Production, Purification and Characterization of Polygalacturonase from Bacillus licheniformis SHG10
Shaban, N., Ghonaim, T., Masoud, A., Eltokhy, N., Embaby, A. (2017). Production, Purification and Characterization of Polygalacturonase from Bacillus licheniformis SHG10. EKB Journal Management System, 13(1), 95-109. doi: 10.21608/blj.2017.47599
Nadia Z. Shaban; Tayssir M. Ghonaim; Aliaa A. Masoud; Nabil Eltokhy; Amira M. Embaby. "Production, Purification and Characterization of Polygalacturonase from Bacillus licheniformis SHG10". EKB Journal Management System, 13, 1, 2017, 95-109. doi: 10.21608/blj.2017.47599
Shaban, N., Ghonaim, T., Masoud, A., Eltokhy, N., Embaby, A. (2017). 'Production, Purification and Characterization of Polygalacturonase from Bacillus licheniformis SHG10', EKB Journal Management System, 13(1), pp. 95-109. doi: 10.21608/blj.2017.47599
Shaban, N., Ghonaim, T., Masoud, A., Eltokhy, N., Embaby, A. Production, Purification and Characterization of Polygalacturonase from Bacillus licheniformis SHG10. EKB Journal Management System, 2017; 13(1): 95-109. doi: 10.21608/blj.2017.47599

Production, Purification and Characterization of Polygalacturonase from Bacillus licheniformis SHG10

Biochemistry Letters
Article 9, Volume 13, Issue 1, 2017, Page 95-109  XML PDF (928.88 K)
Document Type: Original Article
DOI: 10.21608/blj.2017.47599
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Authors
Nadia Z. Shaban1; Tayssir M. Ghonaim1; Aliaa A. Masoud1; Nabil Eltokhy2; Amira M. Embaby3
1Department of Biochemistry, Faculty of Science, Alexandria University
2City of Scientific Researches and Technological Applications, Borg El Arab, Alexandria, Egypt
3Department of Biotechnology, Institute of Graduate studies and Research, Alexandria University
Abstract
 




 Background: Microorganisms are the best source of pectinolytic enzymes as they allow an economical technology with low resource consumption. Objectives: The present study was carried out to isolate, purify and characterize polygalacturonase (PGase) form Bacillus licheniformis SHG10. Methods: The concentrated dialysed cell free extract was loaded on prepacked DEAE-Sepharose Fast-Flow ion exchange chromatography. The active PGase fractions were concentrated and loaded again on sephacryl Fast-Flow High Resolution (FF S-100 HR) chromatography. Molecular weight was determined using slab-gel SDS-Polyacrylamid gel electrophoresis and Characterization of the purified enzyme was performed. Results: The results showed that the enzyme was purified with a purification fold (33.34) and yield (41.73%) with specific activity (8.67 μ moles/min/mg protein). It has a molecular mass of about 68.0 kDa. While, on SDS-PAGE electrophoresis, the purified PGase appeared as single band with molecular mass of about 34.0 kDa, suggesting that the purified PGase is a dimer protein. The purified enzyme exhibited maximal activity at a temperature of 45oC and pH 8.5. The maximum velocity of the enzyme in presence of citrus pectin and pectate as substrates were 10.98 and 14.7 μ mol galacturonate/min/mg protein, respectively. The Michaelis constant (Km) values were 0.085 and 0.039 mM, respectively, indicating that the purified PGase has higher affinity to pectate (non-methylated pectic substance) than citrus pectin as substrate. 
Keywords
Polygalacturonase; Characterization; Pectinolytic enzymes; Bacillus licheniformis SHG10
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