Efficient Biosynthesis of 2-keto-D-gluconic Acid by Fed-batch Culture of Metabolically Engineered
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2-keto-d-gluconic acid (2-KGA) is a key precursor for synthesising vitamin C and isovitamin C. However, phage contamination is as constant problem in industrial production of 2-KGA using . holds promise for producing 2-KGA due to impressive resistance to hypertonicity and acids, and high utilisation of glucose. In this study, the 2-KGA synthesis pathway was regulated to enhance production of 2-KGA and reduce accumulation of the by-products 5-keto-d-gluconic acid (5-KGA) and d-gluconic acid (D-GA) in the 2-KGA producer CGMCC 1.49. Knocking out the gene from a competitive pathway and overexpressing the gene from the 2-KGA synthesis pathway via homologous recombination increased the titre of 2-KGA by 63.81% in shake flasks. Additionally, accumulation of 5-KGA was decreased by 63.52% with the resulting -Δ- strain. Using an intermittent fed-batch mode in a 3 L fermenter, 2-KGA reached 235.3 g L with a 91.1% glucose conversion rate. Scaling up in a 15 L fermenter led to stable 2-KGA titre with productivity of 2.99 g L h, 11.99% higher than in the 3 L fermenter, and D-GA and 5-KGA by-products were completely converted to 2-KGA.
Nakashima S, Matsutani M, Kataoka N, Adachi O, Yamashita R, Matsushita K Appl Environ Microbiol. 2025; 91(2):e0250124.
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Tian J, Wei S, Liang W, Wang G Synth Syst Biotechnol. 2024; 10(1):68-75.
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Zeng W, Shan X, Liu L, Zhou J Bioresour Bioprocess. 2024; 9(1):121.
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