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不同碳源对利用代谢工程改造的大肠杆菌生产琥珀酸的影响。

Effect of different carbon sources on the production of succinic acid using metabolically engineered Escherichia coli.

作者信息

Andersson Christian, Hodge David, Berglund Kris A, Rova Ulrika

机构信息

Division of Biochemical and Chemical Process Engineering, Luleå University of Technology, SE-971 87 Luleå, Sweden.

出版信息

Biotechnol Prog. 2007 Mar-Apr;23(2):381-8. doi: 10.1021/bp060301y. Epub 2007 Jan 25.

Abstract

Succinic acid (SA) is an important platform molecule in the synthesis of a number of commodity and specialty chemicals. In the present work, dual-phase batch fermentations with the E. coli strain AFP184 were performed using a medium suited for large-scale industrial production of SA. The ability of the strain to ferment different sugars was investigated. The sugars studied were sucrose, glucose, fructose, xylose, and equal mixtures of glucose and fructose and glucose and xylose at a total initial sugar concentration of 100 g L-1. AFP184 was able to utilize all sugars and sugar combinations except sucrose for biomass generation and succinate production. For sucrose as a substrate no succinic acid was produced and none of the sucrose was metabolized. The succinic acid yield from glucose (0.83 g succinic acid per gram glucose consumed anaerobically) was higher than the yield from fructose (0.66 g g-1). When using xylose as a carbon source, a yield of 0.50 g g-1 was obtained. In the mixed-sugar fermentations no catabolite repression was detected. Mixtures of glucose and xylose resulted in higher yields (0.60 g g-1) than use of xylose alone. Fermenting glucose mixed with fructose gave a lower yield (0.58 g g-1) than fructose used as the sole carbon source. The reason is an increased pyruvate production. The pyruvate concentration decreased later in the fermentation. Final succinic acid concentrations were in the range of 25-40 g L-1. Acetic and pyruvic acid were the only other products detected and accumulated to concentrations of 2.7-6.7 and 0-2.7 g L-1. Production of succinic acid decreased when organic acid concentrations reached approximately 30 g L-1. This study demonstrates that E. coli strain AFP184 is able to produce succinic acid in a low cost medium from a variety of sugars with only small amounts of byproducts formed.

摘要

琥珀酸(SA)是多种商品化学品和特种化学品合成中的重要平台分子。在本研究中,使用适合大规模工业生产SA的培养基对大肠杆菌菌株AFP184进行了双相分批发酵。研究了该菌株发酵不同糖类的能力。所研究的糖类包括蔗糖、葡萄糖、果糖、木糖,以及葡萄糖和果糖、葡萄糖和木糖的等量混合物,总初始糖浓度为100 g L-1。AFP184能够利用除蔗糖外的所有糖类和糖组合来生成生物质和生产琥珀酸盐。以蔗糖为底物时,不产生琥珀酸,且蔗糖未被代谢。葡萄糖的琥珀酸产量(每克厌氧消耗的葡萄糖产生0.83 g琥珀酸)高于果糖(0.66 g g-1)。以木糖为碳源时,产量为0.50 g g-1。在混合糖发酵中未检测到分解代谢物阻遏。葡萄糖和木糖的混合物产生的产量(0.60 g g-1)高于单独使用木糖。发酵葡萄糖与果糖的混合物产生的产量(0.58 g g-1)低于以果糖作为唯一碳源时的产量。原因是丙酮酸产量增加。发酵后期丙酮酸浓度降低。最终琥珀酸浓度在25 - 40 g L-1范围内。检测到的其他产物只有乙酸和丙酮酸,其积累浓度分别为2.7 - 6.7 g L-1和0 - 2.7 g L-1。当有机酸浓度达到约30 g L-1时,琥珀酸产量下降。本研究表明,大肠杆菌菌株AFP184能够在低成本培养基中利用多种糖类生产琥珀酸,且仅形成少量副产物。

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