Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis.
Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis.
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DOI：
10.1093/femsle/fnv180
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发表时间：
2015-11
影响因子：
2.1
通讯作者：
Andrij Fedashchin;William H. Cernota;Melissa C. Gonzalez;Benjamin I. Leach;Noelle Kwan;R. Wesley;J. Weber
中科院分区：
文献类型：
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作者：
Andrij Fedashchin;William H. Cernota;Melissa C. Gonzalez;Benjamin I. Leach;Noelle Kwan;R. Wesley;J. Weber
对红霉素糖多孢子菌b进行了单周期的菌株改良，发现了15种影响红霉素产量的基因型。转座子诱变产生的基因型在筛选中出现的频率约为3%。发现了影响中枢代谢和调控基因的突变，以及水解酶、肽酶、糖基转移酶和未知基因的突变。当从微琼脂塞发酵放大到摇瓶时，只有一个突变体保留了高红霉素产量。该突变体敲除了编码细胞壁相关水解酶的cwh1基因（SACE_1598）。cwh1基因敲除在固体培养基上产生明显的生长和形态缺陷。该研究表明，随机转座子突变揭示了菌株改良相关基因，可能对菌株工程有用。
A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ~3%. Mutations affecting central metabolism and regulatory genes were found, as well as hydrolases, peptidases, glycosyl transferases and unknown genes. Only one mutant retained high erythromycin production when scaled-up from micro-agar plug fermentations to shake flasks. This mutant had a knockout of the cwh1 gene (SACE_1598), encoding a cell-wall-associated hydrolase. The cwh1 knockout produced visible growth and morphological defects on solid medium. This study demonstrated that random transposon mutagenesis uncovers strain improvement-related genes potentially useful for strain engineering.