Establishment and Optimization of the High-Throughput Measurement of Pyrroloquinoline Quinone

Abstract:

Pyrroloquinoline quinone（PQQ） is a new coenzyme that has broad application prospect. Previous methods used for measuring PQQ in fermentation broth are defective and low-efficient. In this study，D-glucose dehydrogenase（PQQGDH） from Escherichia coli K-12 was expressed in E. coli BL21 and then purified by the nickel-affinity chromatography column. For the high-throughput measurement of PQQ the optimal reaction system was found by orthogonal experiment to contain 30 μL PQQGDH，2 μL PQQ sample and 1.2 mL chromogenic reagent. The method was linear within a wide range and had high precision and good reproducibility. Compared with traditional methods，the high-throughput measurement of PQQ using purified PQQGDH，96-well plate and microplate reader are herein more accurate and efficient.