Immobilization of Pullulanase from Local Isolate Bacillus subtilis MF467279 Strain onto Magnetic@TEMPO Cellulose Nanospheres for Hydrolysis of Pullulan
Shehata, A., Abdelraof, M., Hasanin, M. (2025). Immobilization of Pullulanase from Local Isolate Bacillus subtilis MF467279 Strain onto Magnetic@TEMPO Cellulose Nanospheres for Hydrolysis of Pullulan. EKB Journal Management System, 68(6), 417-429. doi: 10.21608/ejchem.2024.318233.10348
Abeer Nasr Shehata; Mohamed Abdelraof; Mohamed Hasanin. "Immobilization of Pullulanase from Local Isolate Bacillus subtilis MF467279 Strain onto Magnetic@TEMPO Cellulose Nanospheres for Hydrolysis of Pullulan". EKB Journal Management System, 68, 6, 2025, 417-429. doi: 10.21608/ejchem.2024.318233.10348
Shehata, A., Abdelraof, M., Hasanin, M. (2025). 'Immobilization of Pullulanase from Local Isolate Bacillus subtilis MF467279 Strain onto Magnetic@TEMPO Cellulose Nanospheres for Hydrolysis of Pullulan', EKB Journal Management System, 68(6), pp. 417-429. doi: 10.21608/ejchem.2024.318233.10348
Shehata, A., Abdelraof, M., Hasanin, M. Immobilization of Pullulanase from Local Isolate Bacillus subtilis MF467279 Strain onto Magnetic@TEMPO Cellulose Nanospheres for Hydrolysis of Pullulan. EKB Journal Management System, 2025; 68(6): 417-429. doi: 10.21608/ejchem.2024.318233.10348

Immobilization of Pullulanase from Local Isolate Bacillus subtilis MF467279 Strain onto Magnetic@TEMPO Cellulose Nanospheres for Hydrolysis of Pullulan

Egyptian Journal of Chemistry
Volume 68, Issue 6, June 2025, Page 417-429  XML PDF (1.08 MB)
Document Type: Original Article
DOI: 10.21608/ejchem.2024.318233.10348
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Authors
Abeer Nasr Shehata email orcid 1; Mohamed Abdelraof2; Mohamed Hasaninorcid 3
1Biochemisrty Department- Biotechnology Research Institute-National Research Centre
2Microbial Chemistry Department Biotechnology research institute
3Cellulose and Paper Department Chemical Industries Research Institute
Abstract
Bacillus subtilis MF467279 was isolated from the desert plant species Salsola imbricate, that collected from Tur Sinai, Egypt, for production of pullulanase using growth on optimized liquid medium. Pullulanase was partially purified using fractionation by using 20-80% ammonium sulfate with 85% yield and 4.47 purification fold. The partially purified pullulanase was immobilized onto a biopolymer carrier (magnetic@TEMPO cellulose nanospheres) with 90% immobilization efficiency and 85 % immobilization yield. Kienetic parameters (Km and Vmax) for pullulan, soluble starch and amylopectin was achieved. The optimal temperature for free and immobilized enzymes was 40 and 60°C respectively with broad range (4-10) of pH activity for immobilized pullulanase. Characterizations revealed that immobilization highly improved thermostability, reusability and shelf stability. The free and immobilized pullulanse were able to hydrolyze soluble starch, amylopectin glycogen, dextrin and dextran. Studying the end products resulted from hydrolysis of pullulan and starch by free and immobilized enzyme via thin layer chromatography (TLC) indicated that Bacillus subtilis enzyme is pullulanase hydrolase type III which consider the most important hydrolase in starch saccharification process.



Key words. Pullulanase - Bacillus subtilis MF46727- Immobilization- magnetic@TEMPO cellulose nanospheres- Thin Layer chromatography (TLC).
Keywords
Key words. Pullulanase; Bacillus subtilis MF46727; Immobilization; magnetic@TEMPO cellulose nanospheres; Thin Layer chromatography (TLC)
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