西安交通大学生命科学与技术学院,西安,710049
网络首发:2008-08-10,
纸质出版:2008
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邵卫祥, 莫晓燕, 李黎. 磁性交联核酸酶P1聚集体的制备及性质研究[J]. 西安交通大学学报, 2008,42(8):1035-1039.
邵卫祥, 莫晓燕, 李黎. Preparation and Property of Magnetic Cross-Linked Nuclease P1 Aggregations[J]. 2008, 42(8): 1035-1039.
采用磁性纳米颗粒与酶蛋白共沉淀后经戊二醛交联的方法
制备了磁性交联核酸酶P1聚集体
并且对比分析了游离酶和固定化酶的部分酶学性质.优化的最佳制备条件为:硫酸铵质量浓度为0.8 g/mL
沉淀时间为0.5 h
戊二醛体积分数为0.6%
交联时间为2 h
所制得的固定化酶活性回收率为32.4%.酶学性质研究表明
固定化酶的K
m
值(30.7 mmol/L)明显高于游离酶的(7.27 mmol/L)
二者最适反应温度分别为90 ℃和75 ℃
最适pH值均为5.2
固定化核酸酶P1对热和酸碱的耐受性明显增强
连续反应6次后酶活力仍保留70%
良好的操作稳定性和磁响应性有利于核酸酶P1的工业化应用.
The magnetic cross-linked enzyme aggregations(CLEAs)were prepared by depositing of nuclease P1 with magnetic nanoparticles and cross-linked with glutaraldehyde
and some enzymetic properties were compared between nuclease P1 and CLEAs. The optimal conditions of preparing magnetic CLEAs are sought out:(NH
4
)
2
SO
4
80% saturation
depositing time 0.5 h
glutaraldehyde 0.6%
and cross-linking time 2 h. The activity recovery reaches to 32.4% under the above conditions. Investigation of enzymetic properties suggests that K
m
of magnetic CLEAs(30.7 mmol/L)gets higher than soluble enzyme(7.27 mmol/L)
and the optional reacting temperature of immobilized nuclease P1 is appropriate at 90 ℃
while optional reacting pH approaches to 5.2. Once immobilized
the resistance to heat
acid and alkali of nuclease P1 is remarkably improved
and the magnetic CLEAs maintained 70% activity after six consecutive reactions. Good operational stability and magnetic response are beneficial for industrial utilization of nuclease P1.
武仙山,何立千,叶磊.交联酶聚集体:一种无载体酶固定化方法[J].生物技术,2005, 15(2):90-92.
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任欢鱼,刘蕾,刘勇键.磁流体的制备与性质研究[J].中国粉体技术,2003, 9(1):21-23.
REN Huanyu,LIU Lei,LIU Yongjian. Preparation and properties of magnetic fluid[J].China Powder Science and Technology,2003,9(1):21-23.
莫晓燕,宋威,张芹,等.桔青霉生产核酸酶P1发酵条件优化研究[J].西安交通大学学报,2003, 37(10):1083-1086.
MO Xiaoyan,SONG Wei,ZHANG Qin.Study on the optimization of fermentation conditions for penicillium citrinum producing nuclease P1[J]. Journal of Xi'an Jiaotong University,2003,37(10):1083-1086.
LORENA W, LORENA B, GLORIA F, et al. Cross-linked aggregates of multimeric enzymes:a simple and efficient methodology to stabilize their quaternary structure[J]. Biomacromolecules, 2004(5): 814-817.
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