Leirião P R S, Fonseca L J P, Taipa M A, Cabral J M S, Mateus M
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1049-001 Lisboa, Portugal.
Appl Biochem Biotechnol. 2003 Jul;110(1):1-10. doi: 10.1385/abab:110:1:1.
A hydrophilic polyacrylonitrile (PAN) flat sheet membrane was aminated (8.5 micromol of NH2/mg of dry support) for covalent binding of horseradish peroxidase (HRP), mediated by the soluble carbodiimide l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDC). Silica microbeads derivatized by silanization, to yield an aminated support, and commercial aminated glass microbeads were also coupled to HRP with EDC or activated with glutaraldehyde. The immobilized enzyme activities were determined in a batch enzyme reactor with an external loop, the highest specific immobilized HRP activity being obtained on the glass support (55.8 U/mg of protein). Continuous operational stability studies showed that hydrophilic PAN membrane led to the highest retention of HRP activity after an overall period of 35 h, with a normalized productivity of 59.5 micromol of H2O2 reduced/(h x Uimmob HRP).
通过可溶性碳二亚胺1-乙基-3-(3-二甲基氨基丙基)碳二亚胺（EDC）介导，将亲水性聚丙烯腈（PAN）平板膜胺化（8.5微摩尔NH2/毫克干载体），用于辣根过氧化物酶（HRP）的共价结合。通过硅烷化衍生的二氧化硅微珠产生胺化载体，商业胺化玻璃微珠也用EDC与HRP偶联或用戊二醛活化。在带有外循环的间歇式酶反应器中测定固定化酶活性，在玻璃载体上获得最高的特异性固定化HRP活性（55.8 U/毫克蛋白质）。连续操作稳定性研究表明，亲水性PAN膜在35小时的总时间段后导致HRP活性的最高保留率，归一化生产率为59.5微摩尔H2O2还原/(小时×U固定化HRP)。
