吡咯喹啉醌(Pyrroloquinoline quinone,PQQ)作为一种新型的氧化还原酶辅酶,在医药和食品等领域有广阔的应用前景。为改善扭脱甲基杆菌Methylobacterium extorquens AM1 PQQ生产性能,采用常压室温等离子体(Atmospheric and room temperature plasma,ARTP)进行诱变,结合高通量快速筛选方法,得到以PQQ产量为指标的正向突变株。ARTP诱变的菌株正突变率为31.6%,筛选得到的较优正突变株M.extorquens AM1(E-F3),PQQ产量达到54.0 mg/L,是出发菌株的近3倍。系统的高通量方法筛选ARTP诱变菌为后续进一步提高M.extorquens AM1菌株PQQ的产量奠定了基础,亦为改善菌株生产性能提供了新思路。
As a novel cofactor of oxidoreductase,pyrroloquinoline quinone(PQQ) has a great potential of application in medicine,food industries.In order to improve the efficiency of the PQQ production by Methylobacterium extorquens AM1,the strain was treated by atmospheric and room temperature plasma(ARTP).Positive mutants with changes in PQQ yield were obtained based on a high-throughput screening approach.After ARTP treatment,analysis data show that the positive mutation rate was 31.6%.Furthermore,we obtained an excellent positive mutant M.extorquens AM1(E-F3) with the yield of 54.0 mg/L PQQ,which was approximately 3 times as much compared with that of the wild-type strain.The robust high-throughput screening method for mutagenesis by ARTP improves PQQ production.In addition,this method also provides a new strategy for further strain improvement.