The purification and characterization of PGM (Phosphoglucomutase) from Cordyceps militaris ( C. militaris ) was investigated. PGM was purified using a combination of ultrafiltration, salting-out and ion exchange chromatography resulting in 4.23-fold enhancement of activity with a recovery of 20.01%. Molecular mass was 50.01 kDa by SDS-PAGE. The optimal activity was achieved at pH 7.5 and 30 °C with NADPH as substrate. The results showed that SDS, DTT Li + , Cu 2+ , Na + , Mn 2+ and Al 3+ were effective PGM inhibitors; whereas glycerol, Zn 2+ , Mg 2+ , Ca 2+ , Fe 2+ and Fe 3+ could enhance the activity of PGM, and the K m and V max values were 11.62 mmol/L and 416.67 U/mL, respectively. At the same time, qRT-PCR was used to test the changes of mRNA transcription level of PGM gene encoding under two fermentation conditions: basic medium and optimized medium. The relative quantitative results of PGM target genes resulting in 2.60-fold enhancement than the control group.

中文翻译：

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蛹虫草磷酸葡萄糖变位酶基因表达的酶学表征和验证

研究了来自蛹虫草 (C. militaris) 的 PGM (磷酸葡萄糖变位酶) 的纯化和表征。PGM 使用超滤、盐析和离子交换色谱的组合进行纯化，导致活性提高 4.23 倍，回收率为 20.01%。通过SDS-PAGE，分子量为50.01 kDa。最佳活性在 pH 7.5 和 30 °C 下以 NADPH 为底物实现。结果表明，SDS、DTT Li + 、Cu 2+ 、Na + 、Mn 2+ 和Al 3+ 是有效的PGM抑制剂；而甘油、Zn 2+ 、Mg 2+ 、Ca 2+ 、Fe 2+ 和Fe 3+ 可以增强PGM的活性，K m 和V max 值分别为11.62 mmol/L和416.67 U/mL。同时采用qRT-PCR检测两种发酵条件下编码PGM基因mRNA转录水平的变化：基础培养基和优化培养基。PGM 靶基因的相对定量结果导致比对照组提高 2.60 倍。