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Comparative genomic and proteomic analyses of Clostridium acetobutylicum Rh8 and its parent strain DSM 1731 revealed new understandings on butanol tolerance

机译:丙酮丁醇梭菌Rh8及其亲本菌株DSM 1731的基因组和蛋白质组学比较分析揭示了对丁醇耐受性的新认识

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Clostridium acetobutylicum strain Rh8 is a butanol-tolerant mutant which can tolerate up to 19 g/L butanol, 46% higher than that of its parent strain DSM 1731. We previously performed comparative cytoplasm- and membrane-proteomic analyses to understand the mechanism underlying the improved butanol tolerance of strain Rh8. In this work, we further extended this comparison to the genomic level. Compared with the genome of the parent strain DSM 1731, two insertion sites, four deletion sites, and 67 single nucleotide variations (SNVs) are distributed throughout the genome of strain Rh8. Among the 67 SNVs, 16 SNVs are located in the predicted promoters and intergenic regions; while 29 SNVs are located in the coding sequence, affecting a total of 21 proteins involved in transport, cell structure, DNA replication, and protein translation. The remaining 22 SNVs are located in the ribosomal genes, affecting a total of 12 rRNA genes in different operons. Analysis of previous comparative proteomic data indicated that none of the differentially expressed proteins have mutations in its corresponding genes. Rchange Algorithms analysis indicated that the mutations occurred in the ribosomal genes might change the ribosome RNA thermodynamic characteristics, thus affect the translation strength of these proteins. Take together, the improved butanol tolerance of C. acetobutylicum strain Rh8 might be acquired through regulating the translational process to achieve different expression strength of genes involved in butanol tolerance.
机译:丙酮丁醇梭菌菌株Rh8是耐丁醇的突变体,可耐受高达19 g / L的丁醇,比其亲本菌株DSM 1731高46%。提高了Rh8菌株的丁醇耐受性。在这项工作中,我们进一步将此比较扩展到了基因组水平。与亲本菌株DSM 1731的基因组相比,在Rh8菌株的整个基因组中分布了两个插入位点,四个缺失位点和67个单核苷酸变异(SNV)。在67个SNV中,有16个SNV位于预测的启动子和基因间区域。而29个SNV位于编码序列中,总共影响21种涉及运输,细胞结构,DNA复制和蛋白质翻译的蛋白质。其余22个SNV位于核糖体基因中,影响不同操纵子中的总共12个rRNA基因。对先前比较蛋白质组学数据的分析表明,差异表达的蛋白质均未在其相应基因中具有突变。 Rchange算法分析表明,核糖体基因中发生的突变可能会改变核糖体RNA的热力学特性,从而影响这些蛋白质的翻译强度。综上所述,可以通过调节翻译过程以获得参与丁醇耐受性的基因的不同表达强度来获得提高的丙酮丁醇梭菌菌株Rh8的丁醇耐受性。

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