壳聚糖的抑菌效果研究及其与苯甲酸钠的比较

考察了脱乙酰度和分子量对壳聚糖抑菌效果的影响,结果表明:随着脱乙酰度的增大,其对枯草芽孢杆菌、金黄色葡萄球菌、链球菌和大肠杆菌的抑菌效果增强;而分子量对其抑菌效果的影响则针对供试细菌的不同而有所不同,对枯草芽孢杆菌、金黄色葡萄球菌、链球菌抑制作用最强的分子量范围为50～100kDa,对大肠杆菌的则为10～50kDa.选择最适的抑菌条件,将壳聚糖的抑菌效果与苯甲酸钠进行对比.结果显示:壳聚糖的抑菌效果优于苯甲酸钠,在浓度达到0.25%时即可完全抑制上述四种细菌的生长.     

不同脱乙酰度蚕蛹壳聚糖抑菌性能的比较

研究了3种不同脱乙酰度蚕蛹壳聚糖对9种供试菌的抑菌效果。结果表明:3种壳聚糖对蜡状芽孢杆菌、鼠伤寒杆菌、枯草芽孢杆菌、白色念珠球菌、绿脓假单胞菌、大肠杆菌和金黄色葡萄球菌有明显的抑制作用,对巨大芽孢杆菌和嗜热脂肪芽孢杆菌的抑制作用不明显。随着壳聚糖溶液浓度的增大,其抑菌能力也增强。3种壳聚糖的抑菌活性为:脱乙酰度为95.96%的壳聚糖>脱乙酰度为86.45%的壳聚糖>脱乙酰度为78.12%的壳聚糖。壳聚糖的抑菌活性呈现随pH降低而增加的趋势,当pH值在pH 4.0～5.0,壳聚糖对所有供试菌均能完全抑制。高脱乙酰度蚕蛹壳聚糖作为热加工食品的防腐剂,可稳定保持其抑菌防腐性能。     

小分子羧甲基壳聚糖的抑菌性能研究

采用体外抑菌法分别测定不同小分子量羧甲基壳聚糖对几类菌种的抑制作用。结果表明3 000 u分子量的羧甲基壳聚糖对大肠杆菌、金黄色葡萄球菌、黑曲霉和酵母菌的最低抑菌浓度分别为:0.10%、0.15%、0.15%、0.15%;6 000 u分子量的羧甲基壳聚糖对大肠杆菌、金黄色葡萄球菌、黑曲霉和酵母菌的最低抑菌浓度分别为:0.15%、0.15%、0.20%、0.15%;9 000 u分子量的羧甲基壳聚糖对大肠杆菌、金黄色葡萄球菌、黑曲霉和酵母菌的最低抑菌浓度分别为:0.20%、0.10%、0.20%、0.20%。较高分子量的羧甲基壳聚糖对金黄色葡萄球菌的抑菌效果较好,低分子量的羧甲基壳聚糖对大肠杆菌、黑曲霉、酵母菌的抑菌效果较好。     

壳聚糖对食品中常见菌的抑制研究

为探讨壳聚糖的抑菌影响因素，采用间歇法脱乙酰和双氧水氧化降解法制备了系列壳聚糖产品，对食品中常见细菌金黄色葡萄球菌和大肠肝菌进行定量抑菌实验，考察了氨基含量和分子量对CTS抑菌能力的影响程度。结果显示：脱乙酰度（DD）≥85％，粘均分子量（Mη）≤47万的壳聚糖样品，在浓度达1．0g／L时对两种细菌的抑菌率均能达到100％。壳聚糖的氨基含量对抑菌性能的影响较之分子量因素明显，而分子量的变化在抑菌机制中也起着比较重要的作用。     

几种天然防腐剂的抑菌性能研究

本试验以金黄色葡萄球菌、大肠杆菌、假单胞菌、热死环丝菌为供试验菌,通过滤纸片试验法对常见十种天然防腐剂的抑菌性能进行研究,试验结果显示茶多酚对这四类菌都具有很强的抑菌作用;壳聚糖的抑菌谱也比较宽,但对各菌的抑菌力不如茶多酚强;大蒜对大肠杆菌和金黄色葡萄球菌表现出很强的抑菌作用;溶菌酶、蜂胶和Nisin都主要对G+有较强的抑制作用,而对G-作用弱或无作用;丁香提取液对金黄色葡萄球菌和热死环丝菌的抑菌作用较强;桂皮提取液对金黄色葡萄球菌和大肠杆菌的抑菌作用较强,但对假单胞菌的抑菌作用相对较弱;生姜在试验中未表现出有抑菌作用,迷迭香的抑菌作用也很弱.由此可作为进一步优选几种天然防腐剂复配时的优选依据.     

