风干羊肉中乳酸菌的体内外抗氧化特性

【背景】机体的衰老和一些疾病多与氧化作用有关,随着对抗氧化制剂研究的深入,人工合成抗氧化剂的安全性受到质疑,因此寻找天然抗氧化制剂的研究已成为热点。【目的】从风干羊肉中筛选抗氧化活性较高的乳酸菌,分析肉源乳酸菌体内外抗氧化能力。【方法】以24株肉源乳酸菌为研究对象,以自由基清除能力、亚铁离子螯合能力、还原能力及抗脂质过氧化能力为体外抗氧化能力分析指标,筛选出体外抗氧化活性较强的乳酸菌,运用16S rRNA基因序列同源性分析进行菌种鉴定,通过小鼠试验研究其体内抗氧化能力。【结果】试验所测24株乳酸菌均具有一定的体外抗氧化能力,其中菌株TR13为24株乳酸菌中体外抗氧化能力较强的菌株,其超氧阴离子自由基(·O2?)清除率为54.29%,羟自由基(·OH)清除率为90.84%,1,1-二苯基-2-三硝基苯肼自由基(1,1-diphenyl-2-picrylhydrazyl,DPPH)清除率为99.38%,亚铁离子螯合率为55.85%,还原能力达1.345,抗脂质过氧化率为39.99%。通过16S rRNA基因鉴定,菌株TR13为一株瑞士乳杆菌。通过给D-半乳糖诱导的衰老型小鼠饲喂TR13菌液,验证了TR13对小鼠的肝脏、肾脏及血液组织中谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性、超氧化物歧化酶(superoxide dismutase,SOD)活性、过氧化氢酶(catalase,CAT)活性均有显著的提升作用(p<0.05),且TR13组的GSH-Px活性显著高于维生素C (Vc)组(p<0.05)。【结论】菌株TR13对机体的氧化具有一定的防御作用,研究结果可为乳酸菌抗氧化制剂的研究提供参考依据。     

两株乳酸杆菌SY13和LJJ对活性氧的耐受性

【目的】从传统发酵乳制品中筛选具有抗氧化能力的乳酸菌菌株并对其抗氧化特性进行评价。【方法】分别利用乳酸杆菌SY13和LJJ完整细胞和无细胞提取物对亚油酸过氧化的抑制效果,对DPPH自由基、羟自由基、超氧阴离子自由基清除能力,对过氧化氢的耐受性以及对亚铁离子的螯合能力和还原活性进行了研究。【结果】结果表明,SY13和LJJ对亚油酸过氧化的最大抑制率分别达到了62.95%和66.16%;两菌株的无细胞提取物清除超氧阴离子和羟自由基的的效果较好,LJJ完整细胞对超氧阴离子和羟自由基没有清除能力;SY13和LJJ对DPPH自由基的清除能力及对亚铁离子的螯合能力都是完整细胞优于无细胞提取物,还原活性分别相当于305 μmol/L和294 μmol/L的L-cysteine。【结论】以上指标测定的结果说明,这两株乳酸杆菌具有较好的抗氧化能力,具有潜在的应用价值。     

层孔菌属真菌抗氧化活性物质测定及抗氧化能力分析

本文以层孔菌属真菌液体发酵产物作为研究材料,测定抗氧化活性物质含量,并利用多种抗氧化测定方法,分析不同菌株的体外抗氧化能力,评价活性物质含量间、抗氧化方法间及抗氧化方法与含量间的相关性。研究结果表明,供试菌株活性物质含量间的差异较大,其中菌株H2粗多糖和总酚含量较高,菌株S黄酮含量较高,且活性物质含量间相关性不大;针对供试菌株抗氧化能力测定结果表明,菌株H2具有较高的还原力、铁离子螯合能力,菌株H2和H9的羟自由基清除作用最强,菌株H2和L的DPPH自由基清除作用最强,菌株H2、H5、H8、H9具有较高的超氧自由基清除作用,且不同方法间存在较大差异,羟自由基与DPPH自由基、铁离子螯合能力及还原力,DPPH自由基与铁离子螯合能力和还原力,以及铁离子螯合能力与还原力这几种方法的相关性较高(P0.01);总酚含量与羟自由基、DPPH自由基、铁离子螯合能力及还原力方法具有较高的相关性(P0.01);总黄酮含量与DPPH自由基、铁离子螯合能力及还原力方法间相关系数分别为0.810(P0.01)、0.725、0.738(P0.05);粗多糖含量与各测定方法间的相关系数较低,以上结果说明多酚类物质和黄酮类物质是抗氧化作用的重要因子,对抗氧化能力具有重要贡献。     

