竹荪提取物对食源性细菌的抑菌特性研究

利用4种方法获得棘托竹荪和长裙竹荪的提取物,以5种食源性致病细菌单增李斯特菌、金黄色葡萄球菌、肠炎沙门菌、大肠埃希菌O157H7、副溶血性弧菌为供试菌,比较其MIC(最低抑菌浓度)和MBC(最低杀菌浓度)。超声波辅助破碎竹荪干粉,制取水、乙醇和石油醚为提取介质的浸提物以及竹荪挥发油,利用液体培养基连续稀释法对5种供试菌进行抑菌实验。所有提取物对供试菌都具有抑菌活性;长裙竹荪提取物的综合抑菌能力高于棘托竹荪提取物;水提物和挥发油的抑菌效果最好;2种竹荪提取物对革兰阴性菌和革兰阳性菌的抑菌作用存在互补效应。竹荪提取物对5种食源性细菌的生长有较好的抑制作用,可应用2种竹荪的提取物复配,开发天然食品防腐剂。     

长裙竹荪正己烷提取物化学组成及抑菌活性研究

以正己烷为溶剂,对长裙竹荪子实体进行索氏提取,提取率为1.36%。应用GC-MS对提取物的化学成分进行分析,Rxi-1ms柱分离,质谱解析鉴定出55种成分,其中23种成分是首次从竹荪属中检测出来,其主要成分为:羧酸、醇、酮、倍半萜、芳香烃、酯等。提取物对伤寒杆菌、金黄色葡萄球菌、变形杆菌和枯草芽孢杆菌有很好的抑制作用。     

长裙竹荪抑菌活性物质的分离纯化及其抑菌效果

长裙竹荪Dictyophora indusiata是珍贵的食药用真菌,具有很强的抑菌作用,在天然防腐剂开发方面具有广阔的应用前景。本研究以长裙竹荪的抑菌活性为指标,通过萃取、3次不同流动相的硅胶柱层析、1次反相柱层析和薄层层析法对竹荪提取物进行分离纯化,得到一个抗菌活性强的单体化合物。根据核磁共振波谱等数据分析,推断该化合物为间苯三酚。以巨大芽孢杆菌和肠炎沙门氏菌为供试菌,用平板打孔法及原位抑菌法测定该化合物的抑菌效果,结果表明：该化合物对这两种菌有很强的抑制作用,半抑制浓度分别为83.06μg/mL和51.58μg/mL。本研究首次从长裙竹荪中获得具有抗菌活性的单体化合物间苯三酚,为竹荪天然抗菌物质的开发提供理论依据。     

Biolog-ECO解析长裙竹荪栽培种覆土层土壤微生物群落功能多样性特征

为了解不同基质培育长裙竹荪栽培种覆土层土壤微生物群落功能多样性特点,运用Biolog-ECO技术,分析了以生橡胶木屑和熟橡胶木屑为基质的覆土层土壤微生物群落功能多样性。结果表明:覆土层土壤微生物平均吸光值(AWCD)总体变化趋势和对六大类碳源的利用情况均为生料组大于熟料组和对照组,且熟料组和对照组基本一致;生料组Mc Intosh指数与熟料组、对照组间显著差异,但3组之间的Shannon指数(H')、Simpson指数(D)和Pielou指数(E)的差异均不显著。熟料作为培育长裙竹荪栽培种的基质加入,既增强了覆土层土壤的通透性,又为长裙竹荪菌丝的生长繁殖提供了丰富的营养和良好的土壤微环境,更有利于长裙竹荪栽培种菌丝的生长和繁殖,对长裙竹荪栽培种的培育质量具有重要的作用。     

