School of Biology and Chemical Engineering, Jiaxing University, Jiaxing 314001, China.
Appl Biochem Biotechnol. 2010 Mar;160(3):822-30. doi: 10.1007/s12010-009-8608-x. Epub 2009 Apr 21.
In this study, we introduced a new strategy, feeding D-glucose, to overproduce extracellular 5-aminolevulinic acid (ALA) in the recombinant Escherichia coli. We investigated that the D-glucose concentration is dependent on extracellular ALA production. The results indicated that increasing D-glucose concentration in bacteria culture enhanced final cell density and ALA yield and simultaneously decreased the activities of ALA synthase (ALAS) and ALA dehydratase (ALAD); then, the inhibitory effect of D-glucose on ALAS activity was relieved with the metabolism of D-glucose. when 4.0 g/L D-glucose was added at late exponential phase; 1.46 g/L ALA was achieved in shaking culture, which is 47% or 109% higher than the ALA yields with 30 mM levulinic acid of ALAD inhibitor or no inhibitor. In jar fermenter, final extracellular ALA concentration reached 3.1 g/L by feeding with D-glucose.
在这项研究中,我们引入了一种新策略,即喂养 D-葡萄糖,以在重组大肠杆菌中过量生产细胞外 5-氨基乙酰丙酸(ALA)。我们研究了 D-葡萄糖浓度对细胞外 ALA 生产的依赖性。结果表明,增加细菌培养物中的 D-葡萄糖浓度可以提高最终细胞密度和 ALA 产量,同时降低 ALA 合酶(ALAS)和 ALA 脱水酶(ALAD)的活性;然后,随着 D-葡萄糖的代谢,D-葡萄糖对 ALAS 活性的抑制作用得到缓解。当在指数生长期晚期添加 4.0 g/L 的 D-葡萄糖时;在摇瓶培养中可获得 1.46 g/L 的 ALA,比添加 30 mM 乙酰丙酸作为 ALAD 抑制剂或不添加抑制剂时的 ALA 产量分别提高了 47%或 109%。在罐式发酵罐中,通过添加 D-葡萄糖,最终细胞外 ALA 浓度达到 3.1 g/L。