Covalent Immobilization of Lipase AY on Poly (γ-Glutamic acid)

• Main Authors: Shuo-Fen Chang, 張碩芬
• Other Authors: Chwen-Jen Shieh
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/93727807265400346508

碩士 === 大葉大學 === 生物產業科技學系 === 94 === Industrial application of lipase requires efficient methods to immobilize theenzyme for yielding a biocatalyst with greater than free lipase. Lipase AY from Candida rugosa was immobilized on Celite by adsorption and poly γ-glutamic acid (γPGA) by covalent binding, respectively. Response surface methodology (RSM) and 3-level-3-factor fractional factorial design were adopted to evaluate the effects of immobilization variables, such as immobilized time, temperature and enzyme/support ratio on specific activity of immobilization lipase. For lipase AY was immobilized on Celite by adsorption. The optimum immobilization conditions were: immobilization time 59.1 min, immobilization temperature 10.7 °C, enzyme/support ratio 0.5 (w/w) and the highest specific activity was 18.2 U/mg-protein with activity yield 34.1%. In the study of covalent binding, lipase AY was immobilized on γPGA using EDC as a activator. The optimum immobilization conditions were: immobilized time near to 2 h, immobilized temperature 0 °C, enzyme/support ratio about 0.1 (w/w) and the highest specific activity was 96.4 U/mg-protein with activity yield 180.9%. The results showed that the lipase-γPGA has better activity than lipase AY-Celite and free lipase AY and demonstrated the possible industrial application of lipase AY-γPGA for yielding higher production.