紫球藻及其胞外多糖对小鼠免疫功能的调节作用

探讨紫球藻及其胞外多糖对免疫低下小鼠免疫功能的调节作用,用环磷酰胺(CY)建立昆明种小鼠免疫功能低下的实验模型。小鼠随机均分6组,其中2组每天分别给予紫球藻藻粉1200、600 mg/kg灌胃,2组给予紫球藻胞外多糖300、150 mg/kg灌胃,正常对照组及CY组用等容积生理盐水灌胃,连续14 d。实验结果表明紫球藻及其胞外多糖均可显著提高免疫抑制小鼠的脾指数、胸腺指数、碳廓清能力、单核细胞吞噬功能,并可对抗环磷酰胺引起的外周血白细胞数目下降,因此紫球藻藻粉及其胞外多糖对小鼠免疫功能具有一定的正向调节作用。毒性实验表明,小鼠腹腔注射300、150 mg/kg紫球藻胞外多糖溶液和分别给予紫球藻藻粉1200、600 mg/kg灌胃,小鼠未出现死亡,也未有明显毒性反应出现的一些指标变化,说明紫球藻藻粉及胞外多糖的安全性较高。     

紫球藻胞外多糖抗呼吸道合胞病毒(RSV)活性研究

采用体外细胞培养的方法,在Hela细胞系上检测了来自紫球藻的胞外多糖及其组分的抗呼吸道病毒(RSV)活性。发现紫球藻胞外多糖对呼吸道合胞病毒具有强烈的抑制活性,同时对宿主细胞的抑制作用很小。分离组分中的强带电性组分ESPSⅥ活性最高,其TI值达3125,为阳性对照药病毒唑的40余倍。     

通入CO2和补加氮源对紫球藻生长和代谢物质积累的影响

论文研究了柱状光生物反应器中CO2浓度和氮源添加模式等培养条件对紫球藻(Porphyridium sp.UTEX 637)生长和主要代谢产物积累的影响。结果表明,通入CO2能显著促进紫球藻的生长及胞外多糖、水溶性蛋白和总脂等生物活性物质的积累,其中1%的CO2浓度对紫球藻的生长及活性物质积累最有利,该条件下紫球藻的最高干重可达到8.14 g/L,是对照组的4.93倍;胞外多糖产量明显高于对照组,最高可达到238.8 mg/L;补加氮源KNO3对紫球藻生长及胞外多糖、水溶性蛋白含量均有明显的促进作用,最高干重、胞外多糖含量可分别达到对照组的1.5倍和1.25倍,但不利于总脂的积累。     

脱落酸对紫球藻（Porphyridiu sp.）生长代谢的影响

为优化培养条件和提高紫球藻(Porphyridiu sp.)生物量及代谢产物产量,研究了7种浓度的脱落酸(ABA)在三种氮素营养水平下对紫球藻生长的影响,结果表明,ABA浓度在0.01～1.0 mg·L-1范围内,外源ABA浓度的升高有利于紫球藻增殖,对紫球藻的干重、叶绿素a含量、蛋白质含量、胞外多糖含量、藻红素含量都有一定的促进作用;但在ABA浓度在10 mg·L-1时,则呈递抑制趋势.其中中氮水平下外源ABA浓度1.0 mg·L-1是紫球藻生长的最佳浓度,最有利于紫球藻生物量及代谢产物的积累,此时紫球藻的生物量为3.463 mg·mL-1,胞外多糖含量最高,占藻体干重的34.31%.在高氦营养水平下还能体现ABA对紫球藻生长所起的作用,而低氮下生长的紫球藻对ABA的作用不敏感.     

超声波降解紫球藻胞外多糖研究

通过测定溶液粘度,判断超声波降解紫球藻(Porphyridium cruentumi)胞外多糖的效果。运用均匀设计对超声波降解紫球藻胞外多糖的影响因素(处理振幅、时间、脉冲)进行优化,获得超声波处理的最佳条件:振幅39%、处理时间245s和脉冲9.5s。在最佳条件下,胞外多糖的粘度为2.98mm²/s,与预测值一致。采用DPS软件对实验结果进行二次多项式分析与拟合,并对模型和回归系数进行显著性检验,建立了以胞外多糖粘度为目标的回归方程式。     

