Enzymatic Properties of Magnetic Chitosan Microspere-Immobilized Alkaline Protease

Abstract

Abstract: Alkaline protease was immobilized onto magnetic chitosan microspheres prepared from Fe3O4 and chitosan by crosslinking with glutaraldehyde and its appearance and structural properties were analyzed. The results showed that the magnetic chitosan microspheres had spherical appearance with a particle size increasing from 15 to 20 nm after immobilization. FT-IR analysis revealed that Fe3O4 was well wrapped up in the magnetic chitosan microspheres. The magnetic chitosan microspheres had complete crystal structure, good magnetic response and strong superparamagnetic behaviors before and after immobilization of alkaline protease. The optimal reaction temperature and pH for the immobilized enzyme were 50 ℃ and 11 compared with 60 ℃ and 10, respectively, for the free enzyme. The Km was 5.85 ×10-4 mg/mL for the immobilized enzyme and 6.06 ×10-4 mg/mL for its free counterpart.

Key words: magnetism, chitosan, immobilization, alkaline protease, enzymatic property

CLC Number:

ts201

References

[1]林敏刚.碱性蛋白酶在水解植物蛋白中的应用[J ]. 中国油脂, 2009,34 (12), 30～33
[2]杨柳,江连洲, 李杨等.蛋白酶对水酶法提取大豆油脂及蛋白质的影响研究[J ].[J].食品工业科技,2009,10:240-
[3]陈冬梅,陈明星, 衣海龙等.碱性蛋白酶水解玉米蛋白粉的研究[J ][J].农产品加工,2011,4:70-72
[4]赵丛丛,曾里,宋娜等.碱性蛋白酶提取大米水解蛋白的研究[J].食品与发酵科技,2010,46(1):85-88
[5]刘建龙,王瑞明,刘建军等.酶的固定化技术研究进展[J].中国酿造,2005,9:4-6
[6]袁定重,张秋禹,侯振宇等.固定化酶载体材料的最新研究进展[J].材料导报,2006,20(1):69-72
[7]杨晋青,叶盛权, 郭祀远等.磁性壳聚糖微球的制备及其应用[J].现代食品科技,2008,24(10):1079-1082
[8]刘静娜,黄印强,钱垚.壳聚糖的制备及其对脂肪酶的固定化研究[J].中国酿造,2010,221(8):-
[9]郭伟利,侯红萍.磁性壳聚糖微球固定化糖化酶研究[J].酿酒科技,2010,189(3):17-19
[10]Xie W L,Ma N.Immobilized lipase on Fe3O4糖nanoparticles as biocatalyst for biodesel production[J].Energy fuels,2009,23:1347-1353
[11]费颖.磁性壳聚糖微球固定化乳糖酶及其在乳果糖制备中的应用[D].无锡;江南大学,200
[12]张海真.磁性壳聚糖微球的表面改性表征及吸附行为研究[D].武汉;中南民族大学,200
[13]Ying W.Luminescent CdTe and CdSe semiconductor nanocrystals:preparation,optical properties and applicationsJournal of Nanoscience and Nanotechnology[J].,2008,8:1068-1091
[14]龙胜亚.磁性壳聚糖微球的制备及其在漆酶固定化中的应用[D].武汉;武汉理工大学,200
[15]Liu X Q,Guan Y P,Shen R,et al.Immobilization of lipase onto micron size magnetic beads[J].J Chromatogr B,2005,822:91-97
[16]李桂银.磁性纳米壳聚糖微球的制备及其固定化酵母细胞的研究[D].南京;中南大学,200
[17]Yang W.,Peters JI.,Williams R.O.Inhaled nanoparticles-A current review.International Journal of pharmaceutics[J].,2008,356:239-247
[18]谢玮玮.磁性壳聚糖复合材料的制备及性能研究[D].南昌;南昌大学,200
[19]Corr S.A.,Rakovich YP.Gunko Y.K.Multifunctional magnetic-fluorescent nanocomposites for biomedical applications.Nanoscale Research Letters[J].,2008,3:87-104