Transcriptomic analysis of Arthrobacter sp. CGMCC 3584 responding to pde gene knockout
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ABSTRACT: Arthrobacter sp. CGMCC 3584 are able to produce high yields of extracellular cyclic adenosine monophosphate (cAMP), which plays a vital role in the field of treatment of disease and animal food, during aerobic fermentation. Comparative transcriptomic analysis revealed that arpde inactivation had two major effects on metabolism: inhibition of glycolysis, PP pathway, and amino acid metabolism; promotion of the purine metabolism and carbon flux from the precursor PRPP, which benefited cAMP production.
Project description:Arthrobacter sp. CGMCC 3584 are able to produce high yields of extracellular cyclic adenosine monophosphate (cAMP), which plays a vital role in the field of treatment of disease and animal food, during aerobic fermentation. DNA array-based transcriptional analysis of Arthrobacter cells was conducted to elucidate the higher productivity of cAMP under high oxygen supply. Results showed that 14.1% and 19.3% of the whole genome genes were up-regulated and down-regulated notably, respectively. The largest group with altered transcriptional levels belonged to the group involved in carbohydrate transport and metabolism. Other large functional groups of differentially expressed genes changed significantly included amino acid transport and metabolism, inorganic ion transport and metabolism and transcription.
Project description:Arthrobacter sp. CGMCC 3584 is able to produce high yields of extracellular cyclic adenosine monophosphate (cAMP), which plays a vital role in the field of treatment of disease and animal food, during aerobic fermentation. However, the molecular basis of cAMP production in Arthrobacter species is rarely explored. Here, for the first time, we report the comparative transcriptomic and proteomic study of Arthrobacter cells to elucidate the higher productivity of cAMP under high oxygen supply. We finally obtained 14.1% and 19.3% of the Arthrobacter genome genes which were up-regulated and down-regulated notably, respectively, with high oxygen supply, and identified 54 differently expressed proteins. Our results revealed that high oxygen supply had two major effects on metabolism: inhibition of glycolysis, pyruvate metabolism, nitrogen metabolism, and amino acid metabolism (histidine, branched-chain amino acids and glutamate metabolism); enhancement of the tricarboxylic acid cycle and purine metabolism. We also found that regulation of adenylate cyclase and phosphodiesterase was not significant under high oxygen supply, suggesting efficient cAMP export might be important in cAMP production. These findings may contribute to further understanding of capacities of Arthrobacter species and would be highly useful in genetic regulation for desirable production.
Project description:Purpose: The goal of this study is compare the effect of phbC gene in curdlan synthesis in Agrobacterium sp. CGMCC 11546. methods: The transcriptional and metabolomics analysis the function of phbC in Agrobacterium sp. CGMCC 11546. Results:The transcriptional and metabolomics showed that the decrease of curdlan production in the ΔphbC mutants may be caused by the insufficient supply of energy ATP conclusion:phbC play an important role in curdlan synthesis in Agrobacterium sp. CGMCC 11546
Project description:Purpose: The goal of this study is compare the effect of glnA gene in curdlan synthesis in Agrobacterium sp. CGMCC 11546. methods: The transcriptional and metabolomics analysis the function of glnA in Agrobacterium sp. CGMCC 11546. Results: The transcriptional and metabolomics showed that the decrease of curdlan production in the ΔglnA mutants may be caused by the insufficient supply of energy ATP conclusion: glnA play an important role in curdlan synthesis in Agrobacterium sp. CGMCC 11546
Project description:Purpose: The goal of this study is compare the effect of MetH and MetZ gene in curdlan synthesis in Agrobacterium sp. CGMCC 11546. methods: The transcriptional and metabolomics analysis the function of metH and metZ in Agrobacterium sp. CGMCC 11546. Results: The transcriptional and metabolomics showed that the decrease of curdlan production in the ΔmetH and ΔmetZ mutants may be caused by the insufficient supply of energy ATP conclusion: MetH and MetZ play an important role in curdlan synthesis in Agrobacterium sp. CGMCC 11546
Project description:We conducted transcriptome analysis of Komagataeibacter europaeus CGMCC 20445 samples under different acidity conditions to elucidate the changes in differentially expressed genes throughout the fermentation process.
Project description:Proteomic Analysis. The proteomic expression of CGMCC 6315 under different nutrient concentration conditions was investigated by isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics. The CGMCC 6315 was cultured in LB broth and 1/15LB broth as described above, after which the strains were collected by centrifugation at 10,000× g for 10 min at 4°C. Protein extraction, digestion, iTRAQ labeling and peptide fractionation were performed using the protocol described by Jin et al. 29. Protein identification was conducted using a LC-20AD nano-HPLC instrument (Shimadzu, Kyoto, Japan) equipped with a Q EXACTIVE tandem mass spectrometer (Thermo Fisher Scientific, San Jose, CA, USA) for data-dependent acquisition detection by nano-electrospray ionization. The raw MS/MS data were converted into MGF format by the thermo scientific tool Proteome Discoverer, and the exported MGF files were searched using Mascot (version 2.3.02) against the selected database containing 7546 CGMCC 6315 coding genes. The IQuant software was used for quantitative analysis of the labeled peptides with isobaric tags. Fold changes of >1.7 with p-values <0.05 were used as a cut off for differentially regulated proteins.
Project description:We report the analysis of differentially gene expression after 7 hours and 24 hours fermentation of curdlan in Agrobacterium sp. CGMCC 11546.