Article

Optimization of Matrix Components for Improved Catalytic Activities of Cellulase Immobilized on Biochar-Chitosan Beads

Downloads

Chidi Evans, E., Omotayo Omonije, O., & Poritmwa Gontul, I. (2024). Optimization of Matrix Components for Improved Catalytic Activities of Cellulase Immobilized on Biochar-Chitosan Beads. New Energy Exploitation and Application, 3(1), 130–142. https://doi.org/10.54963/neea.v3i1.249

Authors

  • Egwim Chidi Evans 1 Biochemistry Department, Federal University of Technology, Minna, 920101 Niger, Nigeria; 2 Chemistry/Biochemistry Department, Caleb University, Imota-ikorodu 104101, Lagos, Nigeria
  • Oluyemisi Omotayo Omonije
    Biochemistry Department, Federal University of Technology, Minna, 920101 Niger, Nigeria
  • Isaac Poritmwa Gontul Biochemistry Department, Federal University of Technology, Minna, 920101 Niger, Nigeria

Bioethanol is a renewable energy that is gaining popularity globally. It’s biochemical production requires the use of enzyme, especially cellulase. Cellulase is an enzyme that catalyzes the degradation of cellulose and related polysaccharides which finds applications in food, textiles, detergents, biofuels etc. However, the worldwide use of cellulase is limited by its relatively high production costs and low biological activity. This study was design to locally produce biochar-chitosan beads at optimized conditions to immobilize cellulase for improved thermal and storage stability as well as ensure reusability of the enzyme so as to improve biological activity and avoid the continuous production of free cellulase thereby reducing the production cost. Biochar was produced by pyrolyzing sugarcane bagasse in a local airtight chamber for 1 hour. Beads were formed from different ratios of biochar and chitosan in varying concentrations of calcium chloride solution as generated by design expert software version 13. The beads were dried in an oven at 50 0C for 24 hours and functionalized in 25% glutaraldehyde (GDA). The beads were loaded with enzyme (10.06 µmole/min/mL) at room temperature (27 ± 3 oC). Enzyme activity, thermal stability, storage stability and reusability tests were carried out according to standard procedures. The half-life and activation energy were also evaluated. The result showed that the optimum activity of the loaded enzyme (2.63 µmole/min/mL) was obtained when 2.46 g of porous biochar was mixed with 2.48 g chitosan in 5 % Calcium chloride aqueous solution. The immobilized enzyme was able to maintain thermal stability between 30 oC and 70 oC while the activity for free enzyme started declining after 50 oC. Also, the activation energy for immobilized cellulase enzyme (23.17 KJ/mol) was lower than the activation energy (55.146 KJ/mol) for free cellulase. The half-life, when stored at ambient Temperature (27 ± 3 oC), for free enzyme was 0.4 days while the half-life for immobilized enzyme was 3.59 days. Therefore, cellulase immobilized on support locally produced at optimal conditions had improved catalytic properties when compared to the free enzyme. Hence, more indigenous materials and practices may be employed for a cost effective and cheaper industrial processes.

Keywords:

