耐热β-葡萄糖苷酶固定化研究

A Study of Thermostable β-glucosidase Immobilization

本文借助原核表达系统,成功制备了耐热β-葡萄糖苷酶,酶活性可达0.745 k,以经戊二醛交联的DEAE纤维素作为载体固定化耐热β-葡萄糖苷酶,并优化固定化条件,结果显示戊二醛浓度为4%,加酶量为0.5 mL时,固定化效果最好,经测定,固定化酶的酶活回收率可达到84.6%,且载体材料耐高温。本研究实现了经济、安全、高效的耐热β-葡萄糖苷酶制备方法,以及耐热β-葡萄糖苷酶的固定化条件优化,为探索适合商业化生产京尼平的制备技术提供有力的数据支撑。

The recombinant plasmid pHY was transformed into E.coli AS1.1739.The thermostableβ-glucosidase solution is made through a series of steps that is the expansion of LB culture medium,centrifuged,broken cells,heating and so on.The UV spectrophotometry used the ONPG as substrate is used to measure per milliliter of crude enzyme solution absorbance.It is 0.745 k.The protein concentration of the crude enzyme solution measured UV absorption is 1.61 mg/ml.Used DEAE-cellulose cross-linked with glutaraldehyde as a carrier immobilize glycosidases and study the optimization of conditions.The results showed that the recovery rate of the activity of immobilized enzyme can be achieved 84.6%and the carrier material is resistant to high temperature.When the glutaraldehyde concentration was 4%and the enzyme solution is 0.5 mL,the immobilized result is better.However,the operation stability of immobilized enzyme was not so good,need to further optimize the conditions.