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| Global Gene Transcriptome Analysis for Enhanced Cyclic Adenosine Monophosphate Fermentation Performance by Polyphosphates |
LU Nan-xun1,Wang Li-wei1,LIU Mei-xiu1,ZHANG Zhong-hua1,CHANG Jing-ling1,2,LI Zhi-gang1,2,**( ) |
1. School of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang 453003, China
2. Collaborative Innovation Center of Modem Biological Breeding of Henan Province, Xinxiang 453003, China |
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Abstract Objective: To explore the mechanism for enhanced cAMP fermentation production by polyphosphates, Arthrobacter sp. CCTCC 2013431 culture was carried out under low-polyphosphates addition condition as the starting strain. Methods: Fermentations with/without hexametaphosphate addition were conducted in a 7 L bioreactor and the fermentation performance, global gene transcriptome, key enzymes activities together with important metabolites levels were analyzed systematically. Results: With 2 g/L-broth sodium hexametaphosphate added at 24 h, cAMP concentration reached 3.64 g/L with an increment of 33.82% higher than that of control group and the fermentation performance was also promoted obviously. Transcriptome analysis showed that 227 genes were up-regulated significantly and 265 genes were down-regulated significantly due to the addition of hexametaphosphate. For glycometabolism, the transcription levels of key enzyme genes in pentose phosphate pathway and cAMP synthesis pathway were enhanced significantly and for energy metabolism the transcription levels of complex Ⅲ, complex Ⅳ as well as F0F1-ATPase in electron transport chain and polyphosphate kinase gene were also increased significantly by which sufficient carbon skeleton and ATP were provided for cAMP biosynthesis. In addition, transcription levels of reductase genes, such as thioredoxin, catalase and CLP protease, were also increased significantly whereby intracellular redox balance was maintained conducive to cell metabolism and product synthesis. Finally, the activities of pyruvate kinase, 6-phosphoglucose dehydrogenase, adenylosuccinate synthetase, adenylate cyclase, catalase, polyphosphate kinase and intracellular ROS, ATP and NADPH levels under different fermentation conditions were measured to further support the transcriptome analysis results. Conclusion: Sodium hexametaphosphate addition enhanced the carbon flux distribution in pentose phosphate pathway and cAMP synthesis pathway and energy metabolism for ATP synthesis. At the same time, intracellular redox balance was also maintained. Furthermore, cAMP fermentation synthesis and accumulation was promoted significantly.
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Received: 29 October 2022
Published: 04 May 2023
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| [1] |
王瑶函, 吴蕊. cAMP对高血压鼠血管平滑肌细胞增殖的影响. 中国现代医学杂志, 2018, 28(28): 13-20.
|
|
|
| [1] |
Wang Y H, Wu R. Effect of cAMP on proliferation of vascular smooth muscle cells in hypertensive rats. China Journal of Modern Medicine, 2018, 28(28): 13-20.
|
|
|
| [2] |
文川, 马夫天, 万伍卿. cAMP反应元件结合蛋白/Bcl-2在小儿急性白血病骨髓细胞中的表达及意义. 中国当代儿科杂志, 2010, 12(3): 177-180.
|
|
|
| [2] |
Wen C, Ma F T, Wan W Q. Expression of CREB/Bcl-2 in bone marrow mononuclear cells of children with acute leukemia. Chinese Journal of Contemporary Pediatrics, 2010, 12(3): 177-180.
|
|
|
| [3] |
Wartchow K M, Schmid B, Tripal P, et al. Treatment with cyclic AMP activators reduces glioblastoma growth and invasion as assessed by two-photon microscopy. Cells, 2021, 10(3): 556.
doi: 10.3390/cells10030556
|
|
|
| [4] |
Niu H Q, Sun X Z, Song J R, et al. Knockout of pde gene in Arthrobacter sp. CGMCC 3584 and transcriptomic analysis of its effects on cAMP production. Bioprocess and Biosystems Engineering, 2020, 43(5): 839-850.
doi: 10.1007/s00449-019-02280-w
|
|
|
| [5] |
Chen Y W, Liao Y, Kong W Z, et al. ATP dynamic regeneration strategy for enhancing co-production of glutathione and S-adenosylmethionine in Escherichia coli. Biotechnology Letters, 2020, 42(12): 2581-2587.
doi: 10.1007/s10529-020-02989-9
|
|
|
| [6] |
李志刚, 陈宝峰, 方智博, 等. 基于柠檬酸盐与次黄嘌呤偶合添加的环磷酸腺苷发酵工艺. 食品与发酵工业, 2018, 44(11): 154-158.
|
|
|
| [6] |
Li Z G, Chen B F, Fang Z B, et al. A novel fermentation process for cyclic adenosine monophosphate production based on citrate coupling hypoxanthine addition in pulses. Food and Fermentation Industries, 2018, 44(11): 154-158.
|
|
|
| [7] |
Niu H Q, Wang J Z, Zhuang W, et al. Comparative transcriptomic and proteomic analysis of Arthrobacter sp. CGMCC 3584 responding to dissolved oxygen for cAMP production. Scientific Reports, 2018, 8(1): 1-13.
|
|
|
| [8] |
Baumgart M, Unthan S, Kloβ R, et al. Corynebacterium glutamicum chassis C1*: building and testing a novel platform host for synthetic biology and industrial biotechnology. ACS Synthetic Biology, 2018, 7(1): 132-144.