cellulase sugarcane bagasse pyrolysis local chamber composite beads

References

  1. Thapa, S.; Mishra, J.; Arora, N.; Mishra, P.; Li, H.; O’Hair, J.; Bhatti, S.; Zhou, S. Microbial Cellulolytic Enzymes: Diversity and Biotechnology with Reference to Lignocellulosic Biomass Degradation. Rev. Environ. Sci. Biotechnol. 2020, 19, 621–648.
  2. Barbosa, F.C.; Silvello, M.A.; Goldbeck, R. Cellulase and Oxidative Enzymes: New Approaches, Challenges and Perspectives on Cellulose Degradation for Bioethanol Production. Biotechnol. Lett. 2020, 42, 875–884.
  3. Zoghlami, A.; Paës, G. Lignocellulosic Biomass: Understanding Recalcitrance and Predicting Hydrolysis. Front. Chem. 2019, 7, 874.
  4. Mo, H.; Qiu, J.;Yang, C.; Zang, L.; Sakai, E.; Chen, J. Porous Biochar/Chitosan Composites for High Performance Cellulase Immobilization by Glutaraldehyde. Enzyme Microb. Technol. 2020, 138, 109561.
  5. Utomo, Y.; Yuniawati, N.; Wonorahardo, S.; Sumari; Santoso, A.; Kartika K. I.; Susanti, E. Preliminary Study of Immobilized of Cellulase in Silica from the Rice Husk Ash to Hydrolysis Sugarcane Bagasse. In Proceedings of the IOP Conference Series: Earth and Environmental Science, International Conference on Life Sciences and Technology: Malang, Indonesia, 4 September 2018.
  6. Sirisha, V.L.; Jain, A.; Jain, A. Enzyme Immobilization: An Overview on Methods, Support Material, and Applications of Immobilized Enzymes. Adv. Food Nutr. Res. 2016, 79, 179–211.
  7. Smith, P. Soil Carbon Sequestration and Biochar as Negative Emission Technologies. Global Change Biol. 2016, 22, 1315–1324.
  8. Nawaz, M.A.; Rehman, H.U.; Bibi, Z.; Aman, A.; Ul Qader, S.A. Continuous Degradation of Maltose by Enzyme Entrapment Technology Using Calcium Alginate Beads as a Matrix. Biochem. Biophys. Rep. 2015, 4, 250–256.
  9. Tuan Mohamood, N.F.A.Z.; Abdul Halim, A.H.; Zainuddin, N. Carboxymethyl Cellulose Hydrogel from Biomass Waste of oil Palm Empty Fruit Bunch Using Calcium Chloride as Crosslinking Agent. Polymers 2021, 13, 4056.
  10. Omonije, O.O.; Egwim, E.C.; Kabiru, A.Y.; Olutoye, M.A. Sugarcane Bagasse as Carbon Source for the Production of Cellulase by Aspergillus Niger. BIOMED Nat. Appl. Sci. 2022, 2, 19–27.
  11. Raul, C.; Bharti, V.S.; Dar Jaffer, Y.; Lenka, S.; Krishna, G. Sugarcane Bagasse Biochar: Suitable Amendment for Inland Aquaculture Soils. Aquacult. Res. 2020, 52, 643–654.
  12. Biró, E.; Németh, A.S.; Sisak, C.; Feczkó, T.; Gyenis, J. Preparation of Chitosan Particles Suitable for Enzyme Immobilization. J. Biochem. Bioph. Methods 2008, 70, 1240–1246.
  13. Pandey, D.; Daverey, A.; Arunachalam, K. Biochar: Production, Properties and Emerging Role as a Support for Enzyme Immobilization. J. Cleaner Prod. 2020, 255, 120267.
  14. Zang, L.; Qiao, X.; Hu, L.; Yang, C.; Liu, Q.; Wei, C.; Qiu, J.; Mo, H.; Song, G.; Yang, J.; Liu, C. Preparation and Evaluation of Coal Fly Ash/Chitosan Composites as Magnetic Supports for Highly Efficient Cellulase Immobilization and Cellulose Bioconversion. Polymers 2018, 10, 523.
  15. Weng, Z.-H.; Nargotra, P.; Kuo, C.-H.; Liu, Y.-C. Immobilization of Recombinant Endoglucanase (CelA) from Clostridium thermocellum on Modified Regenerated Cellulose Membrane. Catalysts 2022, 12, 1356.
  16. Zhao, H.; Wang, L.; Bai, Y.; Li, Y.; Tang, T.; Liang, H.; Gao, D. Immobilized Enzyme with Sustainable Chestnut Biochar to Remediate Polycyclic Aromatic Hydrocarbons Contaminated Soils. Environ. Technol. 2024, 45, 2034–2044.
  17. Qamar, S.A.; Asgher, M.; Bilal, M. Immobilization of Alkaline Protease from Bacillus brevis Using Ca-Alginate Entrapment Strategy for Improved Catalytic Stability, Silver Recovery, and Dehairing Potentialities. Catal. Lett. 2020, 150, 3572–3583.
  18. Kurayama, F.; Mohammed Bahadur, N.; Furusawa, T.; Sato, M.; Suzuki, N. Facile Preparation of Aminosilane-alginate Hybrid Beads for Enzyme Immobilization: Kinetics and Equilibrium Studies. Int. J. Biol. Macromol. 2020, 150, 1203–1212.
  19. Saha, K.; Verma, P.; Sikder, J.; Chakraborty, S.; Curcio, S. Synthesis of Chitosan-cellulase Nanohybrid and Immobilization on Alginate Beads for Hydrolysis of Ionic Liquid Pretreated Sugarcane Bagasse. Renewable Energy 2019, 133, 66–76.
  20. Hawaz, E.; Tafesse, M.; Tesfaye, A.; Kiros, S.; Beyene, D.; Kebede, G.; Boekhout, T.; Groenwald, M.; Theelen, B.; Degefe, A.; et al. Optimization of Bioethanol Production from Sugarcane Molasses by the Response Surface Methodology Using Meyerozyma Caribbica Isolate MJTm3. Ann. Microbiol. 2023, 73, 2.
  21. Khan, M.R. Immobilized Enzymes: a Comprehensive Review. Bull. National Res. Centre 2021. 45, 1–13.
  22. Bié, J.; Sepodes, B.; Fernandes, P.C.B.; Ribeiro, M.H.L. Enzyme Immobilization and Co-Immobilization: Main Framework, Advances and Some Applications. Processes 2022, 10, 494.
  23. Sonkar, K.; Singh, D.P. Biochemical Characterization and Thermodynamic Study of Lipase from Psychrotolerant Pseudomonas Punonensis. Biocatal. Agric. Biotechnol. 2020, 28, 101686.
  24. Kirupa Sankar, M.; Ravikumar, R.; Naresh Kumar, M.; Sivakumar, U. Development of Co-Immobilized Tri-Enzyme Biocatalytic System for One-Pot Pretreatment of Four Different Perennial Lignocellulosic Biomass and Evaluation of Their Bioethanol Production Potential. Bioresour. Technol. 2018, 269, 227–236.
  25. Awad, G.E.A.; Wehaidy, H.R.; Abd El Aty, A.A.; Hassan, M.E. A Novel Alginate–CMC Gel Beads for Efficient Covalent Inulinase Immobilization. Colloid. Polym. Sci. 2017, 295, 495–506.
  26. Abou Alsoaud, M.M.; Taher, M.A.; Hamed, A.M.; Elnouby, M.S.; Omer, A.M. Reusable Kaolin Impregnated Aminated Chitosan Composite Beads for Efficient Removal of Congo Red Dye: Isotherms, Kinetics and Thermodynamics Studies. Sci. Rep. 2022, 12, 12972.
  27. Abraham, R.E.; Verma, M.L.; Barrow, C.J.; Puri, M. Suitability of Magnetic Nanoparticle Immobilised Cellulases in Enhancing Enzymatic Saccharification of Pretreated Hemp Biomass. Biotechnol. Biofuels 2014, 7, 1–12.
  28. Ladole, M.R.; Pokale, P.B.; Varude, V.R.; Belokar, P.G.; Pandit, A.B. One Pot Clarification and Debittering of Grapefruit Juice Using Co-Immobilized Enzymes@chitosanMNPs. Int. J. Biol. Macromol. 2021, 167, 1297–1307.