鬼笔属3种真菌的个体发育及形态观察

为揭示鬼笔属真菌子实体各器官的发育过程和形态特点,采用石蜡切片法对驯化栽培出菇的超短裙竹荪Phallus ultraduplicatus、冬荪P. dongsun和红托竹荪P. rubrovolvatus不同发育阶段的菇蕾进行切片显微观察,结合菌丝、孢子和子实体形态比较3个种的个体发育及形态特点。结果表明：3个种的菌丝和孢子形态差异不明显,超短裙竹荪子实体的菌裙质地较薄、长约4 cm、带有不规则形状的网孔;红托竹荪的菌裙质地较厚、长约7 cm、带有圆形网孔;冬荪仅有1层薄菌幕。3个种的菇蕾内部发育顺序一致,最先发育的都是胶质腔,接着是子实层、菌柄腔、菌盖、菌柄、菌裙（菌幕）;发育过程中菌丝形态经历从疏松到紧密、从交织状到组织状的变化过程;各器官发育完成的时间不同,在菇蕾直径相同的条件下,冬荪发育最快,在菇蕾直径为2 cm时,各器官基本发育完成,红托竹荪在菇蕾直径为3 cm时发育完成,超短裙竹荪的菇蕾发育最慢,在直径达到4 cm时各器官才能发育完成。为鬼笔类真菌的分类研究和亲缘关系探索提供了依据。     

红托竹荪菌托多糖的提取及抗肿瘤活性的初步研究

红托竹荪菌托经热水提取、酒精沉淀、脱蛋白后的粗多糖,其得率远大于其菌丝体和子实体的其他部位及香菇子实体。菌托粗多糖进一步用DEAE纤维素柱和Sephadex G-75分离纯化,得到两个组分DRVP1与DRVP2,对分离得到的主要多糖通过高效液相色谱（HPLC）、红外光谱（IR）等进行结构分析。DRVP1的相对分子质量（Mr）为1.47×104,红外光谱数据显示为β-型甘露糖苷。体外试验表明,红托竹荪菌托多糖的组分DRVP1对小鼠S180肉瘤有一定的抑制作用。     

红托竹荪菌托多糖的提取及抗肿瘤活性的初步研究

红托竹荪菌托经热水提取、酒精沉淀、脱蛋白后的粗多糖,其得率远大于其菌丝体和子实体的其他部位及香菇子实体。菌托粗多糖进一步用DEAE纤维素柱和Sephadex G-75分离纯化,得到两个组分DRVP1与DRVP2,对分离得到的主要多糖通过高效液相色谱（HPLC）、红外光谱（IR）等进行结构分析。DRVP1的相对分子质量（Mr）为1．47x10^4,红外光谱数据显示为β-型甘露糖苷。体外试验表明,红托竹荪菌托多糖的组分DRVP1对小鼠S180肉瘤有一定的抑制作用。     

竹荪的菌丝培养及其抗菌性的初步研究

对长裙竹荪（Ｄｉｃｔｙｏｐｈｏｒａ　ｉｎｄｕｓｉａｔａ）等３种的菌丝培养进行了研究并初步测定菌丝的抗菌谱。培养液的 ｐＨ和液体培养基粘度是竹荪在液体中生长的关键因素,在 ｐＨ４.７和添加 ０.５％ＣＭＣ－Ｎａ的合适培养条件下竹荪菌丝在液体中能够生长,菌丝得率达到１％。还探讨了其他几种竹荪菌丝的培养方法,为竹荪菌丝的大量生产提供了依据。另外,在竹荪抗菌性实验中发现,竹荪对许多易造成食品腐败的细菌和酵母具有抗性,但没有发现其对霉菌有抗性。     

不同产地松茸氨基酸与矿质元素的比较研究

以西南地区的三种松茸子实体为材料 ,用柱前衍生高效液相色谱法和原子吸收分光光度法分别对松茸子实体的氨基酸和矿质元素进行了测定 ,氨基酸的测定结果表明 :采自四川通江的松茸子实体的氨基酸总含量最高 ,为 10 .5 82 % ;其次为采自重庆的松茸子实体为 9.6 0 7% ;最低的为采自云南昌隆的松茸子实体 ,为 3.381%。矿质元素的测定结果表明 :所测定的七种元素中 ,含量较高的有三种 ,分别为K ,Ca和Mg ,平均含量分别为 3.1× 10 4mg/Kg ,1.6 2× 10 3mg/Kg和 5 .5 8× 10 2mg/Kg。在这三种松茸子实体中云南昌隆的Fe ,Mn含量最高 ,分别为 6 2 .2 82mg/Kg和 6 .0 6 4mg/Kg ;重庆的Cu含量最高 ,为 5 2 .84 5mg/Kg;Zn的含量在这三种子实体中差异不大。     

