Knockout of pde gene in Arthrobacter sp. CGMCC 3584 and transcriptomic analysis of its effects on cAMP production
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.
KeywordsArthrobacter Knockout of arpde gene Transcriptome cAMP fermentation
This work was supported by the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture (Grant No. XTE1842), the young investigator grant program of National Natural Science Foundation of China (Grant No. 21706123), the National Natural Science Foundation of China, General Program (Grant No. 31972503), the key program of the National Natural Science Foundation of China (Grant No. 21636003), Jiangsu Natural Science Fund for Distinguished Young Scholars (Grant No. BK20190035), the Program for Changjiang Scholars and Innovative Research Team in University (Grant No. IRT_14R28), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
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Conflict of interest
The authors declare that they have no conflict of interest.
The article does not contain any researches with human participants and/or animals performed by any of the authors.
Human and animal rights
The research did not involve human participants and/or animals.
- 5.Niewerth H, Parschat K, Rauschenberg M, Ravoo BJ, Fetzner S (2013) The PaaX-type repressor MeqR2 of Arthrobacter sp. strain Rue61a, involved in the regulation of quinaldine catabolism, binds to its own promoter and to catabolic promoters and specifically responds to anthraniloyl coenzyme A. J Bacteriol 195(5):1068–1080PubMedPubMedCentralCrossRefGoogle Scholar