半导体激光对紫球藻生物学效应的影响

采用波长650nm,功率40mW,功率密度13W／cm^2的半导体激光,对紫球藻（Porphyridium cruentum)进行诱变,辐照时间分别为5min、10min、20min。通过对紫球藻每天细胞数、叶绿素a、第7d细胞干重和胞外多糖产量的测定可知：与对照组比较,3个处理对紫球藻生长及胞外多糖产生影响,5min、10min均有不同程度的促进作用,其中5min作用最明显,而20min则有抑制作用。     

平板式光生物反应器中紫球藻培养条件的优化

研究了平板式光生物反应器中紫球藻(Porphyridium cruentumNaegeli)的培养条件,运用均匀设计法对光照强度、通气速率、装液量、接种密度以及pH等影响紫球藻生长的因素进行优化,获得了在平板式光生物反应器中培养紫球藻的最佳条件:光照强度10 000 lx、通气速率350 L.h-1、装液量6 L、藻细胞接种密度1.1×106mL-1、pH9.0。在最佳条件下藻体的生物量产率和生物量产量分别达到0.431 g.L-1.d-1和3.240 g.L-1,最大生长速率达0.652 g.L-1.d-1,胞外多糖含量高达0.665 g.L-1。另外,在培养过程中隔天补充培养液有利于紫球藻生物量的增加和胞外多糖的产生。     

倍频Nd∶YAG激光对紫球藻生长与胞外多糖产量的影响

紫球藻经Nd:YAG激光(波长1060 nm)照射,以不同时间设置处理剂量,比较紫球藻细胞活力、生长速率、生物量、胞外多糖产量等方面的变化.高剂量处理组(Y-20、Y-30和Y-40)致死率为17～44%,继代培养后生长迅速,K值分别比对照组提高18%、23.4%和15.3%.各处理组培养10天后收获的生物量与对照组无显著差别,但胞外多糖产量均有较大幅度提高,增幅达50～150%.此外,YAG激光照射对藻细胞叶绿素含量也有影响.YAG激光有望成为紫球藻优良藻株选育的较佳诱变剂.     

倍频Nd：YAG激光对紫球藻生长与胞外多糖产量的影响

紫球藻经Nd:YAG激光（波长1060nm)照射,以不同时间设置处理剂量,比较紫球藻细胞活力,生长速率,生物量,胞外多糖产量等方面的变化。高剂量处理组（Y-20、Y-30和Y-40）致死率为17-44%,继代培养后生长迅速,K值分别比对照组提高18%、23.4%和15.3%,各处理组培养10天后收获的生物量与对照组无显著差异,但胞外多糖产量均有较大幅度提高,增幅达50-150%。此外,YAG激光照射对藻细胞叶绿素含量也有影响,YAG激光有望成为紫球藻优良藻株选育的较佳诱变剂。     

紫外线对紫球藻的生物学效应

采用紫外线辐照紫球藻,处理剂量分别为10s、20s、30s、60s、90s、120s和180s。结果表明,10s剂量对紫球藻有促长作用,20s及30s对藻细胞生长和各项生理生化指标影响不显著,30s以上处理对其生长以及代谢产物的合成与分泌有抑制作用,且照射时间越长,抑制越强烈,紫球藻对紫外线的耐受极限为120s。结果还表明,高剂量(20~90s)照射能增加单细胞代谢产物含量;经紫外线辐照后的藻细胞第二代培养物在生长速度、叶绿素a含量、胞外多糖和β-胡萝卜素产量等与对照组相似。     

枸杞多糖与黄芪多糖抑菌活性的研究*

目的:研究黄芪多糖和枸杞多糖的抑菌活性并探讨不同pH值对其抑菌活性的影响。方法:采用滤纸片扩散法,分析不同浓度黄芪多糖和枸杞多糖在不同pH值下对几种常见细茵和霉菌(大肠杆菌、沙门氏菌、金黄色葡萄球菌、黑曲霉、产黄青霉)的抑制效果。结果:对于细菌,枸杞多糖8mg/mL时出现抑菌圈,而黄芪多糖0.02 mg/mL时效果最佳;对于霉菌,随着枸杞多糖浓度的增大,抑菌圈的直径增大,而黄芪多糖0.02 mg/mL时效果最佳;当枸杞多糖和黄芪多糖在pH6的条件下,二者抑菌活性均最强。结论:枸杞多糖和黄芪多糖对细菌、霉菌都有一定的抑制效果,pH值可影响枸杞多糖和黄芪多糖的抑菌效果。     

