Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation
A combined strategy of computational, protein engineering and cross-linked enzyme aggregates (CLEAs) approaches was performed on Bacillus lehensis G1 maltogenic amylase (Mag1) to investigate the preferred amino acids and orientation of the cross-linker in constructing stable and efficient biocatalys...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Published: |
Elsevier B.V.
2022
|
| Subjects: |
| _version_ | 1796866981101568000 |
|---|---|
| author | Jaafar, Nardiah Rizwana Jailani, Nashriq A. Rahman, Roshanida Öner, Ebru Toksoy Abdul Murad, Abdul Munir Md. Illias, Rosli |
| author_facet | Jaafar, Nardiah Rizwana Jailani, Nashriq A. Rahman, Roshanida Öner, Ebru Toksoy Abdul Murad, Abdul Munir Md. Illias, Rosli |
| author_sort | Jaafar, Nardiah Rizwana |
| collection | ePrints |
| description | A combined strategy of computational, protein engineering and cross-linked enzyme aggregates (CLEAs) approaches was performed on Bacillus lehensis G1 maltogenic amylase (Mag1) to investigate the preferred amino acids and orientation of the cross-linker in constructing stable and efficient biocatalyst. From the computational analysis, Mag1 exhibited the highest binding affinity towards chitosan (−7.5 kcal/mol) and favours having interactions with aspartic acid whereas glutaraldehyde was the least favoured (−3.4 kcal/mol) and has preferences for lysine. A total of eight Mag1 variants were constructed with either Asp or Lys substitutions on different secondary structures surface. Mutant Mag1-mDh exhibited the highest recovery activity (82.3%) in comparison to other Mag1 variants. Mutants-CLEAs exhibited higher thermal stability (20–30% activity) at 80 °C whilst Mag1-CLEAs could only retain 9% of activity at the same temperature. Reusability analysis revealed that mutants-CLEAs can be recovered up to 8 cycles whereas Mag1-CLEAs activity could only be retained for up to 6 cycles. Thus, it is evident that amino acids on the enzyme's surface play a crucial role in the construction of highly stable, efficient and recyclable CLEAs. This demonstrates the necessity to determine the preferential amino acid by the cross-linkers in advance to facilitate CLEAs immobilisation for designing efficient biocatalysts. |
| first_indexed | 2024-03-05T21:20:19Z |
| format | Article |
| id | utm.eprints-100999 |
| institution | Universiti Teknologi Malaysia - ePrints |
| last_indexed | 2024-03-05T21:20:19Z |
| publishDate | 2022 |
| publisher | Elsevier B.V. |
| record_format | dspace |
| spelling | utm.eprints-1009992023-05-18T06:13:59Z http://eprints.utm.my/100999/ Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation Jaafar, Nardiah Rizwana Jailani, Nashriq A. Rahman, Roshanida Öner, Ebru Toksoy Abdul Murad, Abdul Munir Md. Illias, Rosli TP Chemical technology A combined strategy of computational, protein engineering and cross-linked enzyme aggregates (CLEAs) approaches was performed on Bacillus lehensis G1 maltogenic amylase (Mag1) to investigate the preferred amino acids and orientation of the cross-linker in constructing stable and efficient biocatalyst. From the computational analysis, Mag1 exhibited the highest binding affinity towards chitosan (−7.5 kcal/mol) and favours having interactions with aspartic acid whereas glutaraldehyde was the least favoured (−3.4 kcal/mol) and has preferences for lysine. A total of eight Mag1 variants were constructed with either Asp or Lys substitutions on different secondary structures surface. Mutant Mag1-mDh exhibited the highest recovery activity (82.3%) in comparison to other Mag1 variants. Mutants-CLEAs exhibited higher thermal stability (20–30% activity) at 80 °C whilst Mag1-CLEAs could only retain 9% of activity at the same temperature. Reusability analysis revealed that mutants-CLEAs can be recovered up to 8 cycles whereas Mag1-CLEAs activity could only be retained for up to 6 cycles. Thus, it is evident that amino acids on the enzyme's surface play a crucial role in the construction of highly stable, efficient and recyclable CLEAs. This demonstrates the necessity to determine the preferential amino acid by the cross-linkers in advance to facilitate CLEAs immobilisation for designing efficient biocatalysts. Elsevier B.V. 2022 Article PeerReviewed Jaafar, Nardiah Rizwana and Jailani, Nashriq and A. Rahman, Roshanida and Öner, Ebru Toksoy and Abdul Murad, Abdul Munir and Md. Illias, Rosli (2022) Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation. International Journal of Biological Macromolecules, 213 (-). pp. 70-82. ISSN 0141-8130 http://dx.doi.org/10.1016/j.ijbiomac.2022.05.169 DOI: 10.1016/j.ijbiomac.2022.05.169 |
| spellingShingle | TP Chemical technology Jaafar, Nardiah Rizwana Jailani, Nashriq A. Rahman, Roshanida Öner, Ebru Toksoy Abdul Murad, Abdul Munir Md. Illias, Rosli Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| title | Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| title_full | Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| title_fullStr | Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| title_full_unstemmed | Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| title_short | Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| title_sort | protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation |
| topic | TP Chemical technology |
| work_keys_str_mv | AT jaafarnardiahrizwana proteinsurfaceengineeringandinteractionstudiesofmaltogenicamylasetowardsimprovedenzymeimmobilisation AT jailaninashriq proteinsurfaceengineeringandinteractionstudiesofmaltogenicamylasetowardsimprovedenzymeimmobilisation AT arahmanroshanida proteinsurfaceengineeringandinteractionstudiesofmaltogenicamylasetowardsimprovedenzymeimmobilisation AT onerebrutoksoy proteinsurfaceengineeringandinteractionstudiesofmaltogenicamylasetowardsimprovedenzymeimmobilisation AT abdulmuradabdulmunir proteinsurfaceengineeringandinteractionstudiesofmaltogenicamylasetowardsimprovedenzymeimmobilisation AT mdilliasrosli proteinsurfaceengineeringandinteractionstudiesofmaltogenicamylasetowardsimprovedenzymeimmobilisation |