Malaysian Journal of Analytical Sciences Vol 20 No 4 (2016): 799 - 805

DOI: http://dx.doi.org/10.17576/mjas-2016-2004-12

 

 

 

ISOLATION AND PURIFICATION OF LYSOZYME FROM ALBUMIN: EFFECT OF ALBUMIN CONCENTRATION, pH AND IONIC STRENGTH OF BUFFER SOLUTION

 

(Pemisahan dan Penulenan Lisozim dari Albumin: Kesan Kepekatan Albumin, pH dan Kekuatan Ionik Larutan Penimbal)

 

Amirah Hamzah¹, Sofiah Hamzah¹*, Fatin Mohd Nasir¹, and 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

The low content of lysozyme in albumin makes its purification process becomes complicated and challenge either in lab or industrial scale system. This study aimed to investigate the parameters influencing the purification performance lysozyme from chicken egg white namely initial concentration of albumin, pH and ionic strength of buffer solution. Immobilized metal affinity chromatography (IMAC) beads were prepared using chitosan-coated silica beads which then crosslinked with glutaraldehyde (GTA) and reacted with metal ion copper (Cu) for metal ion immobilization to be used as purification tools. The prepared beads were characterized in term of morphology and structure using scanning electron microscope (SEM). Column chromatographic has been utilized for evaluation performance of IMAC for lysozyme separation. Optimum recovery obtained using 20 mg/ml CEW concentration at pH 7, with 0.05 M ionic strength. The finding of this study exhibited a good pathway to design an affinity system for lysozyme purification either from chicken egg white or other sources in the future.

 

Keywords: lysozyme, chicken egg white, purification, pH, ionic strength, affinity chromatography

 

Abstrak

Kandungan lisozim yang rendah di dalam albumin membuatkan proses penulenan menjadi semakin rumit dan mencabar sama ada sistem di makmal atau industri. Kajian ini bertujuan untuk mengoptimumkan parameter yang mempengaruhi prestasi penulenan lisozim dari albumin iaitu kepekatan awal albumin, pH dan kekuatan ionik larutan penimbal. Kromatografi affiniti logam pegun telah disediakan dengan menggunakan silika yang bersalut kitosan yang kemudiannya diaktifkan menggunakan glutaraldehid (GTA) serta bertindak balas dengan ion logam kuprum bagi pemegunan ion logam (kuprum) untuk digunakan sebagai alat penulenan. Pencirian silika telah dilakukan dari segi morfologi dan struktur dengan menggunakan mikroskop elektron pengimbas (SEM). Kolum kromatografi digunakan untuk menilai prestasi IMAC bagi pemisahan lisozim. Penghasilan yang optimum diperolehi dengan menggunakan 20 mg/ml kepekatan putih telur pada pH 7 dengan kekuatan ionik sebanyak 0.05 M. Hasil kajian ini menunjukkan cara yang baik untuk mereka bentuk sistem affiniti untuk penulenan lisozim sama ada daripada sumber putih telur atau sumber lain pada masa akan datang.

 

Kata kunci: lisozim, albumin, penulenan, pH, kekuatan ionik, kromatografi affiniti

 

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