干酪乳杆菌JH-23中单胺氧化酶抑制剂的初步分离

采用丙酮抽提法得到了干酪乳杆菌JH-23中具有抑制单胺氧化酶作用的无细胞提取物,并利用硅胶薄层色谱从中分离出4组活性片段,其中片段Ⅰ对单胺氧化酶的抑制效果最佳,反应浓度在120μg/mL时抑制率为55.4%。该活性片段经国家新药筛选中心确证有抑制单胺氧化酶的效果,具有潜在的抗衰老作用。     

一种白僵菌中MAO抑制剂的分离纯化和结构鉴定

本研究对前期筛选出的一株具有较强的单胺氧化酶(MAO)抑制活性的白僵菌菌株Ba02进行了液体培养;通过不同提取剂的提取效果比较,发现乙酸乙酯能较好地提取出该发酵液中单胺氧化酶抑制剂。通过活性指导下的色谱分离,从乙酸乙酯提取物中得到了一种深红色粉末状化合物。活性测定结果显示该化合物在15μg.mL-1时对MAO-A和MAO-B的抑制率分别为97.50%和95.34%。MS和NMR的鉴定结果表明该化合物为卵孢菌素(Oosporein)。虽然该化合物是一已知化合物,但其对单胺氧化酶的抑制活性尚属首次发现。     

泡菜、传统腊肠中降胆固醇乳酸菌的筛选及鉴定

摘要：【目的】筛选具有降胆固醇功能的乳酸菌菌株,为乳酸菌体外、体内的降胆固醇生理特性和机理研究奠定基础。【方法】以碳酸钙-MRS选择性培养基(Calcium Carbonate-Man Rogosa and Sharp Medium)从中国传统食品泡菜、腊肠中筛选乳酸菌,应用改良的胆固醇筛选培养基筛选具有较高降胆固醇能力的乳酸菌菌株,并研究其耐酸性,耐胆盐活性,生长曲线及产酸特性;结合菌落形态学、接触酶反应、革兰氏染色、碳水化合物微量鉴定管及16SrRNA寡核苷酸碱基序列分析鉴定菌株。【结果】筛选得到的两     

不同菌株白僵菌代谢产物对大鼠肝线粒体单胺氧化酶的抑制作用研究

对8株白僵菌[Beauveriasp.]代谢产物的抑制单胺氧化酶的活性进行测定,发现菌株Ba02和Ba05的乙酸乙酯提取物在终浓度为55μg/mL时,对单胺氧化酶A型(MAO＿＿A)有较强的抑制活性,其抑制率分别为95.9%和83.4%;Ba02及Ba03菌株脂溶性成分在终浓度为55μg/mL时对单胺氧化酶B型(MAO＿B)有较强的抑制活性,其抑制率分别为94.8%和84.1%。浓度和抑制活性关系研究表明在一定浓度范围内Ba02菌株脂溶性成分对MAO的抑制活性呈量效关系,经计算得出其对MAO＿A、MAO＿B抑制的IC50分别为18.3μg/mL、28.2μg/mL;抑制特征曲线表明Ba02菌株脂溶性成分对MAO＿A呈竞争型抑制,对MAO＿B为混合型抑制,Km值分别为0.47×10-5mol/L,0.11×10-5mol/L。     

干酪乳杆菌JH23代谢产物对单胺氧化酶的抑制作用

本文旨在分离干酪乳杆菌JH-23中具有抑制单胺氧化酶(MAO)活性的代谢产物.用乙酸乙酯对干酪乳杆菌JH23发酵液进行抽提,得到的脂溶性成分和水溶性成分在一定浓度范围内对MAO的抑制活性呈较好的量效关系.其中,脂溶性成分对MAO-A、MAO-B抑制作用的IC50分别为1.97和5.67 mg/mL;水溶性成分对MAO-A、MAO-B抑制作用的IC50分别为1.59和4.83 mg/mL.抑制特征曲线显示,两者对MAO-A与MAO-B的抑制作用均呈竞争性抑制.从水溶性成分中分离得到的活性组分W1对MAO-A、MAO-B抑制作用的IC50分别为0.33和1.13 mg/mL.通过进一步分离,以及红外和质谱分析,确定了组分W1中具有较强MAO抑制作用的物质为琥珀酸.     

