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Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis

机译:Saccharopolyspora erythraea基因组的随机转座子诱变揭示了影响红霉素生物合成的其他基因

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摘要

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.
机译:在糖多孢菌mutB中进行了单株菌株改良,发现了15种影响红霉素生产的基因型。转座子诱变产生的基因型以约3%的频率出现在筛选中。发现影响中央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。当从微琼脂塞发酵扩大到摇瓶时,只有一个突变体保留了高红霉素生产。此突变体具有编码细胞壁相关水解酶的cwh1基因(SACE_1598)的敲除。 cwh1基因敲除在固体培养基上产生可见的生长和形态缺陷。这项研究表明,随机转座子诱变发现了与菌株改良相关的基因,可能对菌株工程有用。

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