红托竹荪菌托多糖的提取及抗肿瘤活性的初步研究

红托竹荪菌托经热水提取、酒精沉淀、脱蛋白后的粗多糖,其得率远大于其菌丝体和子实体的其他部位及香菇子实体.菌托粗多糖进一步用DEAE纤维素柱和Sephadex G-75分离纯化,得到两个组分DRVP1与DRVP2,对分离得到的主要多糖通过高效液相色谱(HPLC)、红外光谱(IR)等进行结构分析.DRVP1的相对分子质量(Mr)为1.47×104,红外光谱数据显示为β-型甘露糖苷.体外试验表明,红托竹荪菌托多糖的组分DRVP1对小鼠S180肉瘤有一定的抑制作用.     

虎纹捕鸟蛛凝集素-I(SHL-I)的细胞凝集活性分析

采用人红细胞悬液在微量血凝板上测试了 1 1种单糖对虎纹捕鸟蛛凝集素 - I ( Selenocosmiahuwena lectin- I,SHL - I)的凝集活性的影响 .测试的糖类包括 D-半乳糖 ,甲基 -α- D甘露糖苷 ,D-甘露糖 ,D-甘露糖胺 ,D-葡萄糖 ,N-乙酰 - D-葡萄糖胺 ,D-葡萄糖胺 ,L -木糖 ,L-岩藻糖 ,N-乙酰神经氨酸 ,N-乙酰 - D-半乳糖胺 .测试结果表明 ,只有 D-甘露糖胺对 SHL- I的凝集活性表现出明显的抑制作用 .说明 D-甘露糖胺可能是与 SHL- I专一性结合的糖基 .温度适应性实验表明 SHL- I有较高的热稳定性 ,经 1 0 0℃处理 30 min,仍保持大部分凝集活性 .p H适应性实验表明 ,在碱性 p H环境下SHL- I的凝集活性明显下降 .     

Getting the Jump on the Development of Bullfrog Oil Microemulsions: a Nanocarrier for Amphotericin B Intended for Antifungal Treatment

Amphotericin B (AmB), a potent antifungal drug, presents physicochemical characteristics that impair the development of suitable dosage forms. In order to overcome the AmB insolubility, several lipid carriers such as microemulsions have been developed. In this context, the bullfrog oil stands out as an eligible oily phase component, since its cholesterol composition may favor the AmB incorporation. Thus, the aim of this study was to develop a microemulsion based on bullfrog oil containing AmB. Moreover, its thermal stability, antifungal activity, and cytotoxicity in vitro were evaluated. The microemulsion formulation was produced using the pseudo-ternary phase diagram (PTPD) approach and the AmB was incorporated based on the pH variation technique. The antifungal activity was evaluated by determination of minimal inhibitory concentration (MIC) against different species of Candida spp. and Trichosporon asahii. The bullfrog oil microemulsion, stabilized with 16.8% of a surfactant blend, presented an average droplet size of 26.50?±?0.14 nm and a polydispersity index of 0.167?±?0.006. This system was able to entrap AmB up to 2 mg mL^?1. The use of bullfrog oil as oily phase allowed an improvement of the thermal stability of the system. The MIC assay results revealed a growth inhibition for different strains of Candida spp. and were able to enhance the activity of AmB against T. asahii. The microemulsion was also able to reduce the AmB toxicity. Finally, the developed microemulsion showed to be a suitable system to incorporate AmB, improving the system’s thermal stability, increasing the antifungal activity, and reducing the toxicity of this drug.     