药用真菌灵芝液体培养过程中的抗氧化活性研究

灵芝Ganoderma lingzhi是一种重要的药用真菌,已被《中国药典》正式收录。本研究主要以菌丝体干重、多糖含量、多酚含量、黄酮含量、抗坏血酸（ascorbic acid,AA）含量、总抗氧化能力（total antioxidant capacity,T-AOC）、超氧化物歧化酶（superoxide dismutase,SOD）活性、羟自由基清除能力、超氧阴离子清除能力、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除能力、2,2′-连氮-双(3-乙基苯并噻唑-6-磺酸)[2,2′-azino-bis(3- ethylbenzthiazoline-6-sulphonic acid),ABTS]自由基清除能力、铁离子还原能力（ferric reducing antioxidant power,FRAP）和亚铁离子螯合能力为测定指标,对灵芝液体培养过程中的抗氧化活性进行了评价。结果显示,该菌具有较高的抗氧化活性,体现在液体培养过程中生长代谢旺盛,可分泌大量多糖、多酚、黄酮、AA等物质和SOD等酶类,对羟自由基、超氧阴离子、DPPH自由基及ABTS自由基等的清除效果显著,且具有较强的铁离子还原能力和亚铁离子螯合作用,这也说明该菌的抗氧化活性与其自身的生长状况、次级代谢产物分泌及还原能力等密切相关。此外,一定的环境胁迫压力也可以激发该菌启动自身的抗氧化系统以保护机体免受氧化损伤。     

乳酸菌抑制葡聚糖硫酸钠诱导的结肠黏膜氧化应激损伤的研究

目的研究乳酸菌抗氧化活性与缓解葡聚糖硫酸钠(DSS)诱导结肠炎的关系。方法评价5株乳酸菌体外抑制大鼠肝细胞脂质过氧化活性,用DSS诱导小鼠结肠炎模型研究抗氧化性不同的菌株体内抗氧化活性;小鼠分成7组,分别是Control组、DSS组、L.plantarum Fn008+DSS组、L.casei Fn012+DSS组、L.acidophilus Fn022+DSS组、L.sakei Fn034+DSS组和L.acidophilus Fn037+DSS组。结果体外实验表明,Fn022和Fn034的丙二醛(MDA)抑制率高于LGG,但差异无统计学意义,Fn008和Fn037的MDA抑制率显著低于LGG,Fn012的MDA抑制率最低。体内实验表明,DSS可导致小鼠氧化应激,升高结肠内容物的自由基水平,降低结肠抗氧化能力;Fn008和Fn037预处理能显著降低DSS诱导结肠内容物的自由基水平,Fn008能显著降低结肠组织的髓过氧化物酶(MPO)水平,提高结肠抗氧化能力,Fn022和Fn034的干预能力居中,Fn012的干预能力最弱。相关性分析表明乳酸菌体外MDA清除率与体内MPO有负相关趋势(r=-0.862,P=0.06)。结论乳酸菌抗氧化活性与其在肠道抑制炎性细胞激活的作用有关,本实验中Fn008和Fn037具有显著的体内抗氧化作用。     

