STRUCTURE AND CHARACTERISTICS OF WATER-SOLUBLE ANTIMICROBIAL SUBSTANCES FORMED BY REACTION OF ISOTHIOCYANATES WITH AMINO ACIDS

异硫氰酸酯与氨基酸反应形成的水溶性抗菌物质的结构和特性

• 批准号: 04660131
• 负责人: UDA Yasushi
• 金额: $ 0.32万
• 依托单位: UTSUNOMIYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C).
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04660131/
• 关键词: Antimicrobial activity; Mustart Oil; Amino acid; 芥子油類; 抗菌性物質

The project aimed to derive a water-soluble antimicrobial agent for food from 4-methylthio-3-butenyl isothiocyanate (MTBI), by reaction with amino acids. First, it was confirmed that MTBI was mostly degraded on an agar plate during incubation for 6-8 hours, suggesting that MTBI probably produced antimicrobial product(s) during the degradation. Consequently, antimicrobial activity of the water-soluble degradation products was studied. The water-soluble fraction showed remarkable growth-inhibition on some fungi and Gram-positive bacteria, and the active component was identified as 2-thioxo-3-pyrrolidinecarbaldehyde(TPC). On the other hand, formation of some compounds including ninhydrin-positive, non-fluorescent and brightly yellow substances were observed by the reaction of MTBI with amino acids. Since the reaction products exhibited a prominent growth-inhibition on yeasts and fungi, chromatographic separation of the active component(s) was tried by using reverse phase silica gel(ODS). The antimicrobial activity was monitored by disk-diffusion method using Eurotium chevarieri and Cladosporium colocasiae. The activity was mostly found in the fractions eluted with 10% methanol from ODS-column. Thus, the active fraction was further separated on a ODS-thin layr plate. Both lower(Rf 0-0.25) and higher(Rf 0.45-0.70) Rf-fractions contained antifungal components, of which the former fraction had more stronger activity. It was presumed that the active component in the lower Rf-fraction was attributable to the brightly yellow component formed by the reaction of MTBI with amino acids. In the higher Rf-fraction, the antifungal component, TPC was detected together with both the ninhydrin-positive and non-fluorescent components. From the results, it was clarified that separation of the antimicrobial products formed by reaction of MTBI with amino acids is attainable by the reverse phase silica gel chromatography. Identification of the antimicrobial products was remained to be furt

本项目的目的是从4-甲硫基-3-丁烯基异硫氰酸酯（MTBI）中，通过与氨基酸反应，得到一种水溶性食品抗菌剂。首先，证实MTBI在培养6-8小时期间在琼脂平板上大部分降解，表明MTBI在降解期间可能产生抗微生物产物。因此，研究了水溶性降解产物的抗微生物活性。水溶性部分对几种真菌和革兰氏阳性菌有明显的生长抑制作用，其活性成分经鉴定为2-硫代-3-吡咯烷甲醛（TPC）。另一方面，通过MTBI与氨基酸的反应，观察到一些化合物的形成，包括茚三酮阳性、非荧光和亮黄色物质。由于反应产物对酵母和真菌表现出显著的生长抑制，因此通过使用反相硅胶（ODS）尝试了活性组分的色谱分离。采用纸片扩散法测定其抑菌活性。ODS柱上用10%甲醇洗脱的组分中活性最高。因此，在ODS-薄层板上进一步分离活性级分。Rf值较低（0-0.25）和较高（0.45-0.70）的组分均含有抑菌活性成分，其中Rf值较高的组分抑菌活性较强。据推测，较低Rf级分中的活性组分归因于MTBI与氨基酸反应形成的亮黄色组分。在较高的Rf-级分，抗真菌成分，TPC一起检测茚三酮阳性和非荧光成分。从结果中可以清楚地看出，通过反相硅胶色谱法可以分离MTBI与氨基酸反应形成的抗微生物产物。抗菌药物的鉴别仍有待进一步研究

项目成果

Y.Uda,H.Matsuoka,H.Kumagami,H.Shima and Y.Maeda: "Stability and Antimicrobial Property of 4-Methylthio-3-butenyl Isothiocyanate,the Pungent Principle in Radish" 日本食品工業学会誌. 40. 743-746 (1993)

Y.Uda、H.Matsuoka、H.Kumagami、H.Shima 和 Y.Maeda：“4-甲硫基-3-丁烯基异硫氰酸酯的稳定性和抗菌特性，萝卜中的辛辣原理”日本食品工业协会杂志 40。 743 -746 (1993)

Y.Uda,H.Matsuoka,H.Shima,H.Kumagami and Y.Maeda: "Antimicrobial Activity of Water-soluble Products Derived from Radish Mustard Oil and Identification of an Active Component therein" 日本食品工業学会誌. 40. 801-806 (1993)

Y.Uda、H.Matsuoka、H.Shima、H.Kumagami 和 Y.Maeda：“萝卜芥末油水溶性产品的抗菌活性及其活性成分的鉴定”日本食品工业协会杂志 40。 801 -806 (1993)