Immobilization of L-asparaginase on aspartic acid functionalized graphene oxide nanosheet: Enzyme kinetics and stability studies

被引:85
作者
Monajati, Maryam [1 ,2 ]
Borandeh, Sedigheh [1 ]
Hesami, Anahita [1 ]
Mansouri, Dorsa [1 ]
Tamaddon, Ali Mohammad [1 ,3 ]
机构
[1] Shiraz Univ Med Sci, Ctr Nanotechnol Drug Delivery, Shiraz, Iran
[2] Univ Tehran Med Sci, Dept Nanopharmaceut, Fac Pharm, Tehran, Iran
[3] Shiraz Univ Med Sci, Sch Pharm, Dept Pharmaceut, Shiraz, Iran
关键词
Enzyme immobilization; Nanocomposite; Graphene oxide; L-Asparaginase; COVALENT IMMOBILIZATION; BIOMEDICAL APPLICATIONS; BETA-AMYLASE; NANOPARTICLES; DELIVERY; NANOBIOCATALYST; DERIVATIVES;
D O I
10.1016/j.cej.2018.08.058
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
There is an increasing interest in using L-asparaginase in medical fields and food processing industries. Enzyme immobilization is an attractive field to improve L-asparaginase activity and stability. Graphene oxide (GO) is a promising candidate for enzyme immobilization due to its large specific surface area. In this study, GO was first functionalized with L-aspartic acid (GO-Asp), and then L-asparaginase was immobilized on the GO-Asp either physically or through chemical conjugation. A significant enzyme loading was achieved through covalent immobilization (100% immobilization efficiency). Stability of free and the immobilized L-asparaginase was examined at various temperatures (20-60 degrees C) and pH (5-9). The covalently immobilized L-asparaginase showed higher enzyme activity than free enzyme at pH 8 with the maximum recovered activity of 100%, 90.5% and 40.6% after 24 h of incubation at 20 degrees C, 40 degrees C and 60 degrees C, respectively. In addition, the covalently immobilized L-asparaginase on GO-Asp showed 42% recovered activity after eight continuous reaction cycles at 60 degrees C. The kinetic parameters of the immobilized and free enzyme were also calculated, indicating no significant changes in the enzyme affinity through covalent conjugation. The results clearly reflect the suitability of GO-Asp as a nanosheet support for L-asparaginase loading as well as its usage in future industrial applications.
引用
收藏
页码:1153 / 1163
页数:11
相关论文
共 60 条
[1]   Improving Interfacial Interaction of L-Phenylalanine-Functionalized Graphene Nanofiller and Poly(vinyl alcohol) Nanocomposites for Obtaining Significant Membrane Properties: Morphology, Thermal, and Mechanical Studies [J].
Abdolmaleki, Amir ;
Mallakpour, Shadpour ;
Borandeh, Sedigheh .
POLYMER COMPOSITES, 2016, 37 (06) :1924-1935
[2]  
Ahmad A., 2013, INT J PHARM BIOSCIEN, P274
[3]   Nanostructure L-asparaginase-fatty acid bioconjugate: Synthesis, preformulation study and biological assessment [J].
Ashrafi, Hajar ;
Amini, Mohsen ;
Mohammadi-Samani, Soliman ;
Ghasemi, Younes ;
Azadi, Amir ;
Tabandeh, Mohammad Reza ;
Kamali-Sarvestani, Eskandar ;
Daneshamouz, Saeid .
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 2013, 62 :180-187
[4]   Nanodevices for the immobilization of therapeutic enzymes [J].
Bosio, Valeria E. ;
Islan, German A. ;
Martinez, Yanina N. ;
Duran, Nelson ;
Castro, Guillermo R. .
CRITICAL REVIEWS IN BIOTECHNOLOGY, 2016, 36 (03) :447-464
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   Catalytic Oxidation of Phenol and 2,4-Dichlorophenol by Using Horseradish Peroxidase Immobilized on Graphene Oxide/Fe3O4 [J].
Chang, Qing ;
Huang, Jia ;
Ding, Yaobin ;
Tang, Heqing .
MOLECULES, 2016, 21 (08)
[7]   Covalent immobilization of peanut β-amylase for producing industrial nano-biocatalysts: A comparative study of kinetics, stability and reusability of the immobilized enzyme [J].
Das, Ranjana ;
Talat, Mahe ;
Srivastava, O. N. ;
Kayastha, Arvind M. .
FOOD CHEMISTRY, 2018, 245 :488-499
[8]   Covalent immobilization of β-amylase onto functionalized molybdenum sulfide nanosheets, its kinetics and stability studies: A gateway to boost enzyme application [J].
Das, Ranjana ;
Mishra, Himanshu ;
Srivastava, Anchal ;
Kayastha, Arvind M. .
CHEMICAL ENGINEERING JOURNAL, 2017, 328 :215-227
[9]   Immobilization on graphene oxide improves the thermal stability and bioconversion efficiency of D-psicose 3-epimerase for rare sugar production [J].
Dedania, Samir R. ;
Patel, Manisha J. ;
Patel, Dijit M. ;
Akhani, Rekha C. ;
Patel, Darshan H. .
ENZYME AND MICROBIAL TECHNOLOGY, 2017, 107 :49-56
[10]   The chemistry of graphene oxide [J].
Dreyer, Daniel R. ;
Park, Sungjin ;
Bielawski, Christopher W. ;
Ruoff, Rodney S. .
CHEMICAL SOCIETY REVIEWS, 2010, 39 (01) :228-240