LI Zhigang, ZHANG Chaohui, TAN Hai, et al. Enhancing cAMP Fermentation Formation via Glutathione and Auxiliary Energy Substance Coupling Addition[J]. Science and Technology of Food Industry, 2021, 42(20): 119−125. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021010246.
Citation: LI Zhigang, ZHANG Chaohui, TAN Hai, et al. Enhancing cAMP Fermentation Formation via Glutathione and Auxiliary Energy Substance Coupling Addition[J]. Science and Technology of Food Industry, 2021, 42(20): 119−125. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021010246.

Enhancing cAMP Fermentation Formation via Glutathione and Auxiliary Energy Substance Coupling Addition

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  • Received Date: January 31, 2021
  • Available Online: August 10, 2021
  • Objective: To elucidate the cause for ceased cAMP formation during the later stage of fermentation with auxiliary energy substance added and develop coping strategy to further improve cAMP production. Methods: cAMP fermentations with sodium citrate and sodium pyruvate respective addition were conducted in a 7 L fermenter and corresponding fermentation kinetics, cells viability, energy metabolism and ROS levels were analyzed. Finally, a fermentation process with glutathione and auxiliary energy substance coupling added was proposed and conducted for improving cAMP contents. Results: The cAMP formation and cells growth rates in energy substance added conditions were higher than those of control obviously at the initial stage and dropped down at 30 h rapidly to low levels lower than control. The cells viability, ATP/AMP and ETC activities also dropped down at 36 h rapidly to low levels with larger decline than control. The analysis results indicated that reactive oxygen species (ROS) and malonaldehyde contents increased rapidly after 36 h due to the addition of auxiliary energy substance, which were higher than those of control obviously (P<0.01), whereas NADPH/NADP+ dropped sharply at the same time holding at lower levels than control. Finally, with the proposed fermentation process coupling glutathione and auxiliary energy substance added, cell and cAMP concentrations were improved obviously due to higher cell viability. cAMP concentrations in citrate coupling glutathione added batch and pyruvate coupling glutathione added batch achieved 4.05 and 4.32 g/L, respectively, with increments of 15.2% and 22.7% when compared with those of only GSH added batch, and with increments of 21.9% and 30.1% when compared with control(without auxiliary energy substance and glutathione addition). Conclusion: Auxiliary energy substance aroused plentiful ROS formation due to the intensive electron leakage from electron transport chain which caused cell injury and ceased cAMP synthesis. The fermentation process with glutathione and auxiliary energy substance coupling added relieved cell damage from ROS and further enhanced cell viability and cAMP fermentation production.
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  • [1]
    Argyrousi E K, Heckman P R, Prickaerts J, et al. Role of cyclic nucleotides and their downstream signaling cascades in memory function: Being at the right time at the right spot[J]. Neuroscience and Biobehavioral Reviews,2020,113:12−38. doi: 10.1016/j.neubiorev.2020.02.004
    [2]
    Wang D C, Qi J Z, Han W, et al. Kanamycin-induced production of 2’,3’-cyclic AMP in Escherichia coli[J]. Biochemical and Biophysical Research Communications,2020,527(4):854−860. doi: 10.1016/j.bbrc.2020.04.144
    [3]
    Dong J, Fu X M, Wang P F, et al. Construction of industrial baker's yeast with high level of cAMP[J]. Journal of Food Biochemistry,2019,43(7):1−7.
    [4]
    Chen Y, Li S Y, Xiong J, et al. The mechanisms of citrate on regulating the distribution of carbon flux in the biosynthesis of uridine 5’-monophosphate by Saccharomyces cerevisiae[J]. Applied Microbiology and Biotechnology,2010,86:75−81. doi: 10.1007/s00253-009-2287-y
    [5]
    Zhang H, Zhu J Z, Zhu X C, et al. High-level exogenous glutamic acid-independent production of poly-(γ-glutamic acid) with organic acid addition in a new isolated Bacillus subtilis C10[J]. Bioresource Technology,2012,116:241−246. doi: 10.1016/j.biortech.2011.11.085
    [6]
    李志刚, 陈宝峰, 张中华, 等. 辅助能量物质强化环磷酸腺苷发酵合成机制[J]. 中国生物工程杂志,2020,40(1/2):102−108. [Li Z G, Chen B F, ZHANG Z H, et al. The physiological mechanism for enhanced cyclic adenosine monophosphate biosynthesis by auxiliary energy substance[J]. China Biotechnology,2020,40(1/2):102−108.
    [7]
    Chen S X, Chu J, Zhuang Y P, et al. Enhancement of inosine production by Bacillus subtilis through suppression of carbon overflow by sodium citrate[J]. Biotechnology Letters,2005,27:689−692. doi: 10.1007/s10529-005-4686-1
    [8]
    Wang Y L, Wang D H, Wei G Y, et al. Improved co-production of S-adenosylmethionine and glutathione using citrate as an auxiliary energy substrate[J]. Bioresource Technology,2013,131:28−32. doi: 10.1016/j.biortech.2012.10.168
