家蝇抗菌肽MDC对鼠伤寒沙门氏菌的抑菌稳定性研究

以鼠伤寒沙门氏菌(Salmonella typhimurium)作为研究对象,探讨家蝇抗菌肽(Musca domestica cecropin,MDC)在不同条件下的抑菌稳定性。采用牛津杯法研究pH值、温度、光照、蛋白酶、金属离子、表面活性剂及有机试剂对MDC抑菌活性(抑菌圈大小)的影响。在酸性条件下MDC的抑菌活性稍有下降,保持在85%左右,但在碱性条件下的抑菌活性无明显改变,稳定性较好。经低温和高温处理后,抑菌活性无显著差异。MDC在黑暗、正常光及紫外线照射处理30 min,其抑菌活性仍保持稳定。MDC对胰蛋白酶敏感,抑菌活性完全消失,而对胃蛋白酶具有良好的稳定性。MDC对Ca²⁺、K⁺有良好的稳定性,但对Mg²⁺敏感,在Fe³⁺作用下其抑菌活性较对照组有所升高(P<0.05)。表面活性剂和有机试剂处理MDC后,其抑菌活性均明显降低。家蝇抗菌肽MDC能显著抑制鼠伤寒沙门氏菌生长繁殖,其活性几乎不受pH值,温度和光照变化、胃蛋白酶和某些金属离子的影响,且Fe³⁺能够协同...     

不同生境下的蒲公英及其不同器官的分段提取物抑菌作用比较研究

对不同生长环境下的蒲公英的根,叶,利用极性不同的有机溶剂分别进行萃取,比较各段萃取物的数量,分析其中有效物质的组分,并作抑菌试验,蒲公英叶较根萃取物多,阴生蒲公英的萃取的物较阴生蒲公英,其中氯仿所得萃取物抑菌效果最好,其主要含有生物碱,有机酸,甾醇等物质。     

蒲公英不同部位提取物抑菌作用研究及口服液研制

为检验蒲公英的抗菌活性并制备出口感最佳的蒲公英口服液,试验采用药敏法和最低抑菌浓度值(minimum inhibitory concentration,MIC)检验蒲公英提取物对大肠杆菌和金黄色葡萄球菌的抑菌效果,并以感官特性和黄酮含量为指标,并通过正交试验统计分析确定口服液的最佳工艺组方.结果表明:同浓度下全草水提物...     

抑制猪霍乱沙门氏菌的乳酸菌的分离、鉴定及潜在抑菌物质分析

猪霍乱沙门氏菌(Salmonella choleraesuis)是一种常见的食源性致病菌。为了预防和治疗该菌引起的疾病,饲养者在饲料中大量添加抗生素,致使猪肉存在严重的食品安全问题。从健康成年无量山乌骨鸡肠道内容物中筛选出具有抑菌作用的乳酸菌,测定其对猪霍乱沙门氏菌的抑菌效果,分析乳酸菌抑制猪霍乱沙门氏菌的有效成分,并对筛选的乳酸菌种进行了分子生物学鉴定。采用双层平板法对具有抑制猪霍乱沙门氏菌的乳酸菌进行筛选,采用牛津杯法对抑菌效果进行测定,并采用酶蛋白敏感性测定、热处理、有机酸处理等方法分析抑菌活性物质有效成分,采用16S rDNA分子标记对乳酸菌进行鉴定,并构建系统发育树。结果显示,从健康鸡肠道中筛选出18株乳酸菌,其中2株对猪霍乱沙门氏菌(Salmonella choleraesuis)、大肠埃希菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)、肠炎沙门氏菌(Salmonella enteritidis)、肠炎沙门氏菌亚种(Salmonella enteritidis subspecies)、志贺氏菌(Shigella)、无乳链球菌(Streptococcus agatactiae)具有良好的抑菌效果;不同蛋白酶、pH处理对乳酸菌无细胞培养液抑菌效果均有不同程度的影响,但是经80℃处理的乳酸菌无细胞培养液,其抑菌效果未明显改变。经鉴定,2株乳酸菌分别为植物乳杆菌(Lactobacillus plantarum)和短乳杆菌(Lactobacillus brevis)。从健康成年无量山乌骨鸡肠道内容物中分离得到的植物乳杆菌菌株L2和短乳杆菌菌株L4对猪霍乱沙门氏菌等致病菌具有明显地抑制作用,推测其抑菌有效成分可能为小肽类及有机酸,这对减少抗生素使用,提高猪肉食品的品质与安全性具有一定价值。     

