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首页> 外文期刊>Catalysis science & technology >Design of an in vitro multienzyme cascade system for the biosynthesis of nicotinamide mononucleotide
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Design of an in vitro multienzyme cascade system for the biosynthesis of nicotinamide mononucleotide

机译：烟酰胺单核苷酸生物合成的体外多酶级联系统的设计

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摘要

For the biosynthesis of nicotinamide mononucleotide (NMN), three artificial pathways including the nicotinamide ribose phosphorylation pathway, adenosine phosphate pyrophosphorylation pathway, and adenosine phosphate hydrolysis (APH) pathway were designed and successfully conducted to produce NMN in vitro. The APH pathway, using AMP nucleosidase, ribose-phosphate diphosphokinase, and nicotinamide phosphoribosyltransferase (NAMPT), exhibited the highest level of NMN synthesis. To further improve NMN production via the APH pathway, various NAMPT orthologues were screened. The effects of temperature, pH, metal ions and enzyme ratios were further systematically investigated, and the accumulation of ADP was identified limiting pathway efficiency. Subsequently, an ATP recycling process was achieved by adding polyphosphate kinase 2 to convert ADP to ATP. With the optimized four multienzyme cascade catalysis systems, the NMN titer was increased to 9 mmol L−1 (3.0 g L−1) from 600 μmol L−1 (0.2 g L−1) in vitro. This is the first study to use a multienzyme cascade catalysis process for NMN biosynthesis.
机译：对于烟酰胺单核苷酸（NMN）的生物合成，包括烟酰胺核糖磷酸化途径，包括烟酰胺核糖磷酸化途径，磷酸腺苷磷酸磷酸化途径，以及磷酸腺苷水解（APH）途径的生产并成功地产生NMN。使用AMP核苷酶，核糖 - 磷酸二磷酸激酶和烟酰胺磷酸贝糖基转移酶（NAMPT）的APH途径表现出最高水平的NMN合成水平。为了通过APH途径进一步改善NMN的生产，筛选了各种NAMPT直系同源物。进一步研究了温度，pH值，金属离子和酶比的影响，并确定了ADP的积累限制途径效率。随后，通过添加多磷酸激酶2将ADP转换为ATP来实现ATP回收过程。通过优化的四个多元素级联催化系统，从600μmolL -1（0.2 g L -1）体外，NMN滴度增加到9 mmol L -1（3.0 g L -1）。这是第一个使用多元素级联催化过程进行NMN生物合成的研究。

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