吡咯酰胺类化合物合成途径中醛基转移酶Pya26的功能研究

Functional Identification of Pya26,a Formyltransferase Involved in the Biosynthesis of Pyrrole Amide Compound

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.

The culture extract of the pya26 in-frame deletion and complementation mutants was detected by HPLC and LC-MS to identify the function of Pya26, a formyltransferase involved in the biosynthesis of distamycin and disgosidine. The results showed that the production of distamycin and disgocidine was abolished in pya26 in-frame de- letion mutant. While APya 26 complementation mutant only partially restored the production of distamycin, as pya 25- pya 26 co-eomplementation mutant produced much higher level of distamyein and disgocidine than that of the △pya 26 complementation mutant. The conclusion was that pya26 is essential for the biosynthesis of distamycin and disgoci- dine, and Pya25 and Pya26 function in a coordinated manner in the biosynthesis of distamycin and disgocidine.