doi: 10.1021/acssynbio.7b00261
pmid: 28803482
|
|
|
| [9] |
Reddy G K, Wendisch V F. Characterization of 3-phosphoglycerate kinase from Corynebacterium glutamicum and its impact on amino acid production. BMC Microbiology, 2014, 14(1): 54-63.
doi: 10.1186/1471-2180-14-54
|
|
|
| [10] |
Yu L J, Wu J R, Liu J, et al. Enhanced curdlan production in Agrobacterium sp. ATCC 31749 by addition of low-polyphosphates. Biotechnology and Bioprocess Engineering, 2011, 16(1): 34-41.
doi: 10.1007/s12257-010-0145-5
|
|
|
| [11] |
Müller W E G, Schröder H C, Wang X H. Inorganic polyphosphates as storage for and generator of metabolic energy in the extracellular matrix. Chemical Reviews, 2019, 119(24): 12337-12374.
doi: 10.1021/acs.chemrev.9b00460
pmid: 31738523
|
|
|
| [12] |
Chen Y W, Cao Y T, Kong W Z, et al. Enhanced glutathione production by bifunctional enzyme coupling with ydaO-based ATP regulating system in Escherichia coli. Journal of Functional Foods, 2020, 75: 104211.
|
|
|
| [13] |
Wang J Z, Zheng C, Zhang T Y, et al. Novel one-pot ATP regeneration system based on three-enzyme cascade for industrial CTP production. Biotechnology Letters, 2017, 39(12): 1875-1881.
doi: 10.1007/s10529-017-2427-x
pmid: 28861634
|
|
|
| [14] |
Chandrashekhar K, Kassem I I, Nislow C, et al. Transcriptome analysis of Campylobacter jejuni polyphosphate kinase (ppk1 and ppk2) mutants. Virulence, 2015, 6(8): 814-818.
doi: 10.1080/21505594.2015.1104449
pmid: 26537695
|
|
|
| [15] |
陈宝峰, 李志刚, 张中华, 等. 低聚磷酸盐与次黄嘌呤偶合添加提高环磷酸腺苷发酵性能. 中国生物工程杂志, 2019, 39(8): 25-31.
|
|
|
| [15] |
Chen B F, Li Z G, Zhang Z H, et al. Enhanced cyclic adenosine monophosphate production by coupling addition of low-polyphosphate and hypoxanthine. China Biotechnology, 2019, 39(8): 25-31.
|
|
|
| [16] |
李志刚, 陈宝峰, 方智博, 等. 基于柠檬酸盐与次黄嘌呤偶合添加的环磷酸腺苷发酵工艺. 食品与发酵工业, 2018, 44(11): 154-158.
|
|
|
| [16] |
Li Z G, Chen B F, Fang Z B, et al. A novel fermentation process for cyclic adenosine monophosphate production based on citrate coupling hypoxanthine addition in pulses. Food and Fermentation Industries, 2018, 44(11): 154-158.
|
|
|
| [17] |
李志刚, 顾阳, 谭海, 等. 氨茶碱与柠檬酸盐协同作用促进环磷酸腺苷发酵生产. 中国生物工程杂志, 2021, 41(7): 50-57.
|
|
|
| [17] |
Li Z G, Gu Y, Tan H, et al. Enhanced cyclic adenosine monophosphate fermentation production by aminophylline and citrate coupling addition. China Biotechnology, 2021, 41(7): 50-57.
|
|
|
| [18] |
谭海. 环磷酸腺苷补救合成的限制因素及高产发酵策略研究. 新乡: 河南科技学院, 2022.
|
|
|
| [18] |
Tan H. Causes analysis for limited cAMP synthesis via salvage pathway and development of high yield fermentation strategy. Xinxiang: Henan Institute of Science and Technology, 2022.
|
|
|
| [19] |
顾阳. 辅助能量物质强化ATP的合成提高cAMP发酵水平的研究. 新乡: 河南科技学院, 2021.
|
|
|
| [19] |
Gu Y. The study of enhancing ATP synthesis with auxiliary energy substance to improve cAMP fermentation level. Xinxiang: Henan Institute of Science and Technology, 2021.
|
|
|
| [20] |
张文静, 丑天胜, 刘芳, 等. 金针菇戊糖磷酸途径的关键基因表达分析. 基因组学与应用生物学, 2019, 38(12): 5542-5549.
|
|
|
| [20] |
Zhang W J, Chou T S, Liu F, et al. Expression analysis of key genes in pentose phosphate pathway of Flammulina velutipes. Genomics and Applied Biology, 2019, 38(12): 5542-5549.
|
|
|
| [21] |
Ishige K, Noguchi T. Inorganic polyphosphate kinase and adenylate kinase participate in the polyphosphate: AMP phosphotransferase activity of Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(26): 14168-14171.
|
|
|
| [22] |
刘洋, 牟庆璇, 石雅南, 等. 微生物细胞工厂的代谢调控. 生物工程学报, 2021, 37(5): 1541-1563.
|
|
|
| [22] |
Liu Y, Mou Q X, Shi Y N, et al. Metabolic regulation in constructing microbial cell factories. Chinese Journal of Biotechnology, 2021, 37(5): 1541-1563.
|
|
|
|
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