Malaysian
Journal of Analytical Sciences Vol 20 No 3 (2016): 578 - 584
DOI:
http://dx.doi.org/10.17576/mjas-2016-2003-17
IMMOBILISED METAL AFFINITY CHROMATOGRAPHY (IMAC)
BEADS FOR LYSOZYME SEPARATION: SYNTHESIS AND CHARACTERIZATION STUDY
(Kromatografi Afiniti Logam Dipegun (IMAC) untuk
Pemisahan Lysozim: Sintesis dan Kajian Pencirian)
Fatin Mohd Nasir1, Sofiah Hamzah1*, Amirah Hamzah1,
Nora’aini Ali1, Marinah Mohd Ariffin2
1School of Ocean Engineering
2School of Marine Science and Environment
Universiti Malaysia Terengganu, 21030 Kuala
Terengganu, Terengganu, Malaysia
*Corresponding author: sofiah@umt.edu.my
Received: 24
February 2015; Accepted: 27 October 2015
Abstract
Immobilized
metal affinity chromatography (IMAC) has been established as a highly specific chromatographic
technique for the production of enzymes and proteins including lysozyme. This
study aimed to prepare and characterize the IMAC beads for lysozyme separation.
Silica gel served as chromatographic matrix which has been coated with chitosan
layer and crosslinked with glutaraldehyde (GTA-CTS-SiO2) since it is
very convenient to promote fixation. Various IMAC ligands were immobilized by
chelating 1500 mg/l Cu2+, Zn2+, Fe2+, Fe3+
and Al3+ ions, respectively on GTA-CTS-SiO2. IMAC which
was immobilized with 1200mg/l Cu2+ exhibited the maximal immobilization
capacity (27.08mg/g) of within 30 minutes incubation time. This fundamental
study can be a momentous pathway to develop an efficient chromatographic system
for lysozyme separation in the future.
Keywords: silica gel, chitosan, glutaraldehyde, metal ions,
copper (II) ion
Abstrak
Kromatografi afiniti logam dipegun (IMAC) telah
diperlihatkan sebagai satu teknik kromatografi yang sangat spesifik untuk
pengeluaran enzim dan protein termasuk lysozim. Kajian ini bertujuan untuk
menyediakan dan mencirikan manik IMAC bagi tujuan pemisahan lysozim. Gel silika
berperanan sebagai sokongan kepada kromatografi yang telah dilapisi dengan
lapisan kitosan dan diaktifkan dengan glutaraldehid (GTA-CTS-SiO2)
kerana kesesuaiannya yang berupaya menggalakkan lekatan. Beberapa ligan IMAC dengan
kepekatan 1500mg/l (Cu2+, Zn2+, Fe2+, Fe3+
dan Al3+ ion) telah dipegunkan pada
Kata kunci: gel silika, kitosan, glutaraldehid, ion logam, ion kuprum (II)
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