Malaysian Journal of Analytical Sciences Vol 26 No 1 (2022): 16 - 28

 

 

 

 

ENHANCING TRYPSIN RECOVERY USING POLYMER-BASED AFFINITY ULTRAFILTRATION MEMBRANE: EFFECTS OF ELUTION pH AND DISPLACING SALTS

 

(Meningkatkan Perolehan Tripsin menggunakan Membran Afiniti Ultrafiltrasi berasaskan Polimer: Kesan pH Pengeluatan dan Penggantian Garam)

 

Norhafiza Ilyana Yatim1, Sofiah Hamzah2*, Maslinda Alias2, Nora'aini Ali1,2, Marinah Mohd Ariffin3, Abdul Wahab Mohammad4

 

1Higher Institution Centre of Excellence (HICoE), Institute of Tropical Aquaculture and Fisheries

2Faculty of Ocean Engineering Technology and Informatics

3Faculty of Science and Marine Environment

Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia

4Faculty of Engineering and Built Environment

Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

 

*Corresponding author: sofiah@umt.edu.my

 

 

Received: 27 August 2021; Accepted: 11 January 2022; Published: 25 February 2022

 

 

Abstract

Downstream processing of trypsin synthesis, which includes purification, is a major issue due to high complexity of the bio-suspension itself and the stress sensitivity of the desired target molecules. Therefore, an affinity membrane was fabricated in this study using polysulfone polymers and modified by chitosan to enhance biofouling resistance. To determine the optimum conditions for maximum trypsin adsorption and desorption during affinity ultrafiltration process, elution buffers and pH were investigated using various types of displacing salts (potassium chloride, KCl; sodium chloride, NaCl; magnesium chloride, MgCl2; calcium chloride, CaCl2) and elution pH (pH 4, 5, 6, 7, 8). The result showed that the buffer using KCl was identified as the best displacing salt as it recovered the highest trypsin of 78.84%, with a purification fold of 1.31. Trypsin recovery increased to 92% and 1.20 purification fold when the experiments were at pH 8. These buffers dissolved the interference chemical bonds comprising trypsin-trypsin inhibitor interactions and restore target trypsin to the permeate stream in an active state for maximum trypsin recovery. The information provided in this study represents a possible future avenue for developing an affinity membrane system.

 

Keywords: affinity, ultrafiltration, trypsin, elution pH, displacing salt creator

 

Abstrak

Pemprosesan hiliran sintesis tripsin, yang termasuk penulinan, adalah isu utama kerana kerumitan bio-pengampaian itu sendiri dan kepekaan tekanan molekul sasaran yang dikehendaki. Oleh itu, membran afiniti telah direka dalam kajian ini menggunakan polimer polisulfon dan diubahsuai oleh kitosan untuk meningkatkan rintangan bio-kotoran. Untuk menentukan keadaan optimum untuk penjerapan tripsin maksimum dan penyahjerapan semasa proses ultrafiltrasi perkaitan, penimbal pengeluat dan pH telah disiasat menggunakan pelbagai jenis garam pengganti (kalium klorida, KCl; natrium klorida, NaCl; magnesium klorida, MgCl2; kalsium klorida, CaCl2) dan pH pengeluatan (pH 4, 5, 6, 7, 8). Hasilnya menunjukkan bahawa penimbal menggunakan KCl dikenal pasti sebagai garam pengganti terbaik kerana ia perolehan tripsin tertinggi sebanyak 78.84%, dengan lipatan penulenan 1.31. Perolehan tripsin meningkat kepada 92% dan 1.20 lipatan penulenan apabila eksperimen berada di pH 8. Penimbal ini telah melarutkan ikatan kimia yang menganggu yang terdiri daripada interaksi perencat tripsin-tripsin dan memulihkan tripsin sasaran ke aliran menelap dalam keadaan aktif untuk perolehan maksimum tripsin. Maklumat yang disediakan dalam kajian ini mewakili jalan masa depan yang mungkin untuk membangunkan sistem membran afiniti.

 

Kata kunci: afiniti, ultrafiltrasi, tripsin, pH pengeluat, pencipta pengganti garam

 

 


Graphical Abstract




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