Abstract
A dodecane/thermosensitive polymer/water three-liquid-phase system was introduced for enzymatic hydrolysis of penicillin G (Pen G) for 6-aminopenicillanic acid (6-APA). The enzyme was covalently attached to the terminal of PEO–PPO–PEO polymer (L63), which would be transferred into a polymer coacervate phase at high temperature above its “cloud point”. 6-APA was primarily resided in the aqueous phase due to its zwitterionic nature. More than 70% phenylacetic acid (PAA) was transferred into the organic phase using trioctylmethylammonium hydroxide and trihexyl-(tetradecyl)phosphonium bis 2,4,4-trimethylpentylphosphinate ionic liquids (Cyphos IL-104) mixture at pH 5.5, while most of Pen G resided in water. As a result, high operational pH was permitted in three-liquid-phase system, which leads to higher enzymatic activity (120 IU at 40°C) and stability (enzymatic half-time up to 55 h at 60°C) in comparison with the value in butyl acetate/water two-phase system. On the other hand, two products in three-liquid-phase system might be automatically separated from the enzyme sphere into different phases at the same time, which facilitated the reaction equilibrium towards the product’s side with 6-APA productivity of 80% at 42°C, pH 5.5.
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Acknowledgment
This work was financially supported by Innovation Research Group (No.20221603), National Nature Science Foundation of China (No.20490200) and the Fundamental Foundation of China (No.90610007).
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Jiang, Y., Xia, H., Guo, C. et al. Enzymatic Hydrolysis of Penicillin for 6-APA Production in Three-Liquid-Phase System. Appl Biochem Biotechnol 144, 145–159 (2008). https://doi.org/10.1007/s12010-007-8018-x
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DOI: https://doi.org/10.1007/s12010-007-8018-x