Identification and characterization of thermostable endo‐polygalacturonase II B from Aspergillus luchuensis

Endo-polygalacturonase II B (PgaB) from Aspergillus luchuensis was orthologous to endo-polygalacturonase from Aspergillus niger with mutant sites Thr42Ser and Glu52Ala. Mature pgaB gene was cloned from the genomic DNA of A. luchuensis and secreted expressed with over 90% purity in Pichia Pastoris and reached 1.0 g/L after 144 hr culture. The recombinant PgaB was further purified by Ni-NTA chromatography. Using polygalacturonic acid (PGA) as substrate, the optimal condition for PgaB activity was 40 degrees C and pH 4.5, respectively. Km and Vmax of PgaB were 0.19 mmol/l and 103.58 mumol min(-1) mg(-1) , respectively. The relative activity of PgaB remained more than 60% and 40% of maximum activity at 50 and 60 degrees C for 7 hr. PgaB increased the light transmittance by 85% and showed high efficiency in juice clarification. The main product was galacturonic acid oligosaccharides with degrees of polymers (DP) 1-3. The PgaB is a potential pectinolytic enzyme in food industries. PRACTICAL APPLICATIONS: Endo-polygalacturonase II B (PgaB) was identified from Aspergillus luchuensis, a filamentous fungus widely used in food and beverage fermentation in East Asia. PgaB still kept its most activity at 60 degrees C for 7 hr. Polygalacturonic acid (PGA) can be digested effectively by the PgaB and the main products are galacturonic acid oligosaccharides with degrees of polymers (DP) 1-3. PgaB shows high efficiency in juice clarification. The PgaB is a potential pectinolytic enzyme for the applications in food industries.