High throughput covalent immobilization process for improvement of shelf-life,operational cycles,relative activity in organic media and enzymatic kinetics of urease and its application for urea removal from water samples |
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| Institution: | 1. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, 11800, Penang, Malaysia;2. Veterinary Research Institute, 59, Jalan Sultan Azlan Shah, Ipoh, 31400, Perak, Malaysia;3. Usains Biomics Laboratory Testing Services Sdn Bhd., Universiti Sains Malaysia, Minden, 11800, Penang, Malaysia;4. Department of Applied Biology, Kyoto Institute of Technology, Sakyo-ku, Kyoto, 606-8585, Japan;5. School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, 15200, Kelantan, Malaysia;6. RIKEN, Wako, Saitama, 351-0198, Japan;1. Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, Nanjing, 210037, People’s Republic of China;2. College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, People’s Republic of China;3. Jiangsu Province Key Laboratory of Green Biomass-based Fuels and Chemicals, Nanjing, 210037, People’s Republic of China;1. MOE Key Laboratory of Groundwater Circulation and Environmental Evolution, School of Water Resources and Environment, China University of Geosciences (Beijing), Beijing 100083, PR China;2. Beijing Key Laboratory of Water Resources and Environmental Engineering, School of Water Resources and Environment, China University of Geosciences (Beijing), Beijing 100083, PR China;3. Yickxin Technology Development Ltd. Co., Shenzhen 518000, PR China;1. Green Chemistry and Materials Group, Korea Institute of Industrial Technology (KITECH), Cheonan, 31056, South Korea;2. Green Process and System Engineering Major, University of Science and Technology, Daejeon, 34113, South Korea;3. Department of Biochemical Engineering, Gangneung-Wonju National University, Gangneung, 25457, South Korea;1. Brain Institute (Instituto do Cérebro-INSCER), Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, RS, Brazil;2. Center of Biotechnology, Universidade Federal do Rio Grande do Sul Porto Alegre, RS, Brazil |
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| Abstract: | High throughput covalent urease immobilization was performed through the amide bond formation between the urease and the amino-functional MNPs. The enzyme’s performances, including shelf-life, reusability, enzymatic kinetics, and the enzyme relative activity in organic media was improved. At optimal conditions, the immobilization efficiency was calculated about 95.0% with keeping 94.7% of the urease initial specific activity. The optimal pH for maximum activity of the free and immobilized urease was calculated as 7.0 at 37.0 °C and 8.0 at 60.0 °C, respectively. The kinetics studies showed the Km of 26.0 mM and 8.0 mM and the Vmax of 5.31 μmol mg−1 min−1 and 3.93 μmol mg−1 min−1 for the free and immobilized urease, respectively. The ratio Kcat/Km as a measure of catalytic efficiency and enzyme specificity was calculated as 0.09 mg mL−1 min−1 and 0.22 mg mL−1 min−1 for the free and immobilized urease, respectively, indicating an improvement in the enzymatic kinetics. The shelf-life and operational studies of immobilized urease indicated that approximately 97.7% and 88.5% of its initial activity was retained after 40 days and 17 operational cycles, respectively. The immobilized urease was utilized to urea removal from water samples with an efficiency between 91.5–95.0%. |
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| Keywords: | Urease Covalent immobilization Magnetic nanoparticles Operational studies Urea removal |
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