CIESC Journal

• 生物化工 • 上一篇    下一篇

Lipozyme TL 100 L的固定化及其催化合成蔗糖-6-乙酯

杨绪娥,郑 璞,孙志浩,倪 晔   

  1. 江南大学生物工程学院工业生物技术教育部重点实验室,江苏 无锡 214122
  • 出版日期:2011-11-05 发布日期:2011-11-21

Immobilization of Lipozyme TL 100 L and catalytic synthesis of sucrose-6-acetate

YANG Xu’e,ZHENG Pu,SUN Zhihao,NI Ye   

  1. The Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,Jiangsu,China
  • Online:2011-11-05 Published:2011-11-21

摘要:

采用树脂吸附法固定脂肪酶Lipozyme TL 100 L,并用于催化合成蔗糖-6-乙酯(S-6-a)。比较7种树脂对Lipozyme TL 100 L的吸附与固定化酶合成S-6-a能力,选择非极性大孔树脂HZ-802为固定化载体。固定化酶在吸附温度30 ,蛋白吸附量50.4758.1 mg/g树脂,相对水含量小于5%时,S-6-a产率达80%以上。用异丙醇以体积比0.51处理Lipozyme TL 100 L,固定化酶的S-6-a产率可达95.4%。固定化酶反复使用8次,S-6-a产率仍有34.2%。与商品固定化酶Lipozyme TLIM相比,HZ-802吸附法制备的固定化酶催化转酯反应的活化能降低了6.47 kJ/mol

Abstract:

Immobilization of Lipozyme TL 100 L on resin was realized by adsorptionand its application in sucrose-6-acetateS-6-asynthesis was then investigated. The results showed that nonpolar macroporous resin of HZ-802 was the best carrier among 7 kinds of resins by comparing both protein adsorptivity and transesterification capacity. The S-6-a yield was attained more than 80% in the condition of immobilization temperature 30 adsorption capacity between 50.4758.1 mg/g resinand immobilized enzyme relative water content under 5%. Disposing Lipozyme TL 100 L with 2-propanol at the volume ratio of 0.51the S-6-a yield of the immobilized enzyme reached 95.4%. And the yield still remained 34.2% when the immobilized enzyme was repeated 8 times. The transesterification followed Ping-Pong Bi Bi mechanism by the studies of apparent michaelis constant Km′ at different substrate concentrations. Activation energy of the immobilized enzyme by adsorption on HZ-802 was reduced 6.47 kJ/mol compared with the commercial Lipozyme TLIM.