Novel Sanger’s Reagent-like Styrene Polymer for the Immobilization of Burkholderia cepacia Lipase

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.