Muinat O Kazeem
UPM, Malaysia
Title: Partitioning and purification of carboxymethyl cellulase from in aqueous biphasic system extraction
Biography
Biography: Muinat O Kazeem
Abstract
Statement of problem: Caboxymethyl cellulase (CMC) is one of the significant enzyme in biofuel, textile, paper, brewing and animal feed industries, because of its ability to catalyze enzymatic hydrolysis cellulosic material into simple sugars. However, the downstream processing of CMCase from fermentation broth involves tedious conventional chromatographic steps which is time consuming and complicated. Researchers have reported different methods such as gel filtration and ion-exchange chromatography, ammonium sulphate fractionation and chromatography on DEAE-sephadex A-50 and Sephadex G-100 80 and ultrafiltration. However, application in large scale becomes difficult due to high cost, low productivity and multiple purification steps involved; thus, a cost effective strategy for recovery of CMCase is in demand. One possible method to overcome this challenge is aqueous biphasic system extraction (ABSE). The purpose of this study is to examine the effectiveness of applying ABSE involving polyethylene glycol/salt for the partitioning behavior and recovery of CMCase produced from thermophilic Bacillus licheniformis 2D55 fermentation broth. Methodology: The effect of ABS variables of type of salt, PEG molecular weight, crude loading, pH and additional sodium chloride (NaCl) on partitioning behavior of CMCase were investigated. Findings: A 14 % (w/w) crude load , 1 % NaCl, pH 7.0 produced a CMCase recovery yield (Y) of 88.63 %, a partition coefficient (K) of 0.21 and purification factor (PFT) of 10.3 from the bottom phase of PEG 8000/sodium citrate ABSE. The hydrophobicity nature of the CMCase surface enhanced the partition and extraction of the CMCase to the salt rich bottom sodium citrate phase. Likewise, impurities undesired proteins were separated to the PEG 8000 rich top phase. The effectiveness of the recovery was confirmed by SDS-PAGE analysis. The study suggest that ABSE may be a potential strategy for industrial purification of CMCase form fermentation broth