The present work reports covalent immobilization of Pseudomonas fluorescens lipase (PFL) on functionalized multiwalled carbon nanotubes (MWCNTs) as a nanobiocatalyst (NBC). This nanobiocatalyst facilitates efficient kinetic resolution of (RS)-1-phenylethanol into (S)-1-phenylethanol [C = 49.7%, ee(p) = 99.5%, ee(s) = 98.1% and E value = 191.4]. The immobilized preparation (MWCNTs-PFL) showed tenfold increase in activity, thermal stability upto 80 degrees C and recyclability (8 cycles). MWCNTs-PFL nanobioconjugate demonstrated better stability and enhanced activity compared to covalently immobilized PFL on other matrices (silver nanoparticles, gold nanoparticles and chitosan beads) used for the study. A statistical design [response surface methodology (RSM)] employed for the optimization of enzyme immobilization parameters made this study statistically more significant. Overall, the newly developed nanobiocatalyst has applications towards the kinetic resolution of racemic compounds. (C) 2017 Elsevier Ltd. All rights reserved.