Arthrobacter sp. CGMCC 3584 is used for the industrial production of cyclic adenosine monophosphate (cAMP). However, because of the paucity of genetic engineering tools for genetic manipulation on Arthrobacter species, only a few metabolically engineered Arthrobacter have been constructed and investigated. In this study, for the first time, we constructed an arpde knockout mutant of Arthrobacter without any antibiotic resistance marker by a PCR-targeting-based homologous recombination method. Our results revealed that the deletion of arpde had little effect on biomass production and improved cAMP production by 31.1%. Furthermore, we compared the transcriptomes of the arpde knockout strain and the wild strain, aiming to understand the capacities of cAMP production due to arpde inactivation at the molecular level. Comparative transcriptomic analysis revealed that arpde inactivation had two major effects on metabolism: inhibition of glycolysis, PP pathway, and amino acid metabolism (phenylalanine, tryptophan, branched-chain amino acids, and glutamate metabolism); promotion of the purine metabolism and carbon flux from the precursor 5'-phosphoribosyl 1-pyrophosphate, which benefited cAMP production.

中文翻译：

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关节杆菌sp中pde基因的敲除。CGMCC 3584及其对cAMP产生的影响的转录组分析。

关节杆菌 CGMCC 3584用于工业生产环状单磷酸腺苷（cAMP）。但是，由于缺乏对节杆菌属物种进行遗传操作的基因工程工具，因此仅构建和研究了少数经代谢工程改造的节杆菌。在这项研究中，我们首次通过基于PCR靶向的同源重组方法构建了无任何抗生素抗性标记的节杆菌的arpde基因敲除突变体。我们的研究结果表明删除arpde对生物量生产影响不大，并将cAMP产量提高了31.1％。此外，我们比较了阿尔普基因敲除菌株和野生菌株的转录组，旨在了解由于在分子水平上的阿尔普分子失活而产生的cAMP的能力。对比转录组分析显示，arpde失活对代谢有两个主要影响：抑制糖酵解，PP途径和氨基酸代谢（苯丙氨酸，色氨酸，支链氨基酸和谷氨酸代谢）；促进嘌呤代谢和前体5'-磷酸核糖基1-焦磷酸的碳通量，这有利于cAMP的生产。