Optimisation of aurodox production by Streptomyces goldiniensis.
Optimisation of aurodox production by Streptomyces goldiniensis.
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金链霉菌 Aurodox 生产的优化。
DOI：
10.1099/acmi.0.000572.v1
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
E McHugh R
中科院分区：
文献类型：
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作者：
E McHugh R
Aurodox是戈尔丁链霉菌的一种专门代谢产物，最初被鉴定具有抗链球菌和抗葡萄球菌活性。最近，人们重新关注对aurodox的研究，因为该化合物已被证明能抑制肠致病性和肠出血性大肠杆菌（EPEC/EHEC）的毒力。Aurodox能够通过下调III型分泌系统（这些病原体中的主要毒力因子）来抑制EPEC和EHEC感染中的宿主定植。为了进一步研究活性，需要增加化合物的量以进行深入的作用机制研究。为了确定合适的aurodox生产条件，测试了许多相关的发酵培养基以提高aurodox产率，然后进行生物测定和液相色谱质谱分析。葡萄糖酵母提取物培养基（GYM）被确定为检测的主要生产培养基。对S. goldiniensis在GYM培养基中的生长建立了产金霉素的最佳时间范围。在S. goldiniensis液体GYM培养基发酵。为了提高总的aurodox滴度，采取异源表达（HE）方法，并研究了四种潜在的HE宿主。这些数据表明，aurodox生物合成基因簇可以在两个S. collinus Tü 365和S. coelicolor M1152与S. goldiniensis。这项工作展示了如何将传统的生理分析和现代遗传技术相结合，以改善天然产物的生产，以进行详细的作用模式研究。
Aurodox is a specialised metabolite of Streptomyces goldiniensis originally identified for its anti-streptococcal and anti-staphylococcal activity. More recently, there has been a renewed focus on the study of aurodox as the compound has been shown to inhibit virulence in both Enteropathogenic and Enterohemorrhagic Escherichia coli (EPEC/EHEC). Aurodox is able to inhibit host colonization in EPEC and EHEC infections via the downregulation of the Type III Secretion System – a major virulence factor in these pathogens. To further investigate activity, increased quantities of the compound are required for in-depth mechanism of action studies. To identify suitable aurodox production conditions, a number of relevant fermentation media were tested for enhanced aurodox yield, followed by bioassay and Liquid Chromatography Mass Spectrometry analysis. Glucose Yeast Extract medium (GYM) was identified as the leading production medium tested. Further analysis of S. goldiniensis growth in GYM medium established the optimum temporal range for aurodox production. A minimum growth period of 154 hours is required for detectable aurodox production in S. goldiniensis fermentations in liquid GYM medium. To improve overall aurodox titres, a heterologous expression (HE) approach was taken and four potential HE hosts were investigated. These data indicated that the aurodox biosynthetic gene cluster can be heterologously expressed in both S. collinus Tü365 and S. coelicolor M1152, with increased aurodox titres compared to those of S. goldiniensis. This work demonstrates how traditional physiological analysis and modern genetic techniques can be combined to improve natural product production for detailed mode-of-action studies.