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首页> 外文期刊>Applied Microbiology and Biotechnology >Tryptophan promotes morphological and physiological differentiation in Streptomyces coelicolor
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Tryptophan promotes morphological and physiological differentiation in Streptomyces coelicolor

机译：色氨酸促进天蓝色链霉菌的形态和生理分化

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摘要

The molecular mechanisms regulating tryptophan biosynthesis in actinomycetes are poorly understood; similarly, the possible roles of tryptophan in the differentiation program of microorganism life-cycle are still underexplored. To unveil the possible regulatory effect of this amino acid on gene expression, an integrated study based on quantitative teverse transcription-PCR (qRT-PCR) and proteomic approaches was performed on the actinomycete model Streptomyces coelicolor. Comparative analyses on the microorganism growth in a minimal medium with or without tryptophan supplementation showed that biosynthetic trp gene expression in S. coelicolor is not subjected to a negative regulation by the presence of the end product. Conversely, tryptophan specifically induces the transcription of trp genes present in the biosynthetic gene cluster of the calcium-dependent antibiotic (CDA), a lipopeptide containing d- and l-tryptophan residues. In addition, tryptophan stimulates the transcription of the CDA gene cluster regulator cdaR and, coherently, CDA production. Surprisingly, tryptophan also promotes the production of actinorhodin, another antibiotic that does not contain this amino acid in its structure. Combined 2D-DIGE and nano liquid chromatography electrospray linear ion trap tandem mass spectrometry (LC-ESI-LIT-MS/MS) analyses revealed that tryptophan exerts a growth-stage-dependent global effect on S. coelicolor proteome, stimulating anabolic pathways and promoting the accumulation of key factors associated with morphological and physiological differentiation at the late growth stages. Phenotypic observations by scanning electron microscopy and spore production assays demonstrated an increased sporulation in the presence of tryptophan. Transcriptional analysis of catabolic genes kynA and kynB suggested that the actinomycete also uses tryptophan as a carbon and nitrogen source. In conclusion, this study originally provides the molecular basis underlying the stimulatory effect of tryptophan on the production of antibiotics and morphological development program of this actinomycete.
机译：对放线菌中色氨酸生物合成的分子机制了解甚少。同样，色氨酸在微生物生命周期分化程序中的可能作用仍未得到充分研究。为了揭示该氨基酸可能对基因表达的调控作用，在放线菌模型天蓝色链霉菌（Streptomyces coelicolor）上进行了基于定量逆转录PCR（qRT-PCR）和蛋白质组学方法的综合研究。在添加或不添加色氨酸的基本培养基中对微生物生长的比较分析表明，终产物的存在不会对天蓝色链霉菌中生物合成的trp基因表达产生负面影响。相反，色氨酸特异性地诱导存在于钙依赖性抗生素（CDA）的生物合成基因簇中的trp基因的转录，钙依赖性抗生素是一种含有D和L色氨酸残基的脂肽。此外，色氨酸刺激CDA基因簇调节剂cdaR的转录，并相应地刺激CDA的产生。出乎意料的是，色氨酸还促进肌动蛋白的生成，肌动蛋白是另一种在其结构中不含该氨基酸的抗生素。结合2D-DIGE和纳米液相色谱电喷雾线性离子阱串联质谱（LC-ESI-LIT-MS / MS）分析，色氨酸对Coelicolor蛋白质组蛋白具有生长阶段依赖性的整体效应，刺激合成代谢途径并促进生长后期与形态和生理分化有关的关键因素的积累。通过扫描电子显微镜和孢子产生测定的表型观察表明在色氨酸存在下孢子形成增加。分解代谢基因kynA和kynB的转录分析表明，放线菌也使用色氨酸作为碳和氮源。总之，本研究最初提供了色氨酸对这种放线菌的抗生素产生和形态发展程序的刺激作用的分子基础。

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