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Knockout of pde gene in Arthrobacter sp. CGMCC 3584 and transcriptomic analysis of its effects on cAMP production

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Abstract

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.

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Acknowledgements

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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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing Tech University, No. 5, Xinmofan Road, Nanjing, 210009, People’s Republic of China

    Huanqing Niu, Chenjie Zhu, Yong Chen, Nan Gao, Hanjie Ying & Dong Liu

  2. National Engineering Technique Research Center for Biotechnology, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30, Puzhu South Road, Nanjing, 211816, People’s Republic of China

    Huanqing Niu, Xinzeng Sun, Jiarui Song, Chenjie Zhu, Yong Chen, Nan Gao, Hanjie Ying & Dong Liu

  3. Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, Wuhan, People’s Republic of China

    Xudong Qu

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  1. Huanqing Niu
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Niu, H., Sun, X., Song, J. et al. Knockout of pde gene in Arthrobacter sp. CGMCC 3584 and transcriptomic analysis of its effects on cAMP production. Bioprocess Biosyst Eng 43, 839–850 (2020). https://doi.org/10.1007/s00449-019-02280-w

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