Polyphosphate kinase (PPK) is important for industrial processes involving ATP regeneration. While a variety of methods have been reported for regenerating ATP from ADP, few have explored enzyme catalyzed ATP regeneration from cheaper and stable AMP. In this work, PPKs from different sources were expressed and their catalytic activity were tested at different reaction temperatures, reaction pH and with different polyphosphate (polyP_n) types. The ATP regeneration system for glutathione (GSH) synthesis was established using a single PPK capable of phosphorylating AMP to synthesize ATP from AMP and short chain polyP_n. GSH yield was obtained using adenosine mono-, di- and triphosphates, which confirmed the flexibility of our constructed ATP regeneration system coupled with GSH synthesis via bifunctional GSH synthase. Finally, optimization of the GSH synthesis yielded conversion value above 80 %. Overall, these results illustrate that PPK is suitable for a broader range of substrates than previously expected, and has great untapped potential for applications involving ATP regeneration.

多磷酸激酶 (PPK) 对于涉及 ATP 再生的工业过程很重要。虽然已经报道了从 ADP 再生 ATP 的各种方法，但很少有人探索酶催化从更便宜和稳定的 AMP 再生 ATP。在这项工作中，表达了来自不同来源的 PPK，并在不同的反应温度、反应 pH 值和不同的多磷酸盐 (polyP _n ) 类型下测试了它们的催化活性。谷胱甘肽 (GSH) 合成的 ATP 再生系统是使用能够磷酸化 AMP 的单个 PPK 从 AMP 和短链 polyP _n合成 ATP 建立的. 使用单磷酸腺苷、二磷酸腺苷和三磷酸腺苷获得 GSH 产量，这证实了我们构建的 ATP 再生系统与通过双功能 GSH 合酶合成 GSH 的灵活性。最后，GSH 合成的优化产生了 80% 以上的转化值。总体而言，这些结果表明 PPK 适用于比以前预期的更广泛的底物，并且在涉及 ATP 再生的应用方面具有巨大的未开发潜力。