Partial Purification and Characterisation of Pectinase Produced by Aspergillus niger LFP-1 Grown on Pomelo Peels as a Substrate

Area of Science:

• Biotechnology
• Enzyme Technology
• Industrial Microbiology

Background:

• Pectinase enzymes are crucial for various industrial applications, including food processing and biofuel production.
• Utilizing agricultural waste like pomelo peels for enzyme production offers a sustainable approach.

Purpose of the Study:

• To produce and purify pectinase from a local fungal isolate, Aspergillus niger LFP-1.
• To characterize the biochemical properties and stability of the purified pectinase.
• To identify optimal conditions and substrates for enhanced pectinase production.

Main Methods:

• Solid-state fermentation (SSF) using Aspergillus niger LFP-1 on pomelo peels.
• Enzyme purification involving ammonium sulfate precipitation, DEAE-Sephadex, and Sephadex G-100 chromatography.
• Enzyme characterization including SDS-PAGE, optimal pH/temperature determination, stability assays, and kinetic analysis (Km, Vmax).

Main Results:

• Pectinase was purified 75.89-fold with a specific activity of 61.54 U/mg and a yield of 0.01%.
• The purified enzyme has a molecular mass of 48 kDa, with optimal activity at pH 3.5 and 50°C.
• The enzyme demonstrated stability in a pH range of 3.5-4.5 and temperatures of 40-50°C for 100 min, with Km of 3.89 mg/mL and Vmax of 1701 U/mg.
• Citrus fruit pectin and Cobalt ions (Co2+) were identified as the optimal substrate and inducer, respectively.

Conclusions:

• Aspergillus niger LFP-1 is a suitable source for producing industrially relevant pectinase.
• The characterized pectinase exhibits favorable properties for potential applications in biotechnological processes.
• Pomelo peels serve as an effective substrate for SSF-based pectinase production, highlighting sustainable bioprocessing.