短双歧杆菌对鼠伤寒沙门氏菌的抑制

【背景】鼠伤寒沙门氏菌是主要的肠道病原菌之一,利用益生菌治疗肠道病原菌感染已成为一种新型、绿色的微生态疗法。【目的】研究筛选出的短双歧杆菌无细胞发酵上清液(Cell-free supernatant,CFS)对鼠伤寒沙门氏菌的体外抑制作用及机制。【方法】采用微量稀释法测定短双歧杆菌YH68 CFS对鼠伤寒沙门氏菌的最小抑菌浓度(Minimum inhibitory concentration,MIC)和亚抑制浓度(Sub-inhibitory concentrations,SIC),并从鼠伤寒沙门氏菌的细胞形态、细胞膜通透性、膜完整性以及毒力基因表达的变化探讨YH68 CFS对鼠伤寒沙门氏菌的抑菌机理,同时检测YH68 CFS对鼠伤寒沙门氏菌粘附和侵袭肠上皮细胞HT29的影响。【结果】YH68 CFS (3×109 CFU/mL)对鼠伤寒沙门氏菌具有较好的抑制效果,抑菌圈直径为22.27±0.44 mm,最小抑菌浓度为250μL/mL,对鼠伤寒沙门氏菌的抑制机制是通过增加其细胞膜通透性破坏其完整性,形成难以修复的孔洞,最终达到抑菌的目的;亚抑制浓度为62.5μL/mL时YH68 CFS并不能影响鼠伤寒沙门氏菌的生长,但仍然能通过下调毒力基因表达的方式抑制其对肠上皮细胞的粘附和入侵。【结论】短双歧杆菌YH68对鼠伤寒沙门氏菌具有良好的抑菌作用,可作为治疗沙门氏菌感染的潜在益生菌。     

茶多酚对稻瘟病菌的抑制作用及抑菌机理

用不同浓度茶多酚对稻瘟病菌进行抑菌和抑菌机理研究。结果表明,不同浓度的茶多酚对稻瘟病菌菌丝生长和分生孢子萌发具有很强的抑制作用。随着茶多酚浓度的增加,其抑制作用增强,其中5.00 mg/mL和10.00 mg/mL抑制效果最好,其抑制率高达100%,且分生孢子畸形,细胞破裂,原生质外溢。其作用机理主要是破坏菌体的细胞膜结构,抑制CAT、POD酶活,使其丧失细胞膜的屏障和酶系的保护功能,最终导致菌体生长受到抑制或死亡。     

茶多酚对大肠杆菌抑菌机理的研究

茶多酚(tea polyphenols,TP)是一种具有广谱抗菌作用的活性质,以大肠杆菌(Escherichia coli,E.coli)为研究对象,采用牛津杯实验研究茶多酚的最小抑菌浓度,并采用结晶紫实验、电导率测定、离子泄漏测定来分析TP对E.coli细胞膜通透性的影响。通过琼脂糖凝胶电泳分析TP对E.coli DNA的影响。结果显示,TP对E.coli的最小抑菌浓度为40μg/m L;结晶紫实验中,随着浓度的增大,OD590也随之增大;随着处理时间的增长,电导率随之增大,离子泄漏随之增多,说明TP可以作用于E.coli的细胞膜,能够改变其通透性。通过琼脂糖凝胶电泳分析可知TP处理后和空白对照组相比较,DNA条带出现变暗、拖尾现象,说明TP能够作用于E.coli的遗传物质DNA。     

