Tailoring a robust and recyclable nanobiocatalyst by immobilization of Pseudomonas fluorescens lipase on carbon nanofiber and its application in synthesis of enantiopure carboetomidate analogue

Abstract Pseudomonas fluorescens lipase (PFL) was covalently immobilized on carbon nanofiber (CNF) using 1‑ethyl‑3‑[3‑dimethylaminopropyl] carbodiimide (EDC)/ N ‑hydroxysuccinimide (NHS). Surface functionalization of carbon nanofiber augments dispersibility as well as efficiency of covalent immobilization. Crucial parameters for immobilization such as pH, enzyme-support ratio, reaction time and mixing rate were optimized using one factor at a time (OFAT) approach. The nanobiocatalyst prepared under optimized conditions demonstrated a ten-fold increase in enzyme activity and the advantage of high thermal stability (up to 85 °C) along with 10 cycles of reusability. Subsequently practical application of the nanobiocatalyst was explored in the kinetic resolution of racemic 1‑phenylethanol into ( S )‑1‑phenylethanol [C = 49.1%, ee p  = 99.5%, ee s  = 98.5% and E value = 151.4] followed by Mitsunobu reaction with a substituted pyrrole, giving an enantiopure ( R )-carboetomidate analogue (yield = 83%).