苯丙氨酸解氨酶仿生固定化及其稳定性研究

崔建东; 梁龙昊; 韩丛; 刘容麟

doi:10.13386/j.issn1002-0306.2015.18.030

苯丙氨酸解氨酶仿生固定化及其稳定性研究

河北省发酵工程技术研究中心生物科学与工程学院河北科技大学

基金项目:

国家自然科学基金(21072041)；河北省自然科学基金(B2014208054)；

详细信息

作者简介:
崔建东(1974-),男,博士,教授,研究方向:生物催化与微生物资源开发,E-mail:cjd007cn@163.com。;

中图分类号: Q814.2
计量
- 文章访问数: 222
- HTML全文浏览量: 20
- PDF下载量: 134
出版历程
- 收稿日期: 2015-01-19

Immobilization of phenylalanine ammonia lyase by biomimetic silica and its stability

Research Center for Fermentation Engineering of Hebei,College of Bioscience and Bioengineering,Hebei University of Science and Technology

摘要

摘要: 利用仿生硅化技术包埋固定化苯丙氨酸解氨酶(PAL),研究固定化条件对PAL固定化的影响及固定化PAL的稳定性。优化的固定化酶制备条件:0.06 m L浓度为6 mg/m L诱导剂聚乙烯亚胺(PEI),25 mmol/L磷酸盐(p H7.0)作为固定化反应介质体系,2 m L浓度为1 mol/L正硅酸甲酯(TMOS)水解液和1 m L(2 U/m L)酶液添加量,所得固定化酶的酶活最大回收率是70%。与游离酶相比,固定化PAL的温度稳定性、p H和储存稳定性,以及变性剂耐受性都有较大提高,重复使用5次,固定化PAL仍能保持初始酶活的40%。
- 苯丙氨酸解氨酶 /
- 仿生硅化 /
- 氧化硅 /
- 固定化酶 /
- 聚乙烯亚胺
Abstract: In this study,phenylalanine ammonia lyase(PAL) was encapsulated by in polyallylamine-mediated biomimetic silica. The conditions for the preparation of encapsulated PAL were optimized. Moreover,stability of the encapsulated PAL was examined. The optimal activity recovery(70%) of PAL were achieved when 0.06 m L polyallylamine(PEI) of 6 mg/m L,2 m L tetramethoxysilane(TMOS) of 1 mol/L and 1 m L PAL(2 U/m L) in 25 mmol/L phosphate buffer(p H7.0) were used. Compared with free PAL,the encapsulated PAL showed better properties in p H,thermal and storage stabilities,as well as the tolerance abilities against denaturants. In additional,the encapsulated PAL still retained 40% of its initial activity after consecutive 5 cycles.
- phenylalanine ammonia lyase /
- biomimetic silica /
- monox /
- Immobilization enzyme /
- polyethyleneimine