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糖和氨基酸对出芽短梗霉 NRRL 58536 抗四种曲霉属真菌活性的影响。

Effects of sugar and amino acid supplementation on Aureobasidium pullulans NRRL 58536 antifungal activity against four Aspergillus species.

机构信息

Plant Biomass Utilization Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.

出版信息

Appl Microbiol Biotechnol. 2013 Sep;97(17):7821-30. doi: 10.1007/s00253-013-5069-5. Epub 2013 Jul 5.

Abstract

Cultured cell extracts from ten tropical strains of Aureobasidium pullulans were screened for antifungal activity against four pathogenic Aspergillus species (Aspergillus flavus, Aspergillus niger, Aspergillus fumigatus, and Aspergillus terreus) using the well diffusion and conidial germination inhibition assays. The crude cell extract from A. pullulans NRRL 58536 resulted in the greatest fungicidal activity against all four Aspergillus species and so was selected for further investigation into enhancing the production of antifungal activity through optimization of the culture medium, carbon source (sucrose and glucose) and amino acid (phenylalanine, proline, and leucine) supplementation. Sucrose did not support the production of any detectable antifungal activity, while glucose did with the greatest antifungal activity against all four Aspergillus species being produced in cells grown in medium containing 2.5 % (w/v) glucose. With respect to the amino acid supplements, variable trends between the different Aspergillus species and amino acid combinations were observed, with the greatest antifungal activities being obtained when grown with phenylalanine plus leucine supplementation for activity against A. flavus, proline plus leucine for A. terreus, and phenylalanine plus proline and leucine for A. niger and A. fumigatus. Thin layer chromatography, spectrophotometry, high-performance liquid chromatography, (1)H-nuclear magnetic resonance, and MALDI-TOF mass spectrometry analyses were all consistent with the main component of the A. pullulans NRRL 58536 extracts being aureobasidins.

摘要

从十种热带地区的出芽短梗霉(Aureobasidium pullulans)菌株的培养细胞提取物中筛选出对四种致病性曲霉属物种(黄曲霉(Aspergillus flavus)、黑曲霉(Aspergillus niger)、烟曲霉(Aspergillus fumigatus)和构巢曲霉(Aspergillus terreus))具有抗真菌活性的物质,使用平板扩散和孢子萌发抑制试验进行测定。A. pullulans NRRL 58536 的粗细胞提取物对所有四种曲霉属物种均具有最大的杀菌活性,因此被选择用于进一步研究通过优化培养基、碳源(蔗糖和葡萄糖)和氨基酸(苯丙氨酸、脯氨酸和亮氨酸)添加来提高抗真菌活性的生产。蔗糖不支持任何可检测的抗真菌活性的产生,而葡萄糖支持产生最大的抗真菌活性,所有四种曲霉属物种在含有 2.5%(w/v)葡萄糖的培养基中生长的细胞中产生。就氨基酸补充而言,观察到不同的曲霉属物种和氨基酸组合之间的趋势不同,当与苯丙氨酸加亮氨酸补充生长时,对黄曲霉的抗真菌活性最大,对构巢曲霉的抗真菌活性最大,对黑曲霉和烟曲霉的抗真菌活性最大与脯氨酸加亮氨酸一起生长,与苯丙氨酸加脯氨酸和亮氨酸一起生长。薄层层析、分光光度法、高效液相色谱法、(1)H 核磁共振和 MALDI-TOF 质谱分析均一致表明,A. pullulans NRRL 58536 提取物的主要成分是 Aureobasidins。

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