双歧啤酒对常见病原菌的拮抗作用

目的了解双歧啤酒对常见病原菌的拮抗效果及影响因素。方法采用纸片扩散法（K-B法）、琼脂打孔法和试管稀释法（MIC法）测定双歧啤酒及其处理物（加热、不同pH）对常见病原菌的抑菌作用,同时用某市售啤酒作为对照。结果双歧啤酒及其处理物均具有一定的抑菌活性。结论双歧啤酒对常见病原菌有一定的抑菌作用,抑菌活性强于市售啤酒。     

双歧杆菌发酵果蔬汁的体外生物拮抗作用研究

目的研究双歧杆菌发酵果蔬汁的体外生物拮抗作用及影响因素。方法采用纸片扩散法(K-B法)、琼脂打孔法和试管稀释法(MIC)研究双歧杆菌发酵果蔬汁对常见病原菌的体外拮抗作用,并讨论pH、加热对拮抗作用的影响。结果发酵果蔬汁及其处理物(加热、不同pH)均具有一定的抑菌作用。结论双歧杆菌发酵果蔬汁对常见病原菌有一定的拮抗作用。     

Salt-induced stabilization of apoflavodoxin at neutral pH is mediated through cation-specific effects

Electrostatic contributions to the conformational stability of apoflavodoxin were studied by measurement of the proton and salt-linked stability of this highly acidic protein with urea and temperature denaturation. Structure-based calculations of electrostatic Gibbs free energy were performed in parallel over a range of pH values and salt concentrations with an empirical continuum method. The stability of apoflavodoxin was higher near the isoelectric point (pH 4) than at neutral pH. This behavior was captured quantitatively by the structure-based calculations. In addition, the calculations showed that increasing salt concentration in the range of 0 to 500 mM stabilized the protein, which was confirmed experimentally. The effects of salts on stability were strongly dependent on cationic species: K(+), Na(+), Ca(2+), and Mg(2+) exerted similar effects, much different from the effect measured in the presence of the bulky choline cation. Thus cations bind weakly to the negatively charged surface of apoflavodoxin. The similar magnitude of the effects exerted by different cations indicates that their hydration shells are not disrupted significantly by interactions with the protein. Site-directed mutagenesis of selected residues and the analysis of truncation variants indicate that cation binding is not site-specific and that the cation-binding regions are located in the central region of the protein sequence. Three-state analysis of the thermal denaturation indicates that the equilibrium intermediate populated during thermal unfolding is competent to bind cations. The unusual increase in the stability of apoflavodoxin at neutral pH affected by salts is likely to be a common property among highly acidic proteins.     

新型α-半乳糖苷酶的稳定性研究

目的：观察脆弱类杆菌来源的α-半乳糖苷酶（GAL）在不同pH缓冲液、不同温度下的稳定性,以及不同离子及还原剂对酶活性的影响。方法：以GAL对单糖底物对硝基-苯基-α-D-吡喃半乳糖苷（PNPG）的活性为主要检测指标,观察不同离子及还原剂等对酶活性的影响;观察GAL在不同pH缓冲液中和不同温度下的稳定性。结果：钙离子、锌离子、钴离子和高浓度的锰离子增强酶的活性,DTT抑制酶的活性,螯合剂EDTA的加入提高了酶活性。GAL在pH4.6～7.5时保存1 h后稳定性很好,能保持最高活性的90％以上;在4℃～45℃下保存的稳定性最好,45℃开始活性下降。结论：GAL具有很好的温度稳定性和pH稳定性,使其适用于血型转变和异种移植。     

Streptomyces halstedii K122 produces the antifungal compounds bafilomycin B1 and C1

