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<p>2-Ketogluconic acid is a valuable organic acid produced via periplasmic glucose oxidation in <i>Pseudomonas</i> species. This study evaluated the production of 2KGA by <i>Pseudomonas reptilivora</i> B-6bs under flask-level conditions using a 23⁻1 fractional factorial design. The effects of glucose concentration (50 to 250&#xa0;g/L), CaCO₃ supplementation (30 to 50&#xa0;g/L), and agitation speed (150 to 230&#xa0;rpm) were investigated in Erlenmeyer flasks containing glucose-based media with peptone and meat extract, which were incubated at 30&#xa0;°C for 48&#xa0;h. 2-Ketogluconic acid was quantified via a calibration curve using 2-phenylenediamine dihydrochloride. Glucose consumption was measured via the 3,5-dinitrosalicylic acid method. Thin-layer chromatography was employed for qualitative identification via a mobile phase of ethyl acetate, methanol, glacial acetic acid, and water (6:1.5:1.5:1). The highest 2-ketogluconic acid production (18.02&#xa0;g/L) occurred at 50&#xa0;g/L glucose, 50&#xa0;g/L CaCO₃, and 150&#xa0;rpm. Under these conditions, the specific growth rate reached 0.049&#xa0;h⁻1, with a product/substrate yield of 0.339&#xa0;g/g and a biomass/substrate yield of 0.361&#xa0;g/g. These results suggest that elevated glucose concentrations hinder assimilation and subsequent conversion to 2KGA in <i>P. reptilivora</i> B-6bs, which is consistent with carbon catabolite repression under high-stress conditions. Consequently, the data underscore that preserving oxidative activity in this strain requires a balance on the basis of moderate substrate concentrations.</p> Graphical Abstract <p></p>

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Metabolic constraints on 2-ketogluconic acid production by Pseudomonas reptilivora B-6bs cultivated at the flask level

  • 期刊论文

  • 发表日期：

    2026年4月10日

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  • Itan Homero Ruiz-Hernandez,
  • Monserrat Campuzano-Vargas,
  • Alisson Ambar Ruiz-Toledo,
  • Juan Carlos González-Hernández

摘要

2-Ketogluconic acid is a valuable organic acid produced via periplasmic glucose oxidation in Pseudomonas species. This study evaluated the production of 2KGA by Pseudomonas reptilivora B-6bs under flask-level conditions using a 23⁻1 fractional factorial design. The effects of glucose concentration (50 to 250 g/L), CaCO₃ supplementation (30 to 50 g/L), and agitation speed (150 to 230 rpm) were investigated in Erlenmeyer flasks containing glucose-based media with peptone and meat extract, which were incubated at 30 °C for 48 h. 2-Ketogluconic acid was quantified via a calibration curve using 2-phenylenediamine dihydrochloride. Glucose consumption was measured via the 3,5-dinitrosalicylic acid method. Thin-layer chromatography was employed for qualitative identification via a mobile phase of ethyl acetate, methanol, glacial acetic acid, and water (6:1.5:1.5:1). The highest 2-ketogluconic acid production (18.02 g/L) occurred at 50 g/L glucose, 50 g/L CaCO₃, and 150 rpm. Under these conditions, the specific growth rate reached 0.049 h⁻1, with a product/substrate yield of 0.339 g/g and a biomass/substrate yield of 0.361 g/g. These results suggest that elevated glucose concentrations hinder assimilation and subsequent conversion to 2KGA in P. reptilivora B-6bs, which is consistent with carbon catabolite repression under high-stress conditions. Consequently, the data underscore that preserving oxidative activity in this strain requires a balance on the basis of moderate substrate concentrations.

Graphical Abstract