Covalent immobilization of chloroperoxidase onto magnetic beads: Catalytic properties and stability

2008-02-15
Bayramoglu, Guelay
Kiralp, Senem
Yilmaz, Meltem
Toppare, Levent Kamil
Arica, M. Yakup
Show full item record
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
184
views
0
downloads
Amino groups containing magnetic beads were used in covalent immobilization of the enzyme "chloroperoxidase (CPO)" which is one of a few enzymes that can catalyse the peroxide dependent oxidation of a wide spectrum of organic and inorganic compounds. The magnetic poly(glycidylmethacrylate-methylmethacrylate-etbyleneglycol dimethacrylate), magnetic p(GMA-MMA-EGDMA) beads were prepared via suspension polymerization in the presence of ferric ions. The magnetic beads were characterized with scanning electron microscope (SEM), Fourier transform infrared (FTIR), Mossbauer spectroscopy and vibrating sample magnetometer (VSM). The magnetic beads were derivatized sequentially with ammonia and glutaraldehyde, and CPO was covalently immobilized on the support via reaction of the amino groups of the enzyme under mild conditions. The effect of various parameters including pH, temperature and enzyme concentration on the immobilization efficiency of CPO onto glutaric dialdhyde activated magnetic beads was evaluated. Magnetic measurement revealed that the resultant CPO-immobilized magnetic beads were superparamagnetic with a saturation magnetization of 18.2 emu/g. The analysis of FTIR spectra confirmed the binding of CPO on the magnetic beads. The maximum amount of immobilized CPO on the magnetic beads was 2.94 mg/g support. The values of Michaelis constants Km for immobilized CPO was significantly larger, indicating decreased affinity by the enzyme for its substrate, whereas V-max values were smaller for the immobilized CPO. However, the CPO immobilized on the magnetic beads resulted in an increase in enzyme stability with time.
Biotechnology, Environmental Engineering, Bioengineering, Biomedical Engineering
https://hdl.handle.net/11511/37057
BIOCHEMICAL ENGINEERING JOURNAL
Department of Chemistry, Article

Suggestions

Carbon sources affect metabolic capacities of Bacillus species for the production of industrial enzymes: theoretical analyses for serine and neutral proteases and alpha-amylase
Çalık, Pınar (Elsevier BV, 2001-07-01)
The metabolic fluxes through the central carbon pathways were calculated for the genus Bacillus separately for the enzymes serine alkaline protease (SAP), neutral protease (NP) and alpha -amylase (AMY) on five carbon sources that have different reduction degrees (gamma), to determine the theoretical ultimate limits of the production capacities of Bacillus species and to predict the selective substrate for the media design. Glucose (gamma = 4.0), acetate (gamma = 4.0), and the TCA cycle organic-acids succina...
The biodegradation of benzene, toluene and phenol in a two-phase system
Hamed, TA; Bayraktar, E; Mehmetoglu, U; Mehmetoglu, T (Elsevier BV, 2004-07-15)
A two-phase aqueous-organic system was used to degrade benzene, toluene and phenol, individually and as mixture by Pseudomonas putida F1 (ATCC 700007). In the first stage of the work, the effect of the phase ratio (0-1, v/v) and the agitation rate (150 and 200 rpm) on the biodegradation process was investigated in an orbital shaker using 2-undecanone as the solvent and the most suitable phase ratio and agitation rate were found to be 0.0625 (v/v) and 200 rpm, respectively. P. putida F1 was added into the aq...
Analyses of extracellular protein production in Bacillus subtilis - II: Responses of reaction network to oxygen transfer at transcriptional level
KOCABAŞ, PINAR; ÇALIK GARCİA, GÜZİDE; Çalık, Pınar; Ozdamar, Tuncer H. (Elsevier BV, 2017-11-15)
Oxygen transfer influences intracellular fluxes which are orchestrated by genome and its transcription in Bacillus subtilis throughout fermentation in recombinant human growth hormone (rhGH) production. Responses of B. subtilis reaction network to oxygen transfer were analysed at transcriptional level with determined transcriptome and calculated intracellular fluxes by the reconstructed genome scale model iBsu1144(rhGH) based on updated gene-enzyme-reaction data. iBsu1144(rhGH) employing 1067 reactions link...
Biodegradation of 4-CP and 2,4-DCP mixture in a rotating biological contactor (RBC)
şahinkaya, Erkan; Dilek, Filiz Bengü (Elsevier BV, 2006-09-01)
In this study, the performance of a two stage rotating biological contactor (RBC) was evaluated for the treatment of synthetic wastewater containing peptone, 4-chlorophenol (4-CP) and 2,4-dichlorophenol (2,4-DCP) at 5 rpm. Also, the effect of biogenic substrate (peptone) concentration on the reactor performance was investigated. High chlorophenols (> 98%) and COD (> 94%) removals were achieved throughout the reactor operation in the first stage and the second stage behaved as a polishing step. The observed ...
Regulatory effects of alanine-group amino acids on serine alkaline protease production by recombinant Bacillus licheniformis
Çalık, Pınar; Ozdamar, TH (Wiley, 2003-04-01)
Influences of the concentration and addition time of alanine-group amino acids, i.e. alanine, leucine and valine, on serine alkaline protease (SAP) synthesis were investigated by Bacillus licheniformis (DSM 1969) carrying pHV1431::subC in a defined medium to identify the amino acids creating intracellular reaction-rate limitation in SAP production. While the precursors of alanine-group amino acids, pyruvate and alanine, did not affect SAP production considerably within the range 0-15 mM, the addition of leu...
Citation Formats
G. Bayramoglu, S. Kiralp, M. Yilmaz, L. K. Toppare, and M. Y. Arica, “Covalent immobilization of chloroperoxidase onto magnetic beads: Catalytic properties and stability,” BIOCHEMICAL ENGINEERING JOURNAL, pp. 180–188, 2008, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/37057.
METU IIS - Integrated Information Service open@metu.edu.tr