Streptomyces halstedii K122 was previously found to produce antifungal compounds on solid substrates that inhibit radial growth of fungi among Ascomycetes, Basidiomycetes, Deuteromycetes, Oomycetes, and Zygomycetes, and strongly affected hyphal branching and morphology. During growth of S. halstedii K122 in submerged culture, no antifungal activity could be detected. However, cultivation of S. halstedii in thin (1 mm) liquid substrate layers in large surface-area tissue culture flasks caused intense growth and sporulation of S. halstedii K122, and the biologically active compounds could be extracted from the mycelium with methanol. Antifungal compounds were purified using C18 solid phase extraction and silica gel column chromatography, and identified as bafilomycins B1 and C1, using 2D NMR and FAB MS. Production of bafilomycins, which are specific inhibitors of vacuolar ATPases, has not been reported from S. halstedii previously. Minimum inhibitory concentrations (MIC) of bafilomycins B1 and C1, amphotericin B, and nikkomycin Z were determined at pH 5.5 and 7.0 for the target fungi Aspergillus fumigatus, Mucor hiemalis, Penicillium roqueforti, and Paecilomyces variotii. Penicillium roqueforti was the most sensitive species to all the compounds investigated. The MIC values for amphotericin B were 0.5-4 micrograms.mL-1 for the fungi tested, and pH did not affect the toxicity. The MIC values for nikkomycin Z ranged from < 0.5 microgram.mL-1 for Mucor hiemalis to > 500 micrograms.mL-1 for Aspergillus fumigatus, and pH had no influence on toxicity. Bafilomycins B1 and C1 were equally active against the fungal species tested, with MIC values in the range of < 0.5-64 micrograms.mL-1. All fungi were more sensitive to both bafilomycin B1 and C1 at pH 7.0 than at pH 5.5.     

Lysosomotropic N,N- dimethyl alpha-aminoacid N-alkyl esters and their quaternary ammonium salts as plasma membrane and mitochondrial ATPases inhibitors

A set of n-alkyl esters of N,N-dimethylglycine (DMG-n) and their methobromides (DMGM-n) was synthesized, and their activities on yeast Saccharomyces cerevisiae were compared. The compounds differ in the number of carbon atoms in the aliphatic chain. Aminoesters with 12 carbon atoms appeared to be most active. Unlike quaternary ammonium salts previously tested, the activities of the compounds were not pH-dependent; the minimal inhibitory concentrations (MIC) were identical at pH 8 and at pH 6. In contrast to quaternary ammonium salts, aminoesters showed similar effects on respiratory sufficient (rho+) and respiratory deficient (rho0) mutants. When tested on glucose stimulated proton extrusion, aminoesters applied at MIC increased external pH. Aminoesters inhibited the plasma membrane H+-ATPase, whereas they were less inhibitory on the mitochondrial ATPase. In order to further compare the aminoesters and their corresponding quaternary ammonium salts, derivatives of N,N-dimethylalanine (DMAL-n and DMALM-n, respectively) were synthesized. The quaternary ammonium salts appeared to have a higher inhibitory potency than aminoesters, especially at pH 8, and alanine derivatives inhibited growth at a lower concentration than glycine derivatives. Both alanine derivatives of the aminoester and the quaternary ammonium salt inhibited the plasma membrane H+- ATPase at lower concentrations than glycine derivatives, but the alanine aminoester was without a detectable effect on the mitochondrial ATPase.     

贯叶连翘总提取物对致病细菌的抗菌作用

用平板抑菌法检测了贯叶连翘总提取物对细菌的抗菌谱范围,用最低抑菌浓度(MIC)检测法、最低杀菌浓度(MBC)检测法检测了其抗菌作用的强弱。结果表明提取物对供试革兰氏阳性菌菌株均有较强的抑菌和杀菌作用,对极少数革兰氏阴性菌菌株有较弱的抑菌作用,无杀菌作用。该结果提示提取物抗菌作用可能与细菌细胞壁的结构与组成相关。     

四种中药对耐药肺炎链球菌的抑菌作用

目的探讨中草药对肺炎链球菌的体外抑菌作用。方法用临床分离细菌为受试菌,采用琼脂稀释法,观察黄连、黄岑、黄柏和连翘等4种中草药水煎剂的最低抑菌浓度。结果4种中草药水煎剂均有不同程度的抑菌作用,其中黄连抑菌效果最好,黄连水煎剂对肺炎链球菌M IC范围达7.8-15.63 g/L,M IC50为8 g/L,M IC90为8 g/L。黄岑、黄柏也有较好的抑菌效果,连翘抑菌效果较弱。结论黄连等中药在体外对肺炎链球菌均有良好抑菌作用。