佛手挥发油抑菌活性的研究

以水蒸气蒸馏法对佛手挥发油进行提取,采用滤纸片法比较了不同提取部位挥发油抑菌作用的大小,测定了佛手果挥发油对各供试菌种的抑菌圈直径,同时就不同浓度及pH下佛手果挥发油的抑菌效果进行初步研究.结果表明:佛手果实挥发油的抑菌效果最好,佛手叶中挥发油只对酵母菌有一定的抑制作用,而枝中挥发油没有抑菌作用.用水蒸气蒸馏法提取果实挥发油得率为1.56%;佛手果实挥发油对酵母菌大肠杆菌、枯草杆菌和金黄色葡萄球菌均有较明显的抑制作用,其中对枯草杆菌的抑菌效果最强,对金黄色葡萄球菌的抑制作用相对较弱;并且挥发油具有很好的热稳定性及抑菌持久性,佛手果挥发油在100%、50%、25%三个浓度上的变化对其抑菌效果的影响明显,且在中性和偏酸性范围内抑菌效果最好.四种供试菌的最低抑菌浓度(MIC)分别为:枯草杆菌1.25%、大肠杆菌1.25%、金黄色葡萄球菌2.5%、酿酒酵母2.5%.     

机械法降解壳聚糖及其体外抗菌性能研究

用搅拌球磨机降解壳聚糖,分别考察了机械研磨时间、温度及搅拌速度对壳聚糖分子量的影响,并用XRD、IR表征机械降解壳聚糖的结构和比较其对金黄色葡萄球菌、大肠杆菌、枯草杆菌的抑菌作用以及对小鼠腹腔巨噬细胞吞噬鸡红细胞能力的影响。结果表明：机械研磨作用使壳聚糖发生了明显的降解,结晶度明显降低,但不对壳聚糖的脱乙酰度产生影响;机械降解壳聚糖对3种菌均有抑菌作用,尤其对金黄色葡萄球菌效果最好,并能提高巨噬细胞吞噬鸡红细胞的能力,增强小鼠细胞的免疫功能。     

丹参提取液体外抑菌活性研究

目的 研究丹参提取液的体外抑菌作用.方法 采用常量肉汤稀释法测定最低抑菌浓度(MIC),探讨其体外抑菌作用.采用杯碟法检测丹参不同提取方式、不同灭菌方式以及改变提取液pH值对金黄色葡萄球菌抑菌活性的影响.结果 丹参醇提液对大肠埃希菌、金黄色葡萄球菌、枯草芽胞杆菌、铜绿假单胞杆菌、短小芽胞杆菌均有抑菌作用,其95%醇提液对上述菌的MIC分别为20,20,40,80,80 mg/mL,对白色念珠菌无抑菌作用.丹参醇提液对大肠埃希菌、金黄色葡萄球菌抑菌活性较强;不同灭菌方式及改变提取液pH值对金黄色葡萄球菌抑菌活性无差异.结论 丹参提取液具有一定抑菌作用.     

无花果叶提取物的抑菌作用研究

为探讨无花果叶逐级提取物的抑菌作用,实验测定了无花果叶逐级提取物对各实验菌种的抑菌圈及最低抑菌浓度,结果表明各级提取物对实验菌分别有不同程度的抑菌作用,其中对金黄色葡萄球菌和枯草杆菌的抑菌作用最为明显,各级提取物的抑菌强度依次为:乙酸乙酯萃取物(金黄色葡萄球菌:MIC≤6.25mg/mL,枯草杆菌MIC:≤12.5mg/mL)、正丁醇萃取液(两种菌的抑菌作用:MIC≤25mg/mL)、石油醚萃取液(两种菌的抑菌作用:MIC≤50mg/mL),其中乙酸乙酯萃取物的抑菌活性最强。     

