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通过构建pya26的同框缺失突变株以及回补菌株,对其发酵产物进行高效液相色谱和质谱分析,鉴定醛基转移酶Pya26在远端霉素(distamycin)和Compound 3(disgocidine)生物合成过程中的功能.结果显示:pya26同框缺失突变株丧失了远端霉素和disgocidine的产生能力;Δpya26回补菌株仅恢复了远端霉素的生产能力,但产量很低;而酰胺水解酶编码基因pya25与pya26的双基因回补菌株不仅恢复了远端霉素和disgocidine的生产能力,且与单基因回补菌株相比产量有明显增加.结果表明:pya26是远端霉素以及disgocidine生物合成的必需基因,Pya25与Pya26协同完成远端霉素和disgocidine的生物合成.
By constructing the in-frame deletion mutants of pya26 and the complementing strains, their fermentation products were identified by high performance liquid chromatography and mass spectrometry, and the biosynthesis of distycinin and compound 3 (disgocidine) by aldehydetransferase Pya26 was identified The results showed that: pya26 in-frame deletion mutant lost the ability of farmycin and disgocidine production; Δpya26 back-up strain restored only the production of farmycin, but the yield was low; while the amidolytic enzyme coding The double-gene refolding strains of genes pya25 and pya26 not only restored the production ability of farmycin and disgocidine, but also significantly increased the yield compared with the single-gene covering strains.The results showed that pya26 was far-namycin and disgocidine biosynthesis The essential genes, Pya25 and Pya26, work together to accomplish the biosynthesis of farmycin and disgocidine.