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从酸面团植物乳杆菌21B菌株中分离新型抗真菌化合物并进行表征

Purification and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B.

作者信息

Lavermicocca P, Valerio F, Evidente A, Lazzaroni S, Corsetti A, Gobbetti M

机构信息

Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, 70125 Bari, Italy.

出版信息

Appl Environ Microbiol. 2000 Sep;66(9):4084-90. doi: 10.1128/AEM.66.9.4084-4090.2000.

Abstract

Sourdough lactic acid bacteria were selected for antifungal activity by a conidial germination assay. The 10-fold-concentrated culture filtrate of Lactobacillus plantarum 21B grown in wheat flour hydrolysate almost completely inhibited Eurotium repens IBT18000, Eurotium rubrum FTDC3228, Penicillium corylophilum IBT6978, Penicillium roqueforti IBT18687, Penicillium expansum IDM/FS2, Endomyces fibuliger IBT605 and IDM3812, Aspergillus niger FTDC3227 and IDM1, Aspergillus flavus FTDC3226, Monilia sitophila IDM/FS5, and Fusarium graminearum IDM623. The nonconcentrated culture filtrate of L. plantarum 21B grown in whole wheat flour hydrolysate had similar inhibitory activity. The activity was fungicidal. Calcium propionate at 3 mg ml(-1) was not effective under the same assay conditions, while sodium benzoate caused inhibition similar to L. plantarum 21B. After extraction with ethyl acetate, preparative silica gel thin-layer chromatography, and chromatographic and spectroscopic analyses, novel antifungal compounds such as phenyllactic and 4-hydroxy-phenyllactic acids were identified in the culture filtrate of L. plantarum 21B. Phenyllactic acid was contained at the highest concentration in the bacterial culture filtrate and had the highest activity. It inhibited all the fungi tested at a concentration of 50 mg ml(-1) except for P. roqueforti IBT18687 and P. corylophilum IBT6978 (inhibitory concentration, 166 mg ml(-1)). L. plantarum 20B, which showed high antimold activity, was also selected. Preliminary studies showed that phenyllactic and 4-hydroxy-phenyllactic acids were also contained in the bacterial culture filtrate of strain 20B. Growth of A. niger FTDC3227 occurred after 2 days in breads started with Saccharomyces cerevisiae 141 alone or with S. cerevisiae and Lactobacillus brevis 1D, an unselected but acidifying lactic acid bacterium, while the onset of fungal growth was delayed for 7 days in bread started with S. cerevisiae and selected L. plantarum 21B.

摘要

通过分生孢子萌发试验筛选具有抗真菌活性的酸面团乳酸菌。在小麦粉水解物中培养的植物乳杆菌21B的10倍浓缩培养滤液几乎完全抑制了间型曲霉IBT18000、红曲霉FTDC3228、嗜棒青霉IBT6978、罗克福特青霉IBT18687、扩展青霉IDM/FS2、纤维内孢霉IBT605和IDM3812、黑曲霉FTDC3227和IDM1、黄曲霉FTDC3226、嗜热毁丝霉IDM/FS5以及禾谷镰刀菌IDM623。在全麦粉水解物中培养的植物乳杆菌21B的未浓缩培养滤液具有相似的抑制活性。该活性具有杀真菌作用。在相同试验条件下,3 mg/ml的丙酸钙无效,而苯甲酸钠引起的抑制作用与植物乳杆菌21B相似。经乙酸乙酯萃取、制备硅胶薄层层析以及色谱和光谱分析后,在植物乳杆菌21B的培养滤液中鉴定出了新型抗真菌化合物,如苯乳酸和4-羟基苯乳酸。苯乳酸在细菌培养滤液中的浓度最高,活性也最高。它在浓度为50 mg/ml时抑制了所有测试的真菌,但罗克福特青霉IBT18687和嗜棒青霉IBT6978除外(抑制浓度为166 mg/ml)。还筛选出了具有高抗霉菌活性的植物乳杆菌20B。初步研究表明,菌株20B的细菌培养滤液中也含有苯乳酸和4-羟基苯乳酸。单独使用酿酒酵母141或与酿酒酵母和短乳杆菌1D(一种未筛选但能酸化的乳酸菌)制作面包时,黑曲霉FTDC3227在2天后开始生长,而在以酿酒酵母和筛选出的植物乳杆菌21B制作的面包中,真菌生长的起始时间延迟了7天。

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