    [9]
    Chen X C, Song H, Fang T, et al. Enhanced cyclic adenosine monophosphate production by Arthrobacter A302 through rational redistribution of metabolic flux[J]. Bioresource Technology,2010,101:3159−3163. doi: 10.1016/j.biortech.2009.12.081
    [10]
    Xia J, Xu Z X, Xu H, et al. The regulatory effect of citric acid on the co-production of poly(ε-lysine) and poly(L-diaminopropionic acid) in Streptomyces albulus PD-1[J]. Bioprocess and Biosystems Engineering,2014,37:2095−2103. doi: 10.1007/s00449-014-1187-4
    [11]
    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[J]. 食品与发酵工业,2018,44(11):154−158. [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[J]. Food and Fermentation Industries,2018,44(11):154−158.
    [12]
    González-Siso M I, Garcia-Leiro A, Tarrio N, et al. Sugar metabolism, redox balance and oxidative stress response in the respiratory yeast Kluyveromyces lactis[J]. Microbial Cell Factories,2009,8:46−49. doi: 10.1186/1475-2859-8-46
    [13]
    Yan P, Sun H B, Lu P Q, et al. Enhancement of ε-poly-L-lysine synthesis in Streptomyces by exogenous glutathione[J]. Bioprocess and Biosystems Engineering,2018,41:129−134. doi: 10.1007/s00449-017-1849-0
    [14]
    Li S, Ji J Y, Hu S J, et al. Enhancement of ε-poly-L-lysine production in Streptomyces griseofuscus by addition of exogenous astaxanthin[J]. Bioprocess and Biosystems Engineering,2020,43:1813−1821. doi: 10.1007/s00449-020-02372-y
    [15]
    李志刚, 顾阳, 陈宝峰, 等. 硝酸盐促进环磷酸腺苷发酵合成的生理机制研究[J]. 食品工业科技,2021,42(1):115−120. [Li Z G, Gu Y, Chen B F, et al. Physiological mechanism for enhanced cyclic adenosine monophosphate biosynthesis by sodium nitrate in Arthrobacter sp. CCTCC M2013431[J]. Science and Technology of Food Industry,2021,42(1):115−120.
    [16]
    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[J]. Scientific Reports,2018,8(1):1246−1249. doi: 10.1038/s41598-017-18889-4
    [17]
    Hatzinger P, Palmer P, Smith R, et al. Applicability of tetrazolium salts for the measurement of respiratory activity and viability of groundwater bacteria[J]. Journal of Microbiological Methods,2003,52(1):47−58. doi: 10.1016/S0167-7012(02)00132-X
    [18]
    Huang J C, Zhong Y J, Liu J, et al. Metabolic engineering of tomato for high-yield production of astaxanthin[J]. Metabolic Engineering,2013,17:59−67. doi: 10.1016/j.ymben.2013.02.005
    [19]
    Liu C B, Pan F, Li Y, et al. A combined approach of generalized additive model and bootstrap with small sample sets for fault diagnosis in fermentation process of glutamate[J]. Microbial Cell Factories,2016,15:1−6. doi: 10.1186/s12934-015-0402-6
    [20]
    徐若烊, 王大慧, 许宏庆, 等. 丙酮酸钠促进S-腺苷蛋氨酸和谷胱甘肽联合高产及其生理机制[J]. 食品工业科技,2018,39(2):113−118. [Xu R Y, Wang D H, Xu H Q, et al. Sodium pyruvate improves the co-production of S-adenosylmethionine and glutathione and its physiological mechanism[J]. Science and Technology of Food Industry,2018,39(2):113−118.
    [21]
    Hara K, Kondo A. ATP regulation in bioproduction[J]. Microbial Cell Factories,2015,14(1):198−207. doi: 10.1186/s12934-015-0390-6
    [22]
    Chen H L, Cao X T, Zhu N Q, et al. A stepwise control strategy for glutathione synthesis in Saccharomyces cerevisiae based on oxidative stress and energy metabolism[J]. World Journal of Microbiology and Biotechnology,2020,36(8):1−8.
    [23]
    Shao H, Tu Y, Wang Y J, et al. Oxidative stress response of Aspergillus oryzae induced by hydrogen peroxide and menadione sodium bisulfite[J]. Microorganisms,2019,7:225−237. doi: 10.3390/microorganisms7080225
    [24]
    Angelova M B, Pashova S B, Spasova B K, et al. Oxidative stress response of filamentous fungi induced by hydrogen peroxide and paraquat[J]. Mycological Research,2005,109(2):150−158. doi: 10.1017/S0953756204001352
    [25]
    Satowa D, Fujiwara R, Uchio S, et al. Metabolic engineering of E. coli for improving mevalonate production to promote NADPH regeneration and enhance acetyl-CoA supply[J]. Biotechnology and Bioengineering,2020,117(7):1−8.
    [26]
    许曼, 江贤章, 黄建忠, 等. 强化类球红细菌辅因子NADPH再生以提高法尼醇的产量[J]. 生物工程学报,2020,36(1):90−99. [Xu M, Jiang X Z, Huang J Z, et al. Reinforcement of Rhodobacter sphaeroides cofactor NADPH to increase the production of farnesol[J]. Chinese Journal of Biotechnology,2020,36(1):90−99.
    [27]
    Zeng X, Chen X S, Gao Y, et al. Continuously high reactive oxygen species generation decreased the specific ε-poly-L-lysine formation rate in fed-batch fermentation using glucose and glycerol as a mixed carbon source[J]. Process Biochemistry,2015,50:1993−2003. doi: 10.1016/j.procbio.2015.09.012
    [28]
    Li Y, Hugenholtz J, Abee T, et al. Glutathione protects Lactococcus lactis against oxidative stress[J]. Applied and Environmental Microbiology,2003,69:5739−5745. doi: 10.1128/AEM.69.10.5739-5745.2003
    [29]
    Penninckx M J. An overview on glutathione in Saccharomyces versus non-conventional yeasts[J]. FEMS Yeast Research,2002,2:295−305.

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