Uncovering the role of leucine 59 in Renilla luciferase stability and activity with error-prone PCR: Implications for protein engineering.

A new variant of Renilla luciferase, named Met C-SRLuc 8, was obtained from a random mutagenesis library and expressed in Escherichia coli BL21 (DE3) plys and purified. The results of the enzyme's binding affinity, kinetic stability, and bioinformatic studies demonstrated that leucine 59, located within the hot-spot foldon in the N-terminal domain of the protein, plays a significant role in the stability and activity of Renilla luciferase. These findings may facilitate the engineering of different variants of this enzyme to achieve thermally stable versions for various biotechnological applications.

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Copyright © 2023. Published by Elsevier Inc.

Déclaration de conflit d'intérêts

Declaration of interest statement The authors declare no conflicts of interest related to manuscript entitled " Uncovering the Role of Leucine 59 in Renilla Luciferase Stability and Activity with Error-Prone PCR: Implications for Protein Engineering". The research was conducted in an unbiased manner, and the authors were not influenced by any financial or personal relationships that could have affected the interpretation of the results. All relevant sources of funding for this study have been disclosed, and the authors have no financial relationships with any organizations that could be perceived as having biased the research reported in this manuscript. All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Zahra Fanaei Kahrani. The first draft of the manuscript was written by Zahra Fanaei Kahrani and edited by Rahman Emamzadeh and Mahboobeh Nazari. All authors read and approved the final manuscript. Sincerely, Rahman Emamzadeh (Associate professor), Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran. [e-mail].