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Enhancement of solubility of recombinant alcohol dehydrogenase from Rhodococcus ruber using predictive tool

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Abstract

Solubility is one of key factors influencing the heterologous production of recombinant proteins in biotechnology. Among many aggregation-prone proteins, alcohol dehydrogenase (ADH-A) from Rhodococcus ruber (in this work abbreviated RrADH) shows a great potential in processes involved in the biotransformation of natural compounds. As ADH-A is a potentially high value asset in industrial biotransformation processes, improvement of its solubility would be of major commercial benefit. Predictive tools and in silico analysis provide a fast means for improving protein properties, for selecting appropriate changes, and ultimately for saving costs. We have therefore focused on enhancement of the solubility of RrADH using an online accesible predictive tool Aggrescan 3D 2.0. Selected mutations were introduced into the protein amino acid sequence by using site-directed PCR. This led to a 17% increase in the protein solubility of RrADHmut1 and a 98% increase for RrADHmut2. Moreover, the basic kinetics of the enzyme reaction were positively affected, further optimizing the overall performance of the production process.

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Acknowledgements

We thank Dr. Theresa Jones for linguistic corrections of the manuscript.

Funding

This study was funded by APVV-15–0466—Preparation of natural flavors by biotransformation using the comprehensive analytical methods and APVV-17–0333—Research and development of effective processes for the preparation of vanillin and other natural flavors using the oxidative and protective effect of recombinant catalase and peroxidase. The work was also supported by the ERDF projects: Smartfarm (ITMS2014: 313011W112) and LISPER (ITMS2014: 313011V446).

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Authors and Affiliations

  1. Faculty of Natural Sciences, Comenius University in Bratislava, Ilkovičova 6, 811 04, Bratislava, Karlova Ves, Slovak Republic

    Andrej Minich, Zdenko Levarski & Stanislav Stuchlík

  2. Science Park, Comenius University in Bratislava, Ilkovičova 8, Bratislava, 811 04, Karlova Ves, Slovak Republic

    Júlia Šarkanová, Zdenko Levarski & Stanislav Stuchlík

Authors
  1. Andrej Minich
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  2. Júlia Šarkanová
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  3. Zdenko Levarski
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  4. Stanislav Stuchlík
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by AM and JŠ. The first draft of the manuscript was written by AM, JŠ and ZL. The draft of this manuscript was reviewed and edited by AM and ZL. Data curation, reviewing and editing of this manuscript was performed by AM, ZL and SS. Project was administrated and funding aqusistion provided by SS. All authors read and approved the final manuscript.

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Correspondence to Stanislav Stuchlík.

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Minich, A., Šarkanová, J., Levarski, Z. et al. Enhancement of solubility of recombinant alcohol dehydrogenase from Rhodococcus ruber using predictive tool. World J Microbiol Biotechnol 38, 214 (2022). https://doi.org/10.1007/s11274-022-03403-4

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