Extraction, purification and antibacterial activities of a polysaccharide from spent mushroom substrate

To contribute towards effective exploitation and utilization of spent mushroom substrate (SMS), a water-soluble polysaccharide named PL was isolated and purified from SMS. The total sugar content and monosaccharide composition were analyzed by phenol-sulfuric acid method and capillary electrophoresis, and infrared spectroscopy was also performed for structure characterization. The results showed that the total sugar content of crude polysaccharide from SMS was about 25.8%, the polysaccharide contained two fractions (PL1 and PL2), which was mainly composed of glucose, rhamnose and mannose with a molar ratio of 1:3.13:1.16. The attributions of the main absorptions of both PL1 and PL2 were characteristic of glycosidic structures, and the FT-IR spectra of PL2 and lentinan were very similar. Escherichia coli, Staphylococcus aureus and Sarcina lutea were used to study the antibacterial activity and minimal inhibitory concentrations (MICs) of the polysaccharide. The antibacterial activity of polysaccharide from SMS against E. coli was the strongest, while the weakest against S. lutea, and the MICs of PL2 were 12.5, 25 and 100 μg/mL, respectively.     

Sphagnan – a pectin-like polymer isolated from Sphagnum moss can inhibit the growth of some typical food spoilage and food poisoning bacteria by lowering the pH

Aims:  Investigate if the antibacterial effect of sphagnan, a pectin-like carbohydrate polymer extracted from Sphagnum moss, can be accounted for by its ability to lower the pH.
Methods and Results:  Antibacterial activity of sphagnan was assessed and compared to that of three other acids. Sphagnan in its acid form was able to inhibit growth of various food poisoning and spoilage bacteria on low-buffering solid growth medium, whereas sphagnan in its sodium form at neutral pH had no antibacterial activity. At similar acidic pH, sphagnan had comparable antibacterial activity to that of hydrochloric acid and a control rhamnogalacturonan pectin in its acid form.
Conclusions:  Sphagnan in its acid form is a weak macromolecular acid that can inhibit bacterial growth by lowering the pH of environments with a low buffering capacity.
Significance and Impact of the Study:  It has previously been suggested that sphagnan is an antimicrobial polysaccharide in the leaves of Sphagnum moss with a broad range of potential practical applications. Our results now show that sphagnan in its acid form can indeed inhibit bacterial growth, but only of acid-sensitive species. These findings represent increased knowledge towards our understanding on how sphagnan or Sphagnum moss might be used in practical applications.     

Synthesis and Study of Biological Activity of Sulfamic Polysaccharide Derivatives

New water-soluble derivatives of starch, pectin, and Na-CMC containing the sulfamic groups have been obtained by the reaction of sulfamic acid with dialdehyde polysaccharide derivatives. The structure and composition of the resulting compounds have been studied by IR spectroscopy, elemental (nitrogen and sulfur) analysis, and X-ray diffraction. The sulfamic derivatives of starch, pectin, and Na-CMC with a different content of the sulfamic groups have been obtained by varying the ratio of sulfamic acid to the dialdehyde polysaccharide derivatives. The optimal–СНО: NH₂SO₃H ratio was found to be 1: 2.5. The interaction rate of sulfamic acid with the dialdehyde derivatives of starch, pectin, and Na-CMC has been evaluated. The antibacterial and antifungal effects of sodium salts of the sulfamic starch, pectin, and Na-CMC derivatives against Gram-positive and Gram-negative bacteria and fungi have been studied at different concentrations (10, 25, 50 mg/mL) by the disk diffusion method. The synthesized compounds have not been found to exhibit antifungal activity against Candida albicans. Nevertheless, they have been shown to have the antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Proteus vulgaris, Streptococcus faecalis, Streptococcus pyogenes, and Streptococcus faecalis at the concentration of 50 mg/mL. The concentration dependence of antibacterial action of sodium salts of the starch, pectin, and Na-CMC sulfamic derivatives has been demonstrated. The antibacterial activity of the drugs has been found to directly depend on the content of the sulfamic groups in polysaccharides. The results on the acute toxicity of the sulfamic polysaccharide derivatives have shown that these compounds can be attributed to low-toxicity substances of Class V.     

