一株吡咯喹啉醌产生菌的筛选和初步鉴定

Isolation and Preliminary Identification of a Pyrroloquinoline Quinone Producing Strain

目的:筛选新的吡咯喹啉醌(PQQ)产生菌株。方法:收集土壤样品富集培养,分离菌株,NBT染色法快速筛选PQQ产生菌,经Native-PAGE复筛,NBT-Gly法和分光光度法相结合测定菌株PQQ产量;选取产量最高的1株菌考察菌株生长条件,扩增16S rDNA并测序,Blast比对分析确定菌株分类位置。结果:从500余份土壤样品中筛选得到1株PQQ产生菌T28,PQQ产量达13.3 mg/L。该菌株16S rDNA序列与生丝微菌的同源性为96%,确定了T28的最适生长培养基配方。结论:从土壤中筛选到1株新的PQQ产生菌,命名为生丝微菌T28。

Objective： To isolate new pyrroloquinoline quinone（PQQ） producing strains. Methods： The PQQ pro- ducing strains were enriched from the collected soil samples, and were isolated by NBT staining test and further screened by Native-PAGE. The PQQ yield was determined by NBT-glycine method and spectrophotometry. The strain with the highest yield was selected to investigate the growth conditions. The location of this strain was determined by blasting alignment analysis of its 16S rDNA sequence. Results： From about 500 soil samples, one PQQ producing strain T28 with 13.3 mg/L PQQ yield was isolated. The 16S rDNA sequence of this strain shared 96% homology with Hyphomicrobium, and the T28 optimal growth medium formulation was determined. Conclusion： A new Hyphomicrobium T28 strain of producing PQQ was isolated from the soil.