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

Niu, Huanqing; Sun, Xinzeng; Song, Jun Young; Zhu, Chenjie; Chen, Yong; Gao, Nan; Qu, Xudong; Ying, Hanjie

doi:10.1007/s00449-019-02280-w

Cited by 5 publications

(2 citation statements)

References 33 publications

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“…In addition to that, only a few Arthrobacter species have been genetically modified and were studied due to the few existing genetic engineering tools. Nevertheless, although transposon mutagenesis systems and homologous recombination systems have already been developed and performed to create mutants of Arthrobacter strains, genetic manipulation is always time consuming and labor intensive [ 12 ]. For example, knocking out a gene via homologous recombination requires typically two separated crossover steps.…”

Section: Introductionmentioning

confidence: 99%

Engineered CRISPR/Cas9 System for Transcriptional Gene Silencing in Arthrobacter Species Indicates Bacterioruberin is Indispensable for Growth at Low Temperatures

Flegler

Lipski

2022

Curr Microbiol

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Pink-pigmented Arthrobacter species produce the rare C50 carotenoid bacterioruberin, which is suspected to be part of the cold adaptation mechanism. In silico analysis of the repertoire of genes encoded by the Arthrobacter agilis and Arthrobacter bussei genome revealed the biosynthetic pathway of bacterioruberin. Although genetic analysis is an essential tool for studying the physiology of Arthrobacter species, genetic manipulation of Arthrobacter is always time and labor intensive due to the lack of genetic engineering tools. Here we report the construction and application of a CRISPR/deadCas9 system (pCasiART) for gene silencing in Arthrobacter species. The engineered system pCasiART is suitable for the Golden Gate assembly of spacers, enabling rapid and accurate construction of adapted systems. In addition, pCasiART has been developed to provide an efficient transcription inhibition system for genome-wide gene silencing. The gene silencing of the phytoene synthase (CrtB), the first enzyme in bacterioruberin biosynthesis, suppressed bacterioruberin biosynthesis in Arthrobacter agilis and Arthrobacter bussei, resulting in a lack of pink pigmentation, reduction of biomass production, and growth rates at low temperatures.

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Section: Introductionmentioning

confidence: 99%

Engineered CRISPR/Cas9 System for Transcriptional Gene Silencing in Arthrobacter Species Indicates Bacterioruberin is Indispensable for Growth at Low Temperatures

Flegler

Lipski

2022

Curr Microbiol

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“…Among these approaches, fermentation production by microorganism has attracted increasing attention, because the process is economical, sustainable and environmental-friendly as compared with complicated chemical synthesis. As far, studies concerning cAMP fermentation mainly focus on strain screening (Song et al 2010), genetic modi cation (Niu et al 2020) and metabolic regulation on the fermentation process (Niu et al 2013(Niu et al , 2018. cAMP is synthesized from ATP directly catalyzed by adenylate cyclase in living cells and abundant ATP supply is bene cial for cAMP production (Li et al 2018).…”

Section: Introductionmentioning

confidence: 99%

Enhanced endogenous amino acids and energy metabolism level for cAMP biosynthesis by Arthrobacter sp. CCTCC 2013431 with citrate as cosubstrate

Chen

et al. 2021

Biotechnol Lett

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Objectives In our previous study, citrate was used as auxiliary energy substance for improving cAMP fermentation performance, however, the regulation mechanism of citrate on improved cAMP contents was not clear. To elucidate the regulation mechanism, cAMP fermentations with/without citrate addition were conducted in a 7 L fermentor using Arthrobacter sp. CCTCC 2013431 and assays on key enzymes activities, energy metabolism level, amino acids contents and peroxidation level were performed.Results With 3 g/L-broth sodium citrate added, cAMP concentration and conversion yield from glucose reached 4.34 g/L and 0.076 g/g which were improved by 30.7% and 29.8%, respectively, when compared with those of control. Citrate changed carbon ux distribution among different routes and more carbon ux was directed into pentose phosphate pathway bene cial to cAMP synthesis. Meanwhile, energy metabolism together with precursor amino acids levels were improved signi cantly owing to strengthened metabolic intensity of tricarboxylate cycle by exogenous citrate utilization which provided energy and substance basis for cAMP production. Moreover, higher glutamate synthesis and oxidative stress caused by citrate addition consumed excessive NADPH derived from pentose phosphate pathway by which feedback suppression for pentose phosphate pathway was relieved e ciently.Conclusion Citrate promoted cAMP fermentation production by Arthrobacter sp. CCTCC2013431 due to enhanced precursor amino acids, energy metabolism level and relieved feedback suppression for pentose phosphate pathway.

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Enhanced ATP and antioxidant levels for cAMP biosynthesis by Arthrobacter sp. CCTCC 2013431 with polyphosphate addition

Tan

et al. 2021

Biotechnol Lett

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

Cited by 5 publications

References 33 publications

Engineered CRISPR/Cas9 System for Transcriptional Gene Silencing in Arthrobacter Species Indicates Bacterioruberin is Indispensable for Growth at Low Temperatures

Engineered CRISPR/Cas9 System for Transcriptional Gene Silencing in Arthrobacter Species Indicates Bacterioruberin is Indispensable for Growth at Low Temperatures

Enhanced endogenous amino acids and energy metabolism level for cAMP biosynthesis by Arthrobacter sp. CCTCC 2013431 with citrate as cosubstrate

Enhanced ATP and antioxidant levels for cAMP biosynthesis by Arthrobacter sp. CCTCC 2013431 with polyphosphate addition

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