Automated optimisation of the solubility of a hyper-stable α-amylase - presented by Montader Ali

Automated optimisation of the solubility of a hyper-stable α-amylase

Montader Ali

Montader Ali
Open Biology

Associated Open Biology article

M. Ali et al. (2024) Automated optimization of the solubility of a hyper-stable α-amylase. Open Biology
Article of record
Automated optimisation of the solubility of a hyper-stable α-amylase
Montader Ali
Montader Ali
University of Cambridge

Most successes of computational protein engineering to date have focused on enhancing one biophysical trait, while multi-trait optimisation remains a challenge. Different biophysical properties are often conflicting, as mutations that improve one tend to worsen the others. In this study, we explored the potential of an automated computational design strategy, called CamSol Combination, to optimise solubility and stability of enzymes without affecting their activity. Specifically, we focus on B. licheniformis α-amylase (BLA), a hyper-stable enzyme that finds diverse application in industry and biotechnology. We validate the computational predictions by producing 10 BLA variants, including the WT and 3 designed models harbouring between 6 and 8 mutations each. Our results show that all 3 models have substantially improved relative solubility over the WT, unaffected catalytic rate, and retained hyper-stability, supporting the algorithm's capacity to optimise enzymes. High stability and solubility embody enzymes with superior resilience to chemical and physical stresses, enhance manufacturability, and allow for high concentration formulations characterised by extended shelf-lives. This ability to readily optimise solubility and stability of enzymes will enable the rapid and reliable generation of highly robust and versatile reagents, poised to contribute to advancements in diverse scientific and industrial domains.

References
  • 1.
    A. Rosace et al. (2023) Automated optimisation of solubility and conformational stability of antibodies and proteins. Nature Communications
  • 2.
    P. Sormanni et al. (2014) The CamSol Method of Rational Design of Protein Mutants with Enhanced Solubility. Journal of Molecular Biology
  • 3.
    J. Schymkowitz et al. (2005) The FoldX web server: an online force field. Nucleic Acids Research
  • 4.
    M. Ali et al. (2024) Automated optimization of the solubility of a hyper-stable α-amylase. Open Biology
  • 5.
    A. Broom et al. (2017) Computational tools help improve protein stability but with a solubility tradeoff. Journal of Biological Chemistry
Grants
    Royal SocietyURF\R1\201461
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Cite as
M. Ali (2024, June 3), Automated optimisation of the solubility of a hyper-stable α-amylase
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Listed seminar This seminar is open to all
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Video length 16:21
Disclaimer The views expressed in this seminar are those of the speaker and not necessarily those of the journal