豆豉中高效抑菌活性乳酸菌的筛选及其抑菌研究

目的从云南传统发酵豆豉中分离乳酸菌,并从中筛选具有高抑菌活性的菌株,进一步研究其抑菌谱,并初步分析其抑菌机制,为新型生物防腐剂的开发提供理论依据。方法应用传统纯培养技术及spot-on-lawn法从云南传统发酵豆豉中筛选得到具有抑菌作用的乳酸菌40株,并从中筛选得到1株具有高效抑菌活性的植物乳杆菌YM-4-3菌株。通过HPLC和紫外分光对其主要抑菌物质进行分析与抑菌试验。结果 YM-4-3菌株是1株具有新型生物防腐剂应用潜力的乳酸菌菌株。结论初步推测YM-4-3菌株抑菌机制为有机酸主导,细菌素和其他化学抑菌物质协同作用。     

家蝇抗菌肽的抑菌动力学研究及其机理初探

利用鼠伤寒沙门氏菌针刺诱导家蝇幼虫表达抗菌肽,对抗菌肽的抑菌动力学进行研究,并通过抗菌肽样品对不同细菌动力学特性的研究出发对抗菌肽抑菌作用机制进行探讨。研究发现抗菌肽样品的活性与作用时间有关,24h内出现一到两个活性峰,同一抗菌肽样品对不同细菌的抑菌动力学有差异,抗菌肽的抑菌动力学机制应该与它的的抑菌作用机制有关。通过电镜观测、细胞磷代谢、紫外吸收物测定以及抗菌肽与细菌DNA相互作用结果可知,微生物诱导家蝇表达的抗菌肽样品不仅能够造成细菌细胞的快速坍塌破裂而且能够破坏细胞核心,与DNA结合作用。抗菌肽抑菌动力学的解释：微生物诱导产物中含有两类抗菌肽,一类抗菌肽能造成细胞膜的快速坍塌破裂形成第一个活性峰;另一类抗菌肽可进入细胞,破坏细胞核心,造成紫外吸收物大量外泄形成第二个活性峰。     

蓝莓抑菌活性研究进展

越橘属的蓝莓含有酚类物质、有机酸类、花青素类和糖类等生物活性成分。较早研究表明,蓝莓具有抗感染、抗氧化、抗肿瘤等功能,而近年的研究表明蓝莓具有抑菌作用。本文介绍了蓝莓的抑菌活性,分析了蓝莓中的抑菌活性成分,同时概括了蓝莓提取物对各种菌体的抑菌机理。     

Effects of phytic acid on the myoglobin-t-butylhydroperoxide-catalysed oxidation of uric acid and peroxidation of erythrocyte membrane lipids

Phytic acid stimulated the myoglobin-t-butylhydroperoxide (TBHP)-catalysed oxidation of uric acid, but inhibited the peroxidation of erythrocyte membrane lipids induced by the same system. Butylated hydroxy-toluene, a free radical chain reaction-terminating antioxidant, also suppressed the myoglobin-TBHP-induced lipid peroxidation. Moreover, phytic acid inhibited the hydroxyl radical-induced degradation of deoxyribose, but the extent of inhibition in this system was reduced by increasing the ferric ion concentration, suggesting that these effects of phytic acid on the myoglobin-TBHP-mediated oxidation are more likely attributable to its metal chelating properties rather than to a free radical scavenging action. The effectiveness of phytic acid, a naturally occurring antioxidant, in the inhibition of both iron- (as previously shown) and myoglobin-dependent lipid peroxidation suggests its possible therapeutic application as a non-toxic antioxidant for ameliorating the extent of oxy-radical-mediated myocardial ischemia/reperfusion damage.Abbreviations ASC Ascorbic acid - BHT Butylated Hydroxytoluene - DMSO Dimethyl Sulfoxide - TBHP t-Butylhydroperoxide - TBA Thiobarbituric Acid - TBARS Thiobarbituric Acid-reactive Substances     

低无机离子稻糠植酸提取方法的研究

采用醋酸解离并结合加热使植酸从种皮的复合盐中释放、石灰乳沉淀、乳酸酸化植酸钙、碱解再用离子交换脱无机离子,结合流程采用活性炭脱色的方法制备植酸.与同类方法相比,所得植酸纯度较高(68%～72%),总回收率达到70%以上;重复性及产品可溶性好,无非目的成分沉淀;色泽透明清亮,对优化粮油深加工的流程,提高产品质量有积极意义.     