丹参提取液体外抑菌活性研究

目的 研究丹参提取液的体外抑菌作用.方法 采用常量肉汤稀释法测定最低抑菌浓度(MIC),探讨其体外抑菌作用.采用杯碟法检测丹参不同提取方式、不同灭菌方式以及改变提取液pH值对金黄色葡萄球菌抑菌活性的影响.结果 丹参醇提液对大肠埃希菌、金黄色葡萄球菌、枯草芽胞杆菌、铜绿假单胞杆菌、短小芽胞杆菌均有抑菌作用,其95%醇提液对上述菌的MIC分别为20,20,40,80,80 mg/mL,对白色念珠菌无抑菌作用.丹参醇提液对大肠埃希菌、金黄色葡萄球菌抑菌活性较强;不同灭菌方式及改变提取液pH值对金黄色葡萄球菌抑菌活性无差异.结论 丹参提取液具有一定抑菌作用.     

Antibacterial activity of chitosans and chitosan oligomers with different molecular weights

Antibacterial activities of six chitosans and six chitosan oligomers with different molecular weights (Mws) were examined against four gram-negative (Escherichia coli, Pseudomonas fluorescens, Salmonella typhimurium, and Vibrio parahaemolyticus) and seven gram-positive bacteria (Listeria monocytogenes, Bacillus megaterium, B. cereus, Staphylococcus aureus, Lactobacillus plantarum, L. brevis, and L. bulgaricus). Chitosans showed higher antibacterial activities than chitosan oligomers and markedly inhibited growth of most bacteria tested although inhibitory effects differed with Mws of chitosan and the particular bacterium. Chitosan generally showed stronger bactericidal effects with gram-positive bacteria than gram-negative bacteria in the presence of 0.1% chitosan. The minimum inhibitory concentration (MIC) of chitosans ranged from 0.05% to >0.1% depending on the bacteria and Mws of chitosan. As a chitosan solvent, 1% acetic acid was effective in inhibiting the growth of most of the bacteria tested except for lactic acid bacteria that were more effectively suppressed with 1% lactic or formic acids. Antibacterial activity of chitosan was inversely affected by pH (pH 4.5-5.9 range tested), with higher activity at lower pH value.     

Preparation and characterisation of selected physicochemical and functional properties of β‐chitosans from squid pen

Squid pen β‐chitosans prepared under various deacetylation conditions (30%, 35%, 40% and/or 45% NaOH for 15, 30 and/or 60 min) were characterised. β‐Chitosans (deacetylated with 35–45% NaOH for 15–60 min) had 87.1–96.2% degree of deacetylation (DD), 93.5–96.7% solubility and 120.5–654.9 mPa s viscosity. Treatment with 30% NaOH for 15–60 min yielded inadequately deacetylated β‐chitosans (DD = 51.9–80.2%). Two chitosans prepared under 35% NaOH for 15 min and 45% NaOH for 30 min (designated as 35%–15 and 45%–30, respectively) were further compared. Drying (sun‐drying vs. oven‐drying) methods did not affect DD. 35%–15 chitosan exhibited lower nitrogen, DD and bulk density, but higher viscosity compared with 45%–30 chitosan. Higher water‐ and fat‐binding capacity but lower DPPH radical scavenging activity were observed for 35%–15 chitosan compared with 45%–30 chitosan. Compared with 45%–30 chitosan, 35%–15 chitosan exhibited higher antibacterial activity against Salmonella Enteritidis and Listeria monocytogenes, but lower antibacterial activity against Escherichia coli.     

Antimicrobial effects of chitosans and chitooligosaccharides, upon Staphylococcus aureus and Escherichia coli, in food model systems

The objective of this study was to elucidate the controversial relationship between the molecular weight (MW) of chitosans and their antibacterial activity (upon different inoculum levels, at several concentrations). The influence of food components on the activity was also ascertained, as well as acceptance by a sensory panel. All the compounds tested exhibited antibacterial activity against Staphylococcus aureus and Escherichia coli. This activity was shown to be closely dependent on the inoculum level, MW and concentration used. Within 4h at 10(3) cells/mL, all five compounds, at every concentration (0.5%, 0.25% and 0.1%, w/v), proved to be bactericidal; for higher inocula, 0.1% (w/v) was only bacteriostatic; at 10(7) or 10(5) cells/mL, and independently of the inoculum level, 0.25% (w/v) of any chitooligosaccharide (COS) mixture was sufficient to reduce the E. coli initial population by at least 3 log cycles; COS never exhibited bactericidal action over S. aureus, unlike high and medium MW chitosans-which, at 0.5% (w/v), presented a bactericidal effect even against 10(7) cells/mL. When incorporated in liquid food matrices, medium and high MW chitosans maintained their activity, for both matrices and bacteria, although a slower activity was noticeable in milk; however, COS lost their activity upon both bacteria in milk after 4-8h. Furthermore, addition of chitosans to apple juice led to several unpleasant off-flavors, such as astringency and after taste--which increased in magnitude with MW.     

