Open AccessPartial Purification and Characterisation of Pectinase Produced by Aspergillus niger LFP-1 Grown on Pomelo Peels as a Substrate
Author(s) -
Mohd Taufiq Mat Jalil,
İbrahim Demirtaş
Publication year - 2021
Publication title -
tropical life sciences research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.313
H-Index - 16
eISSN - 2180-4249
pISSN - 1985-3718
DOI - 10.21315/tlsr2021.32.1.1
Subject(s) - pectinase , aspergillus niger , chromatography , chemistry , ammonium sulfate precipitation , sephadex , solid state fermentation , size exclusion chromatography , pectin , ammonium sulfate , molecular mass , fermentation , substrate (aquarium) , amberlite , ammonium , yield (engineering) , ion chromatography , enzyme , biochemistry , biology , organic chemistry , materials science , ecology , adsorption , metallurgy
In the present study, pectinase was produced by local fungal isolate, Aspergillus niger LFP-1 grown on pomelo peels as a sole carbon source under solid-state fermentation (SSF). The purification process begins with the concentration of crude enzyme using ammonium sulfate precipitation and followed by purification using anion-exchange column chromatography (DEAE-Sephadex) and subsequently using gel filtration column chromatography (Sephadex G-100). On the other hand, the molecular weight of the purified enzyme was determined through SDS-PAGE. The findings revealed the crude enzyme was purified up to 75.89 folds with a specific activity of 61.54 U/mg and the final yield obtained was 0.01%. The molecular mass of the purified pectinase was 48 kDa. The optimum pH and temperature were 3.5 and 50°C, respectively. This enzyme was stable at a range of pH 3.5 to 4.5 and a relatively high temperature (40°C–50°C) for 100 min. The K m and V max were found to be 3.89 mg/mL and 1701 U/mg, respectively. Meanwhile, pectin from citrus fruit and the metal ion (Co 2+ ) were the best substrate and inducer to enhance pectinase yield, respectively.