壳聚糖抑菌机制的初步研究

壳聚糖在医学、食品、环保、日化用品等领域有着广泛而重要的应用.近年来,壳聚糖由于对不同的菌类都具有良好的抑菌效果而被研究者们密切关注.然而,有关壳聚糖抑菌机制的研究却并不多,其抑菌机制也没有被完全阐明.在本研究中,我们发现很多金属离子可以对壳聚糖的抑菌效果产生影响,高浓度金属离子(0.5%)可以使壳聚糖完全丧失抑菌活性.还发现金黄色葡萄球菌和白色念珠菌在壳聚糖的作用下会发生钾离子和ATP的渗漏,而且五万分子量的壳聚糖引起钾离子和ATP的渗漏大约比五千分子量壳聚糖多2到4倍.不同分子量的壳聚糖对金黄色葡萄球菌和白色念珠菌都具有较好的抑菌效果,但是引起钾离子和ATP的渗漏量却存在很大差异,这说明小分子量壳聚糖很可能存在与大分子量壳聚糖不同的抑菌机制.     

木犀草素对金黄色葡萄球菌的抑菌活性及其机制

【目的】研究木犀草素对金黄色葡萄球菌的抑制活性及其机制。【方法】利用2,3,5-氯化三苯基四氮唑(TTC)染色,细胞膜渗透性测定,SDS-PAGE蛋白谱变化,4′,6-二脒基-2-苯基吲哚(DAPI)荧光染色法等对木犀草素的抑菌活性及其机制进行研究。【结果】木犀草素能影响金黄色葡萄球菌细胞膜的通透性,木犀草素作用16h,菌体可溶性蛋白总量减少64.54%,DNA含量减少48.44%,RNA含量减少39.35%,木犀草素的浓度为1.6mg/mL时,拓扑异构酶I和II的活性可完全被抑制。【结论】木犀草素有明显的抑菌活性,其抑菌机制主要是通过抑制DNA拓扑异构酶的活性,进而影响菌体核酸及蛋白质的合成来实现的。     

Harvesting of Arthrospira platensis by flocculation with phytic acid from rice bran

ABSTRACT

The recovery of algal biomass is one of the critical steps involved in the commercial production of beneficial metabolites from Arthrospira platensis. Efficient and safe harvesting methods that do not sacrifice quality of final product are important for commercial application. Phytic acid (PA) is a natural non-toxic phytochemical widely distributed in plant tissues. Effect of PA from rice bran on the growth, trichome morphology such as spiral number and algal filament length, and harvesting efficiency of A. platensis were investigated. Cells aggregated into large cell flocs after the addition of PA in the medium, and algal spiral number and filament length increased. UV-vis spectra indicated the interactions between PA and algal cells. Adding PA at stationary growth phase is a good strategy for harvesting, since no adverse effect to biomass growth and harvesting efficiency. Harvesting efficiency of 95.69% at 0.5% (v/v) PA was superior to other conventional harvesting methodologies.     

Synchronous fluorescence determination of phytic acid in FOODSTUFFS and urine based on replacement reaction

Introduction – Phytic acid is a ubiquitous and abundant natural component in many plant seeds, fruits and vegetables. Its biological and pharmaceutical functions are still controversial. The examination on the level of phytic acid in foodstuffs and urine can provide valuable information for its dietary intake and metabolism. Objective – To develop a sensitive and reliable synchronous fluorescence protocol for determination of phytic acid in selected foodstuffs and human urine. Methodology – Phytic acid efficiently catches Cu²⁺ ion in previously prepared Cu^II‐2,2′‐bipyridine complex in aqueous solution, releasing the fluorescent 2,2′‐bipyridine molecule and recovering synchronous fluorescence. The recovered fluorescence is proportional to the added phytic acid, by which the levels of phytic acid in the selected foodstuffs and human urine are quantified. Results – A calibration curve with a regression equation of I_f = 37.745 + 39.245c (R² > 0.9988) showed good linearity over the range 0.18–17.50 mg/L phytic acid. The relative standard deviation at 95% confidence degree was less than 2.04% (n = 5), indicating that the procedures are reproducible. The detection and quantification limit of phytic acid were estimated to be 0.12 and 0.18 mg/L, respectively. By the proposed method, phytic acid in the selected foodstuffs and urine was determined to be 3.25–16.76 and 0.43–1.21 mg/L with recoveries of 96.8%–105.6% and 95.1%–104.2%, respectively. The results are in good agreement with those obtained by the reported HPLC technique. Conclusion – The developed method is sensitive, reliable and economical, which permits its practical application in quantitative analyses of trace phytic acid in foodstuffs and urine. Copyright © 2010 John Wiley & Sons, Ltd.     