Antibacterial effects of chitosan solution against Legionella pneumophila, Escherichia coli, and Staphylococcus aureus

Chitosan has been shown to have antibacterial activities on the growth of a wide variety of bacteria. Chitosan solution has been sold commercially for use as an antibacterial agent. Chitosan solution contains not only chitosan but also organic acids as solvents and desalted Japan Sea Proper Water (dJSPW). We aimed to clarify whether chitosan solution has antibacterial activity against bacteria invading bath water, and then to explore the causative factor among these ingredients. The antibacterial activity of full-strength chitosan solution and of 10(2)- and 10(4)-fold chitosan solution diluted with purified water was studied against Legionella pneumophila serogroups 1 (L. pneumophila SG1) and 6 (L. pneumophila SG6), Escherichia coli (E. coli), and Staphylococcus aureus (S. aureus) for 7 days at 37 degrees C. To clarify the causative factor in the antibacterial activity against E. coli, the antibacterial activities of the full-strength and diluted chitosan solutions for 24 h were examined. L. pneumophila SG1 and SG6, and E. coli could not survive in the chitosan solution or in the 10(2)-fold dilute solution for over a day at 37 degrees C. The cells of S. aureus were found to have decreased more than 2.46 log cfu/ml after 1 day of incubation, not only in the chitosan solutions, but also in phosphate buffer solution as a control. No inhibitory effect of dJSPW on the growth of the bacteria was observed. The antibacterial activity of the chitosan solution was lower compared with those of the organic acids solutions, and it increased with decreasing pH value. We observed the antibacterial activity of chitosan solution against L. pneumophila SG1 and SG6, and E. coli, suggesting it may be due to the decreased pH value derived from organic acids rather than from chitosan itself or dJSPW.     

壳聚糖抑菌作用位点的研究

采用异硫氰酸荧光素标记3种分子质量(3kD、300～400kD与700kD)壳聚糖,体外检测荧光标记壳聚糖的抑菌活性,并运用激光共聚焦显微镜研究荧光标记壳聚糖对大肠杆菌的抑菌作用位点。结果表明:荧光标记壳聚糖依然具有抑菌活性,但相对未标记壳聚糖而言,3种壳聚糖的抑菌活性分别减小了4.22%、19.56%和14.54%。小分子质量(3kD)壳聚糖能进入细菌内部,其抑菌位点可能为细菌细胞内某物质,大分子质量(300～400kD和700kD)壳聚糖则不能进入细菌内部,其抑菌位点可能为细菌细胞膜。     

壳聚糖的抗菌性研究进展与抗菌纺织品开发

 为深入研究壳聚糖（CS）在抗菌纺织品上的应用,对CS的抗菌机制及其抗菌纺织品开发现状进行综述。介绍了分子量、脱乙酰度、溶液浓度、pH值、溶剂种类、衍生化等因素与抗菌性的关系。结合实际应用认为,CS要发挥优良的抗菌性,应具有高脱乙酰度、中等分子质量,以乙酸作为溶剂,pH值略高于4即可,同时其质量浓度在2.5g/L以上,还可通过衍生化反应进一步提高其抗菌性。分析CS抗菌纺织品开发现状得知,目前主要通过对传统织物固着CS后整理和开发CS纤维制品这2种方式实现纺织品抗菌,前者抗菌性会随产品洗涤次数增加而逐渐降低,而后者将是较好的解决办法,但还有待进一步研究。     