连翘乙酸乙酯提取物体外抗氧化活性研究

目的:研究连翘乙酸乙酯提取物的体外抗氧化活性,为其充分开发利用提供理论依据。方法:以乙酸乙酯为溶剂制备连翘乙酸乙酯提取液,通过分光光度法测定其对DPPH自由基的清除能力、用普鲁士蓝法测定其还原Fe~(3+)的能力,并以丁基羟基茴香醚(BHA)为对照品测定连翘乙酸乙酯提取物对金属亚铁离子(Fe~(2+))的螯合能力。结果:连翘乙酸乙酯提取物具有较强的DPPH自由基的清除能力和还原Fe~(3+)能力,对亚铁离子(Fe~(2+))的螯合能力强。结论:连翘乙酸乙酯提取物的抗氧化能力强,且随样品液的浓度升高而增强,呈现良好的线性关系。     

药用真菌桑黄液体培养过程中的抗氧化活性研究

桑黄Sanghuangporus sanghuang是一种珍稀的药用真菌,具有较大的应用潜力。本研究主要以菌丝体生物量、多酚含量、黄酮含量、抗坏血酸(Ascorbic acid,AA)含量、总抗氧化能力(Total antioxidant capacity,T-AOC)、抑制羟自由基能力(Restraining ability to hydroxyl free radicals,RAHFR)、超氧阴离子清除能力、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除能力、2,2’-连氮-双(3-乙基苯并噻唑-6-磺酸)[2,2’-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid),ABTS]自由基清除能力、铁离子还原能力(Ferric reducing antioxidant power,FRAP)和亚铁离子螯合能力为测定指标,对桑黄液体培养过程中的抗氧化活性进行了评价。结果显示,菌丝体生物量在2–10d内增长迅速,第10天达到最大值,在此过程中AA含量和T-AOC也出现了峰值,且多酚含量、黄酮含量和FRAP均呈上升趋势,说明菌株的抗氧化活性与其自身的生长状况、次级代谢产物分泌及还原能力等密切相关。此外,该菌株对羟自由基、超氧阴离子、DPPH自由基和ABTS自由基的清除能力也较强,于第12天、第10天、第2天和第14天分别达到73.06U/m L、46.78%、86.47%和94.16%,显示了较强的抗氧化活性。较高的亚铁离子螯合能力也说明桑黄在氧化胁迫下可启动自身的抗氧化系统以阻断自由基链反应。研究结果为更好地研究、开发和利用药用真菌桑黄提供了基础资料。     

平菇菌粗多糖的抗氧化活性研究

采用深层发酵技术生产平菇粗多糖,时其清除DPPH自由基、羟自由基的能力、铁离子螯合能力以及还原力进行了比较分析。结果表明：菌丝体粗多糖和发酵液粗多糖均具有较强的抗氧化能力,但2种多糖的抗氧化能力存在差异;茵丝体粗多糖清除DPPH自由基的能力较强,其EC。。值为2．20mg／mL;发酵液粗多糖清除羟自由基的能力、铁离子螯合能力以及还原力较强,其EC50值分别为0．72mg／mL、3．32mg／mL和7．91mg／mL。在一定的浓度范围内,多糖的浓度增加,其抗氧化能力也随之增强,呈量效依赖关系。     

消化乳杆菌泡菜分离株W369的鉴定及其益生功能北大核心CSCD

【目的】鉴定从中国传统发酵泡菜分离的1株乳酸菌,并分析其潜在的益生功能。【方法】通过形态观察、生理生化、16S rRNA基因序列分析等方法对分离得到的乳酸菌进行鉴定;评价该菌株的NaCl耐受能力、抗生素最小抑制浓度,以及益生功能。【结果】该菌株鉴定为消化乳杆菌(Lactobacillus alimentarius)W369,在10%的NaCl浓度下仍可以存活;对常用抗生素具有不同程度的敏感性。益生功能分析表明,培养72 h后W369对亚硝酸钠的降解率为92.92%;其体外胆固醇脱除率达到31.80%;发酵液上清具有清除DPPH自由基(DPPH·)、超氧阴离子自由基(·O_2~–)和螯合亚铁离子(Fe^(2+))的作用,分别达到(88.02±1.48)%、(43.75±3.10)%和(29.99±2.34)%;完整细胞具有很强的还原能力[(244.52±0.92)μmol/m L半胱氨酸当量]、羟自由基(·OH)清除能力(77.17±0.58)%和抗脂质过氧化的能力(29.67±0.77)%;发酵上清液和无细胞提取物均具有谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(Mn-SOD)活性。【结论】消化乳杆菌W369分离株具有亚硝酸盐降解能力、胆固醇清除能力和抗氧化活性等诸多益生功能,具有潜在的应用价值。     