大黄酸抗耐甲氧西林金黄色葡萄球菌的抗菌机制

[背景]耐甲氧西林金黄色葡萄球菌(Methicillin-Resistant Staphylococcus aureus,MRSA)是医院及社区常见的机会性致病菌,具有多重耐药性、高发病率和高死亡率的特点.MRSA感染已成为全球医学界的普遍难题之一.[目的]研究大黄酸对MRSA的抗菌机制.[方法]以二倍稀释法测定大黄酸...     

Ferric ion-induced lipid peroxidation in erythrocyte membranes: effects of phytic acid and butylated hydroxytoluene

Ferric ion was found to stimulate the peroxidation of erythrocyte membrane lipids, causing a biphasic and concentration-dependent increase in the formation of thiobarbituric acid reactive substances. Ascorbic acid and reduced glutathione were able to enhance this lipid peroxidation, presumably by facilitating the reduction of ferric ion. Iron chelators, such as phytic acid, ethylenediaminetetraacetic acid and uric acid, and the chain-reaction-terminating antioxidant butylated hydroxytoluene suppressed the ferric ion-induced peroxidation by actions not likely related to hydroxyl radical scavenging. The effectiveness of phytic acid, a naturally occurring antioxidant, in the inhibition of iron-dependent lipid peroxidation suggests its possible therapeutic application as a non-toxic iron chelator for ameliorating the extent of oxy-radical-induced tissue damage.Abbreviations BHT Butylated Hydroxytoluene - EDTA Ethylenediaminetetraacetic Acid - GSH Reduced Glutathione - TBA 2-Thiobarbituric Acid - TBARS 2-Thiobarbituric Acid Reactive Substances     

Increased erythritol production in Torula sp. with inositol and phytic acid

Of the vitamins tested, inositol was the most effective for erythritol production. To increase erythritol production by Torula sp., inositol and a related compound, phytic acid (myoinositol hexaphosphate), were added to the culture media. Erythritol production in the presence of phytic acid was greater than that in the presence of inositol, due to the synergistic effects of phosphate and inositol. Supplementation with phosphate and inositol increased cell growth, erythritol production, and the activity of erythrose reductase in cells. Inositol was a more effective stimulator of cell growth and erythritol production than was phosphate.     

Evaluation of alternatives for human lysozyme purification from transgenic rice: impact of phytic acid and buffer

Producing economically competitive recombinant human lysozyme from transgenic rice demands an inexpensive purification process for nonpharmaceutical applications. Human lysozyme is a basic protein, and thus, cation exchange chromatography was the selected method for lysozyme purification. Similar to other protein production systems, the identification of critical impurities in the rice extract was important for the development of an efficient purification process. Previous adsorption data indicated that phytic acid was probably responsible for an unacceptably low cation exchange adsorption capacity. In this study, we confirm that reducing phytic acid concentration improves lysozyme binding capacity and investigate alternative process conditions that reduce phytic acid interference. Compared with the previous best process, the adsorption capacity of human lysozyme was increased from 8.6 to 19.7 mg/mL when rice extract was treated with phytase to degrade phytic acid. Using tris buffer to adjust pH 4.5 extract to pH 6 before adsorption reduced phytic acid interference by minimizing phytic acid-lysozyme interactions, eliminated the need for phytase treatment, and increased the binding capacity to 25 mg/mL. Another method of reducing phytic acid concentration was to extract human lysozyme from rice flour at pH 10 with 50 mM NaCl in 50 mM sodium carbonate buffer. A similar binding capacity (25.5 mg/mL) was achieved from pH 10 extract that was clarified by acidic precipitation and adjusted to pH 6 for adsorption. Lysozyme purities ranged from 95 to 98% for all three processing methods. The tris-mediated purification was the most efficient of the alternatives considered.