RESUMEN
Sanger's reaction, which was originally developed for amino acid detection, was utilized for enzyme immobilization. The newly synthesized polymer support, which was called polymer NO2-4-fluorostyrene-divinylbenzene (pNFD), was embedded with a Sanger's reagent-like functional group for immobilizing enzymes covalently under mild reaction conditions. Using Burkholderia cepacia lipase (BCL) as the target enzyme, the immobilization efficiency and activity of pNFD-BCL reached as high as 1.2 mg·g-1 and 33.21 U·g-1 (a specific activity of 27 675 U·g-1), respectively, realizing 90% activity recovery. It also improved the optimal reaction temperature of BCL from 40 to 65 °C, under which its full activity could be retained for 4 h. The new carrier also widened the pH-adaptive range of BCL as 6.5-10.0, allowing the lipase to operate normally in weak acid environment. Reusability of pNFD-BCL was significantly improved as almost no activity and/or enantioselectivity loss was observed in 200 h of triglyceride hydrolysis reaction and 17 batches of ( R, S)-1-phenylethanol resolution reaction.