Thermostabilization of Candida antarctica lipase B by double immobilization: Adsorption on a macroporous polyacrylate carrier and R1 silaffin-mediated biosilicification

摘要: Abstract A large improvement in the thermostability of Candida antarctica lipase B (CALB) was achieved through double immobilization, i.e., physical adsorption and R1 silaffin-mediated biosilicification. The C-terminus CALB fused with silaffin peptide for CALB-R1 fusion protein adsorbed onto a macroporous polyacrylate carrier then subsequently biosilicified tetramethyl orthosilicate (TMOS). After biosilicification, double-immobilized exhibited remarkable thermostability. T 50 60 increased dramatically from 45 to 72 °C that 27, 13.8, 9.8 9.9 °C higher than values free CALB-R1, resin, commercial Novozym 435, 435 treated TMOS, respectively. In addition, time required residual activity be reduced half ( t 1/2 ) immobilized elevated 12.2 385 min, which is over 30 times longer life compared CALB-R1. optimum pH biosilicification determined 5.0, enzyme showed much better reusability physically even after 6 repeated reuses. This R1-mediated approach thermostabilization good basis industrial enzymes are only minimally stabilized by engineering.