Antibacterial activity of N-alkylated disaccharide chitosan derivatives

Antibacterial activity of the water-soluble N-alkylated disaccharide chitosan derivatives against Escherichia coli and Staphylococcus aureus was investigated. It was found that the antibacterial activity of chitosan derivatives was affected by the degree of substitution (DS) with disaccharide and the kind of disaccharide present in the molecule. Regardless the kind of disaccharide linked to the chitosan molecule, a DS of 30-40%, in general, exhibited the most pronounced antibacterial activity against both test organisms. E. coli and S. aureus were most susceptible to cellobiose chitosan derivative DS 30-40% and maltose chitosan derivative DS 30-40%, respectively, among the various chitosan derivatives examined. Although the disaccharide chitosan derivatives showed less antibacterial activity than the native chitosan at pH 6.0, the derivatives exhibited a higher activity than native chitosan at pH 7.0. Antibacterial activity of the chitosan derivatives (DS 30-40%) against E. coli increased as the pH increased from 5.0 and reached a maximum around the pH of 7.0-7.5. The effect of pH on the antibacterial activity of chitosan derivatives against S. aureus was not as pronounced as that observed with E. coli. Population reduction of E. coli or S. aureus in nutrient broth increased markedly upon increasing the concentration of chitosan derivatives from 0 to 500 ppm. No marked increase in population reduction was noted with further increase in the concentration of chitosan derivatives even up to 2000 ppm.     

漆酶催化香草醛接枝壳聚糖及其在抗菌纸中的应用

以香草醛(VA)为底物,利用漆酶作为催化剂催化香草醛与壳聚糖接枝反应。对VA-壳聚糖接枝产物分别进行线性电位滴定、红外光谱检测,确定产物接枝率和结构的变化,对其抗氧化性和抑菌性能进行分析并采用表面涂布法和浆内添加法制备抗菌纸。结果表明,VA-壳聚糖的接枝率约为10.2%;红外光谱分析表明氨基的吸收峰强度减弱,且在1637 cm-1处出现新的吸收峰,说明在漆酶催化作用下香草醛与壳聚糖上的氨基参加反应形成席夫碱(CN)结构;VA-壳聚糖对ABTS·+自由基半数抑制浓度(IC50)值为1.62 mg/mL,还原能力检测得出浓度为5 mg/mL时在700 nm处的吸光度较原样提高0.321,表明VA-壳聚糖的抗氧化性能得到提高;VA-壳聚糖的抑菌性能较壳聚糖原样有所提高。采用表面涂布法和浆内添加法制备VA-壳聚糖抗菌纸,可以赋予纸张良好的抑菌性能,同时可以改善纸张的物理性能。     

Antibacterial activity of chemically defined chitosans: influence of molecular weight, degree of acetylation and test organism

Chitosans, polysaccharides obtained from the exoskeleton of crustaceans, have been shown to exert antibacterial activity in vitro and their use as a food preservative is of growing interest. However, beyond a consensus that chitosan appears to disrupt the bacterial cell membrane, published data are inconsistent on the chemical characteristics that confer the antibacterial activity of chitosan. While most authors agree that the net charge density of the polymer (reflected in the fraction of positively charged amino groups at the C-2 position of the glucosamine unit) is an important factor in antibacterial activity, conflicting data have been reported on the effect of molecular weight and on the susceptibility among different bacterial species to chitosan. Therefore, we prepared batches of water-soluble hydrochloride salts of chitosans with weight average molecular weights (M_w) of 2-224 kDa and degree of acetylation of 0.16 and 0.48. Their antibacterial activity was evaluated using tube inhibition assays and membrane integrity assays (N-Phenyl-1-naphthylamine fluorescence and potassium release) against Bacillus cereus, Escherichia coli, Salmonella Typhimurium and three lipopolysaccharide mutants of E. coli and S. Typhimurium. Chitosans with lower degree of acetylation (F_A = 0.16) were more active than the more acetylated chitosans (F_A = 0.48). No trends in antibacterial action related to increasing or decreasing M_w were observed although one of the chitosans (M_w 28.4 kDa, F_A = 0.16) was more active than the other chitosans, inhibiting growth and permeabilizing the membrane of all the test strains included. The test strains varied in their susceptibility to the different chitosans with wild type S. Typhimurium more resistant than the wild type E. coli. Salmonellae lipopolysaccharide mutants were more susceptible than the matched wild type strain. Our results show that the chitosan preparation details are critically important in identifying the antibacterial features that target different test organisms.