Kinetic and Stability Improvement of Immobilized Pepper Chitosanase on Chitin by Covalent Bond
Journal Title: Saudi Journal of Medical and Pharmaceutical Sciences - Year 2017, Vol 3, Issue 9
Abstract
Abstract:Kinetic properties and stability studies of pepper chitosanase immobilized on chitin by covalent bond for production of chitooligosaccharides was the main objective of this work. A comparison with the properties that shown by the free one was also carried out. Maximum enzyme activity was observed at 2.825 and 0.576 mg chitosan /reaction mixture for free chitosanase (FC) and immobilized chitosanase (IC), respectively. They followed Michaelis-Menten Kinetics. The values of the Michaelis-Menten constant indicated that IC exhibited higher affinity toward chitosan than that of the free one. Km value was 35.71 mg/reaction mixture for the IC and 17.86 mg/reaction mixture for the free one. The maximum velocities (Vmax) were 12.5 and 47.62 U/mg for IC and FC, respectively. The optimum pH of IC was slightly shifted to acidic range (pH 5.6) from 5.8, which is the optimum for free one. IC exhibited high activity at wide range of temperature from 40 to 60ºC with optimum at 55ºC, while high activity of FC was found at range from 40 to 55ºC with optimum at 45ºC. The thermal and operational stabilities of both free and immobilized chitosanase were also evaluated. Results showed that the immobilization enhanced the enzyme stability. IC showed better thermal stability than the free one. It lost 5-10 % of its original activity after heating at 50oC for 1h, while FC lost 35 % after the same treatment. IC also lost 58% of its original activity after heating at 60oC for 1h, while free one lost 80 % after the same treatment. Results showed 22% rate of hydrolysis after 1 h and reached about 24 and 30 % after 2 and 24 h of hydrolysis. The end product of chitosan hydrolysis by the IC showed N-acetyl glucosamine and mixture of DP 2-6 oligomers. This improvement of the IC properties made it a potential candidate for future use in industrial application especially in chitooligosaccharides production. Keywords:Chitosanase (EC 3.2.1.132), Pepper (Capsicum annuum) leaves, kinetic properties, operational and thermal stability
Authors and Affiliations
Sanaa T El-Sayed, Nagwa I Omar, El-Sayed M El-Sayed, Wafaa G Shousha
Keywords
Related Articles
- EP ID EP391801
- DOI -
- Views 71
- Downloads 0
Formulation and Evaluation of Colon Targeted pH Dependent Microcapsules of Thymoquinone for Colorectal Cancer
Abstract:Aim of the present work was to prepare the colon-targeting pH dependent microcapsules of Thymoquinone for the treatment of colorectal cancer. pH dependent polymer Eudragit L100,and S100 were used to formulate th...
Identification of Cancer Marker Vinculin from Caco-2 Cancer Cell By Using UPLC/MS Method
Identification of Cancer Marker Vinculin from Caco-2 Cancer Cell By Using UPLC/MS Method Abstract:Vinculin is one the most common proteins that have been studied commonly in recent years.Vinculin has anti-tumor effect an...
Evaluation of Adaptogenic Activity of Anacyclus pyrethrum L.in Animal Models
Abstract:Anacyclus pyrethrum L. root is one among the ingredients of polyherbal formulation frequently employed for the treatment of stress by Ayurvedic practioners. So, the objective of this study was to scientifically...
To Study Effect of Transversus Abdominis Plane Block Using 0.25% Levobupivacaine on Post-Operative Analgesia in Patients Undergoing Open Prostectomy
Patients undergoing surgery are most feared of postoperative pain. All the techniques for multimodal analgesia has some or the other side effects. Transversus abdominis plane (TAP) block is newer, effective and comparati...
A Review on the Prevention of Aminoglycosides Induced Ototoxicity
Abstract:Aminoglycosides antibiotics have been in use for a long time. They are bactericidal in nature and are used to treat infections by Gram-negative bacteria. Studies have revealed their dangerous effect on human bod...