Publication: Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation
Loading...
Files
Date
2022-07-01
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
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.
Description
Keywords
Yaşam Bilimleri, Moleküler Biyoloji ve Genetik, Sitogenetik, Kimya, Fizikokimya, Polimer Karakterizasyonu, Diğer, Temel Bilimler, Life Sciences, Molecular Biology and Genetics, Cytogenetic, Chemistry, Physical Chemistry, Characterization of Polymers, Other, Natural Sciences, BİYOKİMYA VE MOLEKÜLER BİYOLOJİ, Moleküler Biyoloji ve Genetik, Yaşam Bilimleri (LIFE), KİMYA, UYGULAMALI, Temel Bilimler (SCI), POLİMER BİLİMİ, BIOCHEMISTRY & MOLECULAR BIOLOGY, MOLECULAR BIOLOGY & GENETICS, Life Sciences (LIFE), CHEMISTRY, APPLIED, CHEMISTRY, Natural Sciences (SCI), POLYMER SCIENCE, Biyokimya, Genetik ve Moleküler Biyoloji (çeşitli), Klinik Biyokimya, Kanser Araştırmaları, Moleküler Biyoloji, İlaç Keşfi, Yaşlanma, Genel Biyokimya, Genetik ve Moleküler Biyoloji, Biyokimya, Yapısal Biyoloji, Polimerler ve Plastikler, Kimya (çeşitli), Genel Kimya, Proses Kimyası ve Teknolojisi, Fizik Bilimleri, Biochemistry, Genetics and Molecular Biology (miscellaneous), Clinical Biochemistry, Cancer Research, Molecular Biology, Drug Discovery, Aging, General Biochemistry, Genetics and Molecular Biology, Biochemistry, Structural Biology, Polymers and Plastics, Chemistry (miscellaneous), General Chemistry, Process Chemistry and Technology, Physical Sciences, Computational analysis, Protein engineering, Enzyme immobilisation, Protein orientation, Protein secondary structure, HETEROFUNCTIONAL CARRIER, COVALENT IMMOBILIZATION, CANDIDA-RUGOSA, STABILITY, LIPASE, ORIENTATION, STRATEGIES, SUPPORTS, SITE, BENTONITE
Citation
Jaafar N. R., Jailani N., Rahman R. A., TOKSOY ÖNER E., Murad A. M. A., Illias R. M., "Protein surface engineering and interaction studies of maltogenic amylase towards improved enzyme immobilisation", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, cilt.213, ss.70-82, 2022