Liqaa Y. Mohsen, Jawad K. A. Al-Janabi, Mohammad A. Jebor
Abstract: These experiments were conducted to study the production, purification, characterization, and molecular identification of Polygalacturonase enzyme from F. oxysporum and F. sacchari were investigated. The results showed that the optimum incubation period for Polygalacturonase in F. oxysporum and F. sacchari were occurred within two days. Polygalacturonase exhibited maximal activity at pH 6, 27C and 0.5 % pectin concentration. The results revealed that the best ratio for exo Polygalacturonase precipitation for F. oxysporum and F. sacchari was 70 and 90 % of ammonium sulphate respectively. Two peaks of PG were appeared in ion exchange chromatography purified from F.oxysporum. While in case of F. sacchari, only three peaks of protein and one peak of enzyme activity were shown The results showed that the molecular weight of Polygalacturonase from F. oxysporum (two peaks) using SDS-PAGE were approximately 53KD, and about 41 KD for PG purified from F.sacchari under denaturation conditions. For characterization of Polygalacturonase purified from F.oxysporum, the results reported the highest activity of PG (A) occurred at pH 5 while peak (B) was showed highest activity at pH 6. On other hand, the maximum enzyme activity of Polygalacturonase in F. sacchari was obtained at pH 5. Best growth (two peaks) achieved at 40C for F. oxysporum and 45 C for F. sacchari respectively. Finally, molecular identification of exo Polygalacturonase gene in F. oxysporum and F. sacchari was investigated using PCR technique. The PCR primers designed from the conserved region of known fungal produced 1700 bp product from the genomic DNA of F.oxysporum and 1200 bp product from the genomic DNA of F.sacchari.
Keywords: Purification, polygalacturonase enzyme, molecular identification of pgx1 gene