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利用乙酸作为好氧碳源生产琥珀酸的大肠杆菌 NZN111 的工艺开发。

Process development of succinic acid production by Escherichia coli NZN111 using acetate as an aerobic carbon source.

机构信息

State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.

出版信息

Enzyme Microb Technol. 2011 Oct 10;49(5):459-64. doi: 10.1016/j.enzmictec.2011.07.006. Epub 2011 Jul 30.

Abstract

Escherichia coli strain NZN111 could convert glucose to succinic acid efficiently in anaerobic conditions after the induction of gluconeogenic carbon sources in aerobic conditions. Acetate shows a strong effect on both yield and productivity of succinic acid. In this study, the fed-batch process of succinic acid production by NZN111 using acetate in a chemically defined medium in the aerobic stage was investigated and developed. Increasing cell density could increase succinic acid with a productivity of 3.97 g/(Lh) in the first 8h of the anaerobic phase with an overall yield of 1.42 mol/mol glucose in a 5L fermentor. However, there was strong repression from succinic acid in the later anaerobic stage. When succinic acid exceeded 30 g/L, the glucose consumption rate began to drop sharply along with the succinic acid production rate. Supplementation with glucose from 30 to 70 g/L in the anaerobic stage showed little effect on succinic acid production. Acetic acid and pyruvic acid accumulated had no effect on succinic acid formation because of their low concentration. With acetate as the sole carbon source for aerobic cultivation in the following scale-up, 60.09 g/L of succinic acid was produced with a yield of 1.37 mol/mol in a 50 L bioreactor.

摘要

在有氧条件下诱导生成葡萄糖碳源后,大肠杆菌 NZN111 菌株能够在厌氧条件下高效地将葡萄糖转化为琥珀酸。乙酸对琥珀酸的产率和生产效率都有很强的影响。在这项研究中,我们研究并开发了在有氧阶段使用乙酸的补料分批工艺,在化学定义的培养基中生产琥珀酸。在 5L 发酵罐中,增加细胞密度可以在厌氧阶段的前 8 小时内提高琥珀酸的产量,达到 3.97 g/(L·h),整体产率为 1.42 mol/mol 葡萄糖。然而,在后期的厌氧阶段,琥珀酸受到强烈抑制。当琥珀酸超过 30g/L 时,葡萄糖消耗速率开始随着琥珀酸生产速率的下降而急剧下降。在厌氧阶段补充 30 到 70g/L 的葡萄糖对琥珀酸的生产几乎没有影响。由于浓度低,乙酸和丙酮酸的积累对琥珀酸的形成没有影响。在随后的放大规模中,以乙酸为唯一的有氧培养碳源,在 50L 生物反应器中生产了 60.09g/L 的琥珀酸,摩尔产率为 1.37mol/mol。

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