State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China; The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
J Biotechnol. 2014 Jan;169:15-22. doi: 10.1016/j.jbiotec.2013.10.027. Epub 2013 Oct 27.
Commercial production of 2-keto-l-gulonic acid (2-KLG), the immediate precursor of l-ascorbic acid, is by Ketogulonicigenium vulgare in co-culture with Bacillus megaterium. We used flux balance analysis (FBA) to study a genome-scale metabolic model (GSMM) of K. vulgare, iWZ663, and found that K. vulgare is deficient in nutrient biosynthetic pathways. Individually omitting l-glycine, l-cysteine, l-methionine, l-tryptophan, adenine, thymine, thiamine and pantothenate from complete chemically defined medium (CDM), caused biomass formation of K. vulgare to decrease to 1%, 21%, 16%, 1%, 26%, 57%, 73% and 24%, respectively. Based on these results and FBA, a minimal chemically defined medium (MCDM) was developed that supported monoculture of K. vulgare (0.28OD600) and 2-KLG production (3.59g/L), which were similar to those in complete CDM or corn steep liquor powder (CSLP) medium. This study demonstrated the potential of using GSMM and FBA to characterize nutrient requirements, optimize CDM, and study interactions in co-culture.
2-酮基-L-古龙酸（2-KLG）是 L-抗坏血酸的直接前体，其商业生产是通过酮古龙酸菌（Ketogulonicigenium vulgare）与巨大芽孢杆菌（Bacillus megaterium）共培养来实现的。我们使用通量平衡分析（FBA）研究了酮古龙酸菌 iWZ663 的基因组规模代谢模型（GSMM），发现该菌缺乏营养生物合成途径。单独从完全化学定义培养基（CDM）中去除 l-甘氨酸、l-半胱氨酸、l-蛋氨酸、l-色氨酸、腺嘌呤、胸腺嘧啶、硫胺素和泛酸，会导致酮古龙酸菌的生物量形成分别减少到 1%、21%、16%、1%、26%、57%、73%和 24%。基于这些结果和 FBA，开发了一种最小化学定义培养基（MCDM），该培养基支持酮古龙酸菌的单培养（0.28OD600）和 2-KLG 生产（3.59g/L），与完全 CDM 或玉米浆干粉（CSLP）培养基中的培养结果相似。这项研究表明，使用 GSMM 和 FBA 来表征营养需求、优化 CDM 以及研究共培养中的相互作用具有潜力。
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