樟芝发酵液和菌丝体提取物抗氧化性能研究

采用甲醇、乙醇、异丙醇对樟芝发酵液和菌丝体进行提取,对其不同提取物进行还原力,DPPH、羟基自由基、超氧自由基、过氧化氢清除能力,亚铁离子螯合能力等抗氧化能力的测定。比较发酵液提取物和菌丝体提取物的抗氧化效果,比较不同提取剂提取物的抗氧化效果。结果显示:(1)发酵液提取物还原力最高的为异丙醇提取物,亚铁离子螯合能力最强的为乙醇提取物;DPPH清除能力乙醇和异丙醇提取物最高都达到97.88%;羟基自由基清除能力最高的为乙醇提取物,达到84.22%;3种提取物的超氧自由基清除能力都较高,具有最高清除率的异丙醇提取物达到94.89%;过氧化氢清除能力最高的为乙醇提取物的97.53%。(2)菌丝体提取物还原力最高和亚铁离子螯合能力最强的均为乙醇提取物;DPPH清除能力最高的为乙醇提取物的89.95%;3种提取物的羟基自由基清除能力普遍偏低,最高的仅为异丙醇提取物的37.28%;超氧自由基清除能力则普遍较高,最高为甲醇提取物的90.05%;过氧化氢清除能力最高的为乙醇和异丙醇提取物,最高清除率均为95.06%。抗氧化性能的研究比较,为进一步开发利用樟芝提供了依据。     

Inhibition of Fe-induced colon oxidative stress by lactobacilli in mice

Iron (Fe) can promote hydrogen peroxide (H₂O₂) and hydroxyl radical generation in the colonic surface and promote growth of Fe-dependent bacteria. Some Lactobacillus strains are resistant to oxygen free-radicals, allowing them to survive in a Fe-modulated mucosal environment and influence colon microbial ecology and redox state. Here, we investigated the capacity of lactobacilli with different antioxidant abilities to modify the bacterial profile and prevent oxidative stress in the colon of Fe-overloaded mice. Survival time of Lactobacillus rhamnosus LGG (LGG) in the presence of H₂O₂ and hydroxyl radical was significantly longer compared with the mid- and non-antioxidative strains, Lactobacillus paracasei Fn032 and Lactobacillus plantarum Fn001, respectively. Different Lactobacillus strains are specific in free-radical scavenging activities of their cell-free extracts, which increased to varying extent depending on strains when bacteria were exposed to simulated gastric and pancreatic juice. Fe-overloaded mice showed increased colonic luminal ferrous Fe content, Enterococcus and Escherichia coli concentrations, mucosal malondialdehyde and free-radicals, and decreased mucosal total antioxidative capacity and oxidative enzymatic activity. Translocation of endotoxin to the liver was also significantly increased (P < 0.05). Lactobacilli inhibited ferrous Fe accumulation, especially in LGG and Fn032. LGG significantly inhibited the increase of colonic mucosal free-radicals and malondialdehyde content (P < 0.05). Fn032 only inhibited malondialdehyde (P < 0.05). LGG and Fn032 significantly inhibited increases in colonic Enterococcus (P < 0.05). Fn001 showed no significant antioxidative ability in vivo. The difference of these effects in vivo were well agreed with scavenging activities against reactive oxygen species (ROS) of simulated gastrointestinals fluid pretreated cells in vitro. In conclusion, ROS scavenging activities was essential for Lactobacillus to prevent oxidative stress in vivo and inhibition of ROS-producing bacterial growth and mucosal barrier injury.     