西藏林芝土壤放线菌抗肿瘤活性菌的筛选及鉴定

【目的】筛选并鉴定西藏林芝八一镇土壤中产生抗肿瘤活性物质的放线菌。【方法】用平板稀释法分离放线菌, 用MTT法和纸碟法对放线菌发酵产物进行体外抗肿瘤与抑菌活性检测, 并用多相分类技术对抗肿瘤活性菌株进行鉴定。【结果】共分离出29株放线菌, 得到6个抑制体外肿瘤细胞增殖的活性菌株(20.7%), 同时它们也具有抑菌活性, 其中4株菌的检测样品对Hela细胞的抑制率达80%以上。多相分类研究表明, 6个活性菌株分别隶属于链霉菌属(Streptomyces)的3个已知物种的变种。【结论】林芝八一镇土壤放线菌中蕴藏抗肿瘤活性链霉菌, 是一个潜在的抗肿瘤药用微生物资源。     

具有抑制单胺氧化酶作用的干酪乳杆菌JH-23的药敏性研究

采取纸片琼脂扩散法对具有抑制单胺氧化酶作用的干酪乳杆菌JH-23进行抗生素的敏感性测定.考察了不同菌液浓度、培养时间和接种方式对药敏性实验的影响,在此基础上建立了其对12类30种抗生素的药敏谱,以进一步全面评估该益生菌的安全性.     

一株产右旋糖苷酶海洋细菌的筛选鉴定

【目的】筛选鉴定产右旋糖苷酶的海洋细菌,并对其所产右旋糖苷酶的酶学性质及在变异链球菌牙菌斑生物膜中的应用进行初步研究。【方法】利用平板透明圈法从海洋环境中筛选产右旋糖苷酶的细菌,根据菌株形态特征、生理特征及16S rDNA序列确定其分类学地位,采用体外生物膜模型研究该酶对变异链球菌牙菌斑生物膜形成的抑制作用。【结果】从海泥中筛选出一株产右旋糖苷酶的细菌KQ11,初步鉴定为节杆菌(Arthrobacter sp.)。该菌株的最适生长温度为30°C,最适生长pH 7.5,最适生长NaCl浓度为0.4%。右旋糖苷酶的最适作用温度为45°C,最适作用pH为5.5。该酶能有效地抑制变异链球菌牙菌斑生物膜的形成。【结论】菌株KQ11右旋糖苷酶能够抑制变异链球菌牙菌斑生物膜的形成,可望用于漱口液等口腔护理产品中。     

五株乳酸菌和三株芽孢杆菌的生物学特性和功能

【背景】乳酸菌和芽孢杆菌是应用于生产最多的益生菌,但不同菌株间的生长特性均不相同,因此了解菌株的生物学特性具有重要意义。【目的】研究菌株的生物学特性,能合理地开发和利用菌株,以保证菌株生产应用的安全性。【方法】活化后鉴定5株乳酸菌和3株芽孢杆菌并对其形态进行观察,探究菌株的生长曲线、产酸能力及最适生长条件,测定菌株的抑菌活性和产酶性能,同时探究菌株的益生性和安全性。【结果】五株乳酸菌分别编号鉴定为干酪乳杆菌R1、副干酪乳杆菌R2、香肠乳杆菌R3、福莱乳杆菌R4和唾液乳杆菌R5;3株芽孢杆菌分别编号命名为贝莱斯芽孢杆菌Y1、枯草芽孢杆菌Y2和地衣芽孢杆菌Y3。八株菌形态结构均不相同但都为杆状,均在2–10 h为对数生长期,18–24 h为稳定期,培养24 h时乳酸菌和芽孢杆菌的活菌数均保持在10⁹和10⁸ CFU/mL,最适生长温度为37.0℃。乳酸菌具有较强的产酸能力和抑菌活性,芽孢杆菌有较强的产酶性,在人工胃液中都有较强的耐受性。八株菌都无溶血活性、无毒力基因、对抗生素都保持中度敏感以上;其中唾液乳杆菌有四环素耐药基因,但对四环素抗性为中度敏感。【结论】八株菌生长繁殖速度快,乳酸菌产酸能力和抑菌活性较强,芽孢杆菌具有较强的产酶性能,在体外具有较好的益生性和安全性,可应用于生产实践。     

