Application of β-glucuronidase-immobilised silica gel formulation to microfluidic platform for biotransformation of β-glucuronides
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To improve the efficiency of reactions of β-glucuronidase (GUS)-assisted glucuronic acid (GluA) removal within a microfluidic system.
β-glucuronidase from Helix pomatia was immobilised and characterised in silica-based sol–gel monoliths. Efficiency of the GUS-doped silica monoliths was tested for hydrolysis of p-Nitrophenyl-β-d-glucuronide (pNP–GluA) in both ml-scaled medium via batch reactions and microfluidic environment via continuous-flow reactions. In the microfluidic platform, within a duration of 150 min of continuous operation (flow rate: 1 µL/min), the obtained highest pNP yield was almost 50% higher than that of the corresponding batchwise reaction. However, increased flow rates (3, 5, and 10 µL/min) resulted in lower conversion yields compared to 1 µL/min. The microfluidic platform demonstrated continuous hydrolytic activity for 7 days with considerable reaction yields while using a small amount of the enzyme.
These results revealed that usage of the microreactors has considerable potential to efficiently obtain bioactive GluA-free aglycons from various plant-derived β-glucuronides for pharmaceutical applications.
Keywordsβ-glucuronidase Enzyme immobilisation Glucuronide hydrolysis Microfluidic platform Microenzyme reactor Sol–gel
The financial support provided by the Scientific and Technological Research Council of Turkey (TUBITAK, 113M050) is highly appreciated. Special thanks are offered to Dr Barbaros Cetin from the Bilkent University Microfluidics & Lab-on-a-chip Research Group for microfabrication studies, and Dr Bogdan Parakhonskiy from the Ghent University for his valuable comments on the manuscript.
Supplementary Material Section A—methods of activity measurements for enzyme kinetics and stability.
Supplementary Material Section B—procedures regarding fabrication of the microfluidic apparatus.
Supplementary Fig. 1—Stability of immobilised GUS at 4 and 37 °C.
Compliance with ethical standards
Conflict of interest
The authors have no conflicts of interest in relation to the manuscript.
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