Lactobacilli prevent hydroxy radical production and inhibit Escherichia coli and Enterococcus growth in system mimicking colon fermentation

Aims: To explore the effect of Lactobacillus on redox state of colon chyme. Methods and Results: Nine Lactobacillus strains were studied for the inhibition of lipid peroxide formation in Fe²⁺/ascorbate system and for their ability to chelate ‘free’ ferrous ion. The result shows both properties were strain specific and no relationship between them was found. Both properties of Lactobacillus paracasei Fn032, Lactobacillus rhamnosus GG (LGG) and Lactobacillus sp. Fn001 were successively decreasing. LGG and Fn032 significantly decreased hydroxyl radicals (P < 0·01) in colonic fermentation model, in which considerable hydroxyl radicals occurred spontaneously. Addition of ferrous ion induced the production of hydroxyl radicals, which could be significantly inhibited by LGG, Fn032 (P < 0·01) and Fn001 (P < 0·05). Ferrous ion significantly induced the growth of Enterococcus and Escherichia coli, which could be inhibited by all three Lactobacillus strains. Escherichia coli and Enterococcus show significantly positive correlation with hydroxyl radicals with R of 0·96 (P = 0·0002) and 0·91 (P = 0·0017), respectively. Conclusions: Antioxidative Lactobacillus could modulate redox state in colonic fermentation system, which is related to their free radical–scavenging ability or antibacterial effect. Significance and Impact of the Study: This study proves that Lactobacillus strain could influence the redox state of gut chyme. Evaluation of antioxidative ability might be a powerful method for screening probiotic Lactobacillus strains.     

南瓜多糖衍生物的制备及其抗氧化研究

对超声辅助法提取的南瓜粗多糖,进行化学改性得到了硫酸化南瓜多糖和羧甲基南瓜多糖。用邻苯三酚自氧化法检测对超氧阴离子自由基的清除作用,用邻二氮菲-金属铁离子-H2O2体系检测对羟自由基的清除作用。结果表明,南瓜多糖对超氧阴离子自由基无抑制作用,经化学改性后的羧甲基南瓜多糖和硫酸化南瓜多糖对超氧阴离子自由基有明显抑制作用,最大清除率达44％和41．67％。以上3种多糖均能有效清除·OH,随着浓度的增加清除作用加强,最大清除率可达46．49％,22．61％和42．35％0。结果显示,经化学改性后的南瓜多糖有较强的抗氧化作用。     

Quantitative evaluation of adhesion of lactobacilli isolated from human intestinal tissues to human colonic mucin using surface plasmon resonance (BIACORE assay)

AIMS: To isolate lactobacilli from the mucus layer of the human intestine and evaluate their adhesion abilities using a BIACORE assay. METHODS AND RESULTS: Thirty strains of lactobacilli were isolated from the mucus layer of normal human intestinal tissues using conventional plate culture. The strains were identified using homology comparisons of the 16S rDNA sequence to databases as Lactobacillus salivarius (26%), Lactobacillus fermentum (13%), Lactobacillus gasseri (10%), Lactobacillus paracasei (7%), Lactobacillus casei (3%), Lactobacillus mucosae (3%) and Lactobacillus plantarum (3%). Lactobacillus plantarum LA 318 shows the highest adhesion to human colonic mucin (HCM) using the BIACORE assay at 115.30 +/- 12.37 resonance unit (RU). The adhesion of cell wall surface proteins from strain LA 318 was significantly higher to HCM than to bovine serum albumin (BSA; P < 0.05). CONCLUSIONS: We isolated 30 strains of lactobacilli. Lactobacillus salivarius was the predominant species of lactobacilli isolated in this study. The adhesion of strain LA 318 isolated from human transverse colon to its mucin was shown. The adhesion could be mediated by lectin-like components on the bacterial cell surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study where lactobacilli were isolated from human intestinal tissues and shown to adhere to HCM.     

