acidic condition followed by reduction of the Schiff base intermediate with sodium cyanoborohydride (Borch reduction). Aromatic aldehydes bearing either an electron donating or electron withdrawing substituent were used. The chemical structures and thermal properties of the N-aryl chitosans were characterized by FT-IR, (1)H NMR, (13)C NMR, TGA, and DSC. The extent of N-substitution (ES) was influenced by the molar ratio of the aldehyde to the glucosamine residue of chitosan, the reaction time and the substituent on the aromatic ring. Lower ESs resulted from N-arylation using an aldehyde with an electron donating substituent. A linear relationship between the targeted ES and the ES obtained was observed when aldehydes bearing electron withdrawing substituents were employed.     

壳聚糖反渗透膜的制备初探

本文介绍了从虾、蟹甲壳制取壳聚糖用于制备反渗透膜的方法。运用正交试验设计分析制膜条件对膜性能的影响。     

Adsorption properties of gold onto a chitosan derivative

In order to find a material which can be used for the recovery of Au(III), a chitosan derivative was synthesized by carboxymethylation and grafting sulfur groups onto cross-linked chitosan backbone. Adsorption studies were carried out at different pH values to optimize the pH condition. Batch method was conducted to study the effects of parameters such as reaction time, initial metal concentration and temperature on Au(III) sorption. The maximum adsorption affinity for Au(III) was found to be 8.32mmol/g at pH 4.0, 25°C. The results of kinetic study showed that the adsorption reaction followed the pseudo second order model. The derivative showed high adsorption ability and reusability toward Au(III). All results suggested that the chitosan derivative had potential to be utilized in the recovery of Au(III) from aqueous medium.     

Production and characterization of chitosan obtained from <Emphasis Type="Italic">Rhizopus oryzae</Emphasis> grown on potato chip processing waste

Potato chip processing waste of trimmed potato, potato peel and substandard (low-quality) potato chips, obtained from a potato chip processing plant, were used as substrates for chitosan production from Rhizopus oryzae. It was cultured on each waste product at 30 ± 2°C and 70% moisture content for 21 days. Fermented potato peel had the highest yield after 5 days of fermentation. The cultivation condition of chitosan obtained from R. oryzae was optimum for a peel size of less than 6 mesh, 70% moisture content and a pH of 5. Furthermore, the best extraction condition was using 46% sodium hydroxide at 46°C for 13 h followed by 2% acetic acid at 95°C for 8 h. The maximum chitosan yield obtained by these conditions was 10.8 g/kg substrate. Fungal chitosan properties were found to be 86–90% degree of deacetylation, molecular weight of 80–128 kDa and viscosity of 3.1–6.1 mPa s. Therefore, potato peel could be applied as a low cost substrate for chitosan production from R. oryzae.     

Effect of abiotic factors on the antibacterial activity of chitosan against waterborne pathogens

To assess the adaptability of chitosan (from agricultural waste) as a natural disinfectant, its antibacterial activity against bacteria associated with waterborne diseases was investigated by varying such abiotic conditions, as pH and ionic strength and by adding different amounts of acid solvent, metal ions, and EDTA. Two major waterborne pathogens, Escherichia coli and Staphylococcus aureus, were examined. Results showed that organic acids with low carbon number were better solvents for chitosan than were inorganic acids. The effect of pH below 6 on the antibacterial activity of chitosan was significant. The antibacterial activity of chitosan increased with ionic strength but decreased with the addition of metal ions. The addition of Zn(2+) ions inhibited the antibacterial activity of chitosan the most, while the addition of Mg(2+) ions inhibited the antibacterial activity of chitosan the least. This was due to the chelating capacity of chitosan toward metal ions. The antibacterial activity of chitosan against E. coli was enhanced by EDTA. However, the antibacterial activity of chitosan against S. aureus was partially suppressed by EDTA. The antibacterial activity of chitosan was also dependent on its charges and solubility. The antibacterial mechanism of chitosan has currently been hypothesized as being related to surface interference. The results show that the chitosan is a potential bactericide under various environmental conditions.