产漆酶菌株筛选及一株产酶菌株的优化与鉴定

【目的】从26株真菌菌株中筛选高产漆酶菌株。【方法】采用愈创木酚法进行产漆酶菌株的筛选,通过正交实验对筛选出的高产菌株进行优化,并通过形态学和分子系统学对菌株进行鉴定。【结果】26株真菌菌株中有4株可产生漆酶,其中菌株H52.1为产漆酶最好菌株;菌株H52.1产漆酶优化培养基碳源为可溶性淀粉,氮源为硝酸铵,pH为8,金属离子为Ca2+;经鉴定,该菌株为大孢戴氏霉。【结论】大孢戴氏霉在产漆酶方面值得进一步研究开发。     

In vitro and in vivo inhibition of Helicobacter pylori by Lactobacillus casei strain Shirota

We studied the potential inhibitory effect of Lactobacillus casei strain Shirota (from the fermented milk product Yakult [Yakult Ltd., Tokyo, Japan]) on Helicobacter pylori by using (i) in vitro inhibition assays with H. pylori SS1 (Sydney strain 1) and nine H. pylori clinical isolates and (ii) the in vivo H. pylori SS1 mouse model of infection over a period of 9 months. In vitro activity against H. pylori SS1 and all of the clinical isolates was observed in the presence of viable L. casei strain Shirota cells but not in the cell-free culture supernatant, although there was profound inhibition of urease activity. In vivo experiments were performed by oral administration of L. casei strain Shirota in the water supply over a period of 9 months to 6-week-old C57BL/6 mice previously infected with H. pylori SS1 (study group; n = 25). Appropriate control groups of H. pylori-infected but untreated animals (n = 25) and uninfected animals given L. casei strain Shirota (n = 25) also were included in the study. H. pylori colonization and development of gastritis were assessed at 1, 2, 3, 6, and 9 months postinfection. A significant reduction in the levels of H. pylori colonization was observed in the antrum and body mucosa in vivo in the lactobacillus-treated study group, as assessed by viable cultures, compared to the levels in the H. pylori-infected control group. This reduction was accompanied by a significant decline in the associated chronic and active gastric mucosal inflammation observed at each time point throughout the observation period. A trend toward a decrease in the anti-H. pylori immunoglobulin G response was measured in the serum of the animals treated with lactobacillus, although this decrease was not significant.     

Evidence that gingko biloba extract does not inhibit MAO A and B in living human brain

Extracts of Ginkgo biloba have been reported to reversibly inhibit both monoamine oxidase (MAO) A and B in rat brain in vitro leading to speculation that MAO inhibition may contribute to some of its central nervous system effects. Here we have used positron emission tomography (PET) to measure the effects of Ginkgo biloba on human brain MAO A and B in 10 subjects treated for 1 month with 120 mg/day of the Ginkgo biloba extract EGb 761, using [11C]clorgyline and [11C]L-deprenyl-D2 to measure MAO A and B respectively. A three-compartment model was used to calculate the plasma to brain transfer constant K1 which is related to blood flow, and lambdak3, a model term which is a function of the concentration of catalytically active MAO molecules. Ginkgo biloba administration did not produce significant changes in brain MAO A or MAO B suggesting that mechanisms other than MAO inhibition need to be considered as mediating some of its CNS effects.     

Inhibition of rat brainstem monoamine oxidase activity by CGP 6085 A

CGP 6085 A [4-(5,6-dimethyl-2-benzofuranyl)piperidine] HCl, a known serotonin inhibitor, also inhibits rat brainstem monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B) in both in vivo and in vitro experiments. Serotonin (5-HT) deamination by MAO-A is inhibited 35% at a dose of 100 mg/kg i.p. in vivo. Similar experiments show a maximal 20% decrease in phenylethylamine (PEA) deamination by MAO-B at a dosage of 30 mg/kg i.p. Over the range of 0.1 to 10 mg/kg i.p., CGP 6085 A decreases 5-HIAA levels in the brainstem. This in vivo inhibition of MAO activity is confirmed by in vitro experiments. In vitro studies in rat brainstem mitochondrial preparations show a dose-dependent, reversible, inhibition of MAO using tyramine as the substrate for the enzyme reaction. With an in vitro IC50 of 2-3 microM, the potency of CGP 6085 A is comparable to pargyline.     