Characterization of intestinal lactobacilli as putative probiotic candidates

AIMS: To use antioxidative activity and antagonistic properties of lactobacilli against selected pathogens and members of the normal microflora as a basis for screening probiotic candidates. METHODS AND RESULTS: Antagonistic activity of lactobacilli against target bacteria in both microaerobic and anaerobic environments was tested. Production of antagonistic metabolites (ethanol, hydrogen peroxide (H2O2), acetic, lactic and succinic acid) by lactobacilli as well as their total antioxidative activity were assessed. In general, the lactobacilli tested were most effective against Gram-negative bacteria and their antagonistic activity was strain-specific. However, obligately heterofermentative lactobacilli had the strongest activity when tested in a microaerobic environment. Additionally, facultatively heterofermentative lactobacilli were equally effective in either milieu and produced significant levels of acetic and lactic acid. Moreover, obligately homofermentative lactobacilli had high H2O2 production and total antioxidative activity but weak antagonistic activity. CONCLUSIONS: Antioxidative and antagonistic activity of intestinal lactobacilli is strain-specific but typically can be related to their fermentation type which may be used for rapidly screening large numbers of lactobacilli for probiotic candidates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents the first report on the utilization of group characteristics to screen lactobacilli intended for specific probiotic use. Such uses include the targeting of particular gut niches and pathogens as well as allowing for long-term benefits to the host.     

尾叶远志抗氧化活性研究

采自贵州兴义的尾叶远志(Polygala caudata Rehd.et Wils.)根,经乙醇提取,再用不同溶剂萃取和柱层析分离,得到了12个不同的组分.用比色法研究了各组分还原三价铁离子及清除脂性自由基DPPH能力,采用化学发光法观察各组分清除羟自由基和超氧阴离子的活性.研究发现,尾叶远志各组分均有一定的清除自由基作用,并且其活性成分可能主要集中在PC-BuOH组分.     

Development of antioxidant activity in milk whey during fermentation with lactic acid bacteria

AIMS: To investigate the production of antioxidant activity during fermentation with commonly used dairy starter cultures. Moreover, to study the development of antioxidant activity during fermentation, and the connection to proteolysis and bacterial growth. METHODS AND RESULTS: Antioxidant activity was measured by analysing the radical scavenging activity using a spectrophotometric decolorization assay and lipid peroxidation inhibition was assayed using liposomal model system with a fluorescence method. Milk was fermented with 25 lactic acid bacterial (LAB) strains, and from these six strains, exhibiting the highest radical scavenging activity was selected for further investigation. Leuconostoc mesenteroides ssp. cremoris strains, Lactobacillus jensenii (ATCC 25258) and Lactobacillus acidophilus (ATCC 4356) showed the highest activity with both the methods used. However, the radical scavenging activity was stronger than lipid peroxidation inhibition activity. The development of radical scavenging activity was connected to proteolysis with four strains. Molecular distribution profiles showed that fermentates with high scavenging activity also possessed a higher proportion of peptides in the molecular mass range of 4-20 kDa, while others had mostly large polypeptides and compounds below 4 kDa. In addition, the amount of hydrophobic amino acids was higher in these fermentates. CONCLUSIONS: The development of antioxidant activity was strain-specific characteristic. The development of radical scavengers was more connected to the simultaneous development of proteolysis whereas, lipid peroxidation inhibitory activity was related to bacterial growth. However, high radical scavenging activity was not directly connected to the high degree of proteolysis. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this seems to be the first report, which screens possible antioxidant activity among most common dairy LAB strains. Use of such strains improve nutritional value of fermented dairy products.     

Anti-oxidation of agar oligosaccharides produced by agarase from a marine bacterium

In order to prepare the active agar oligosaccharide, agarase extracted from a strain of unidentified marine bacterium from the South China Sea coast was selected for the agar depolymerization. The optimum decomposing conditions were determined to be pH 7.0, 35 °C and halophilic properties 2%. Three main degraded products, AOS-1, AOS-2 and AOS-3, were separated by ethanol fractionation and anion exchange chromatography. The molecular mass was analyzed by MALDI-TOF-MS. The agar oligosaccharides exhibited antioxidative activities in scavenging hydroxyl free radical, scavenging superoxide anion radical and inhibiting lipid peroxidation. The fragment with the sulfate group showed stronger antioxidative activities than that without the sulfate group. Higher antioxidative activities were found when the molecular mass was increased. The results indicated that the antioxidative activities were closely related to the molecular mass of the agar oligosaccharides and the substitute groups binding the carbohydrate ring.