Suppressor T cells regulating the cell-mediated immune response to Pseudomonas aeruginosa can be generated by immunization with anti-bacterial T cells

We previously demonstrated that immunization with low (10 micrograms) doses of high m.w. polysaccharide from the gram-negative bacterium Pseudomonas aeruginosa generates T cells that suppress the ability of antibacterial T cells (Tab) to protect against bacterial infection. The current studies indicate that Ts cells with properties identical to those elicited by low dose polysaccharide immunization can be generated by immunization with Tab. Tab-elicited Ts cells can abrogate in vivo induction and in vitro and in vivo expression of antibacterial T cell activity. Tab-elicited Ts are Ag-specific and H-2 restricted in their suppressor activity. Non-immune T cells fail to elicit suppressor activity. These studies provide additional evidence that the protective T cell response to P. aeruginosa is controlled by a network of T cells that are probably recognizing idiotypic determinants on P. aeruginosa-immune T and B cells.     

藿香枝叶多糖的提取工艺及其清除羟基自由基作用研究

采用苯酚-硫酸,并以超声法辅助提取野生藿香枝叶中的多糖,设计L9(34)正交试验考察料液比、pH和超声提取时间对藿香枝叶中多糖提取的影响,并对藿香多糖进行清除.OH试验。结果表明,藿香多糖的最佳提取工艺为料液比1∶10,超声提取30min,pH值为6,可使藿香多糖的提取率高达7.50%。藿香多糖对羟基自由基有显著地清除效果,表明其有抗氧化作用。     

明胶/纳米SiO2复合膜的体外抑菌试验

本实验测定了明胶、纳米SiO2、明胶/纳米SiO2复合膜对6种供试菌的抑菌效果,结果表明,明胶不具有抑菌作用;纳米SiO2对供试菌具有较好的抑茵效果;20种明胶/纳米SiO2复合膜材料中,有11种复合膜对供试菌具有较强的抑菌效果.用琼脂平板稀释法测定了具有较强抑菌作用复合膜的最低抑菌浓度(MIC),结果表明,最低抑菌浓度因膜的配比和菌种类型的不同而有所差异,为16.0 mg/mL～256.0 mg/mL.复合膜对蜡样芽孢杆菌的最低抑菌浓度(MIC)最大(256.0 mg/mL),而对其他供试菌相对较低,为64.0 mg/mL～128.0 mg/mL.     

金银花和苦丁茶多糖提取物抗菌活性研究

从金银花和苦丁茶中提取水溶性多糖,利用圆形纸片法对7种常见的食品腐败菌进行抗菌试验。结果表明:金银花和苦丁茶多糖提取物都具有一定程度的抗细菌与抗真菌活性,抗细菌活性比抗真菌活性强;苦丁茶多糖提取物对金黄色葡萄球菌的最低抑菌浓度为5mg/ml。     

纳米银/植物源复合抗菌剂的制备及其抑菌性能的研究

以硝酸银作为银源,水溶性淀粉作保护剂,丙酮酸钠作还原剂,氨水提供碱性环境来制备纳米银胶,并以聚乙烯吡咯烷酮(PVP)作分散稳定剂,复配红景天提取液和无患子提取液制备出纳米银/植物源复合抗菌剂。实验结果表明,纳米银胶或植物提取液仅对部分细菌或霉菌有较强抑制效果,而复合抗菌剂对细菌、霉菌均有很强抑制效果。在湿巾液中添加0.5%复合抗菌剂时,其对大肠杆菌,金黄色葡萄球菌和白色念珠菌的抗菌效率可达99%,且经过常温六个月、高温55℃一个月保存后,其抗菌活性分别可达到95%、90%左右,表明复合抗菌剂具有较强的抗菌效率及抗菌稳定性。     

塔拉种子多糖脱蛋白的正交优化及其抗氧化性研究

以塔拉（Caesalpinia spinosa）种子为原料,研究了塔拉种子多糖的脱蛋白工艺及塔拉多糖的抗氧化性质。以多糖损失率和蛋白脱除率为评价指标,比较Sevage法、三氯乙酸法和木瓜蛋白酶法对塔拉多糖的脱蛋白效果。利用正交优化组合实验设计原理,采用四因素三水平的正交分析法,对木瓜蛋白酶法脱蛋白进行正交优化。结果表明：塔拉多糖最佳脱蛋白工艺条件为酶添加量0.15mL、酶解时间90min、酶解温度60℃、酶解pH=6,蛋白脱除率95.19%,多糖保留率75.02%。通过对塔拉多糖抗氧化性的研究,发现塔拉多糖总抗氧化性较好,对DPPH自由基有较强的清除作用。