Irreversible inhibition of monoamine oxidase by some components of cigarette smoke

Inhibitory activity towards monoamine oxidase has been found in a solution of cigarette smoke. The inhibition was irreversible. When tissue slices of rat lung were incubated in the cigarette smoke solution or alternatively, exposed directly to cigarette smoke, monoamine oxidase activities were reduced drastically. Similarly, human saliva after cigarette smoking also exhibits considerable MAO inhibitory activity. When the amine substrates p-tyramine, serotonin and beta-phenylethylamine were incubated with the cigarette smoke solution, lipophilic adducts were formed non-enzymatically. The irreversible inhibition of MAO by cigarette smoke may well be related to the low platelet MAO associated with cigarette smokers as previously reported. The implication of such cigarette smoke-caused reduction of MAO activity in relation to Parkinsonism is discussed.     

Effects of Pesticides on Nitrite Oxidation by Nitrobacter agilis

The influence of pesticides on the growth of Nitrobacter agilis in aerated cultures and on the respiration of N. agilis cell suspensions and cell-free extracts was studied. Two pesticides, aldrin and simazine, were not inhibitory to growth of Nitrobacter, but five compounds [isopropyl N-(3-chlorophenyl) carbamate (CIPC), chlordane, 1,1-dichloro-2,2-bis (p-chlorophenyl) ethane (DDD), heptachlor, and lindane] prevented growth when added to the medium at a concentration of 10 mug/ml. Whereas CIPC and eptam prevented nitrite oxidation by cell suspensions, the addition of DDD and lindane resulted in only partial inhibition of the oxidation. Heptachlor and chlordane also caused only partial inhibition of oxidation, but were more toxic with cell-free extract nitrite oxidase. None of the pesticides inhibited the nitrate reductase activity of cell-free extracts, but most caused some repression of cytochrome c oxidase activity. Heptachlor was the most deleterious compound.     

[3H]HARMALINE AS A SPECIFIC LIGAND OF MAO A—I. PROPERTIES OF THE ACTIVE SITE OF MAO A FROM RAT AND BOVINE BRAINS

A high-affinity (K_d= 5.9 nM) specific binding site for [³H]harmaline was detected in membranes from rat and bovine brains. Studies of the regional and subcellular distributions of this binding indicated its close association with monoamine oxidase type A activity (MAO A) measured with [³H]serotonin ([³H]5-HT) as the substrate. Maximal binding capacity and MAO A activity were found in mitochondrial enriched fractions. Mitochondria of synaptosomal or extra-synaptosomal origin exhibited very similar properties with respect to [³H]harmaline binding characteristics and MAO A activity. Among psychoactive drugs, only monoamine oxidase inhibitors (MAO I) prevented the specific binding of [₃H]harmaline. Logit-log inhibition curves of binding by MAO I gave only one slope which was not significantly different from 1.0, suggesting the existence of only 1 category of specific sites for [³H]harmaline in the membrane preparations from rat and bovine brains. Consistent with the preferential inhibition of MAO A by harmaline, other MAO I of this class, i.e. clorgyline and Lilly 51641, were 10²-2 × 10³ times more efficient than deprenyl and pargyline, two inhibitors of MAO type B, in displacing [³H]harmaline from its specific binding site. K_i and IC₅₀ values for the inhibition of [³H]harmaline binding by MAO I and MAO substrates (tryptamine, 5-HT, norepinephrine) were almost identical with those characterizing their action on MAO A activity with [³H]5-HT as the substrate. In conclusion, the specific binding site for [³H]harmaline exhibited all the expected properties of the active site of MAO A. Like the technique of precipitation with a specific antibody, binding of [³H]harmaline should be of great help for studying the structural characteristics of the active site of MAO A and determining the number of MAO molecules in tissues under various physiological conditions.     

Proteolytic activity of lactobacilli in a model goats' milk curd system

Mesophyllic lactobacilli cultures propagated in MRS broth were inoculated in goats' milk curd slurries and incubated at 30C for 10 d. The micro-organisms tested were Lactobacillus casei subsp. casei IFPL 731 and IFPL 99, and Lactobacillus plantarum IFPL 3. Whole cells, cell-free extracts and cell lysates were evaluated for acceleration of proteolysis in the curd slurries. Conversion of water-soluble nitrogen to non-protein nitrogen and amino acid nitrogen, reverse phase-HPLC peak areas and ratio of hydrophobic to hydrophilic peptides, were all affected by the type of inoculum used as well as the strain under study. The results suggest that the accelerated-ripening model system developed, containing cell lysates, may be suitable as a good and rapid indicator of the contribution of the strains to proteolysis during cheese ripening.