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中国精品科技期刊2020
李志刚,张朝辉,谭海,等. 辅助能量物质与谷胱甘肽协同作用促进环磷酸腺苷发酵生产[J]. 食品工业科技,2021,42(20):119−125. doi: 10.13386/j.issn1002-0306.2021010246.
引用本文: 李志刚,张朝辉,谭海,等. 辅助能量物质与谷胱甘肽协同作用促进环磷酸腺苷发酵生产[J]. 食品工业科技,2021,42(20):119−125. doi: 10.13386/j.issn1002-0306.2021010246.
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.

辅助能量物质与谷胱甘肽协同作用促进环磷酸腺苷发酵生产

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

  • 摘要: 目的:探究添加辅助能量物质导致环磷酸腺苷(cAMP)发酵后期产物合成停滞的原因,针对性解除限制因素,进一步提高产物产量。方法:在7 L发酵罐上进行添加辅助能量物质的发酵实验,对发酵动力学参数、细胞活性、能量代谢水平以及活性氧(ROS)含量等进行分析,根据分析结果进行了耦合添加谷胱甘肽和辅助能量物质的发酵实验。结果:单独添加柠檬酸钠、丙酮酸钠批次的产物合成和菌体生长速率在发酵前期均明显高于对照,约30 h后快速下降,降至低于对照批次的低水平;菌体活性、ATP/AMP以及电子呼吸链活性在发酵36 h后也剧烈下降,下降幅度远远大于对照批次。添加辅助能量物质导致ROS和丙二醛在发酵36 h后迅速积累,极显著(P<0.01)高于对照批次,而NADPH/NADP+却快速下降,明显低于对照。耦合添加辅助能量物质和谷胱甘肽的发酵批次中,细胞活性得到较大提升,cAMP产量得到进一步提高。柠檬酸钠耦合GSH批次与丙酮酸钠耦合GSH批次的cAMP产量分别达到4.05和4.32 g/L,比单独添加GSH批次分别提高了15.2%和22.7%,与对照(无辅助能量物质和谷胱甘肽添加)相比分别提高了21.9%和30.1%。结论:辅助能量物质强化能量代谢促进产物合成的同时,增加了电子呼吸链中电子泄漏的几率,大量活性氧产生超出细胞承受能力,导致发酵后期细胞生长和产物合成的停滞。辅助能量物质与谷胱甘肽协同作用,有效缓解了氧化胁迫,提高细胞活性,进一步促进产物发酵生产。

     

    Abstract: 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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