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Study of Antibacterial Substances Found in Natto

纳豆中抗菌物质的研究

基本信息

批准号：
15500558
负责人：
SUMI Hiroyuki
金额：
$ 2.3万
依托单位：
Kurashiki University of Science and The Arts
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

An analysis of dipicolinic acid (DPA,2,6-pyridinedicarboxylic acid) contained in Bacillus subtilis natto cells using the colorimetric method of Janssen and others showed that the largest DPA content is approximately 2.4-3.6% of the total dry weight.A comparison of the antibacterial activity of DPA and its derivatives-quinolic acid (2,3-pyridine dicarboxylic acid), 2,4-pyridine dicarboxylic acid, isoinchlorendic acid (2,5-pyridine dicarboxylic acid), and 3,5-pyridine dicarboxylic acid-revealed that this antibacterial activity is unique to DPA. Moreover, an investigation into the effects of various metal ions on the antibacterial activity showed that the activity is strongly inhibited through the addition of Ca^<++>,Fe^<++>,Co^<++>, and Zn^<++>, all divalent ions.It was also shown that the addition of DPA increases the amount of nattokinase produced by Bacillus subtilis natto. In four types of Bacillus subtilis natto, the Miyagino, Meguro, Takahashi and Naruse types, DPA with a 2 mM conc … More entration increased the amount of nattokinase produced. In the case of the Miyagino type, for example, the fibrinolytic activity for fibrin plates was double the level seen for the sample with no DPA added, at 256.1 mm^2/30 μl at 37℃ after four hours (1,500 FU/ml for the nattokinase activity). A comparison test, with 10 types of nicotinic acid-related substances added, showed that dipicolinic acid had the strongest effect.DPA with a final concentration of 5×10^<-3> mol/l was found to substantially inhibit the platelet aggregation reaction caused by human adenosine triphosphate(ATP), as well as the reaction of thrombin converting fibrinogen into fibrin. It became clear that at 5×10^<-3> mol/l, there was a complete inhibition of the blood coagulation reaction as observed in thromboelastography, while such strong effects were not observed in the case of DPA derivatives, even at 10 mM. There were no changes in human prothrombin time, the thromboplastin time for the active areas, or euglobulin lysis time.Although platelet aggregation in human blood is observed with 30 μM of adenosine diphosphate(ADP), it became clear that dose-dependent inhibition of this aggregation occurs when there is 5×10^<-3> M of DPA. The calculated CI_<50> was at the fairly low concentration of 1.8×10^<-3> M, and far stronger (4.0×10^<-2> M) than in the case of adding aspirin for clinical use. Less

用Janssen等人的比色法对纳豆芽孢杆菌细胞中含有的二吡啶甲酸(DPA)进行了分析，结果表明，DPA的含量最高，约占总干重的2.4-3.6%。比较了DPA及其衍生物--喹酚酸(2，3-吡啶二酸)、2，4-吡啶二酸、异氯酸(2，5-吡啶二酸)和3，5-吡啶二酸--的抗菌活性，发现DPA是DPA所特有的。此外，研究了不同金属离子对纳豆芽孢杆菌抗菌活性的影响，结果表明，二价金属离子Ca^&lt；++&gt；、Fe^&lt；++&gt；、Co^&lt；++&gt；和Zn^&lt；++&gt；均对纳豆菌的抗菌活性有较强的抑制作用，DPA的加入使纳豆菌产生的纳豆激酶量增加。在四种类型的纳豆枯草芽孢杆菌中，宫野型、美黑型、高桥型和纳鲁斯型的DPA带有2 mM的conc…更多的进入增加了纳豆激酶量的产生。例如，在宫野型的情况下，纤维蛋白平板的纤溶活性是不添加DPA的样品的两倍，在256.1 mM^2/30μL，4小时后37℃(纳豆酶活性为1,500 fU/ml)。添加10种烟酸相关物质的对比试验表明，二吡啶甲酸的作用最强。最终浓度为5×10~(-3)mol/L的DPA可显著抑制人三磷酸腺苷引起的血小板聚集反应，以及凝血酶将纤维蛋白原转化为纤维蛋白的反应。结果表明，在5×10~(-3)~(-3)mol/L时，血栓弹性图法所观察到的凝血反应被完全抑制，而在DPA衍生物的情况下，即使在10 mM时也没有观察到如此强烈的作用。人凝血酶原时间、活性部位的凝血活酶时间或优球蛋白溶解时间没有变化。虽然30μM的二磷酸腺苷可以观察到人血中的血小板聚集，但很明显，当DPA的浓度为5×10~(-3)M时，这种聚集的抑制作用是剂量依赖性的。计算出的CI_50&gt；处于相当低的浓度1.8×10；-3&gt；M，比临床使用添加阿司匹林的情况强得多(4.0×10；-2&gt；M)。较少