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Directed Evolution of a β-Glucosidase for Improved Functions as a Reporter in Protein Expression

  • Lim, Ho-Dong (Department of Biological Sciences and Research Center of Ecomimetics, College of Natural Sciences, Chonnam National University) ;
  • Han, So-Young (Department of Biological Sciences and Research Center of Ecomimetics, College of Natural Sciences, Chonnam National University) ;
  • Park, Gi-Hye (Department of Biological Sciences and Research Center of Ecomimetics, College of Natural Sciences, Chonnam National University) ;
  • Cheong, Dae-Eun (Department of Biological Sciences and Research Center of Ecomimetics, College of Natural Sciences, Chonnam National University) ;
  • Kim, Geun-Joong (Department of Biological Sciences and Research Center of Ecomimetics, College of Natural Sciences, Chonnam National University)
  • Received : 2022.03.28
  • Accepted : 2022.04.18
  • Published : 2022.06.28

Abstract

Precisely reliable and quantitative reporters can provide phenotypes that are consistent with research goals in protein expression. Here, we developed an improved reporter mATglu III 5 by directed evolution using a versatile β-glucosidase ATglu derived from Agrobacterium tumefaciens. When expressed in hosts, a vector containing this mutant distinctly showed a colored or fluorescent phenotype, according to the supplemented substrate, without any inducer. Analysis of mATglu III 5 showed it to be fully functional in fusion state with oligomeric proteins, especially under non-induction conditions, thereby offering an alternative to conventional reporters.

Keywords

Acknowledgement

This work was supported by a grant (grant number: 20170305) from the Marine Biotechnology Program funded by the Ministry of Oceans and Fisheries, Korea. This work was also supported by a grant (grant number: 2021R1A2C1006734) of the Basic Science Research program of the National Research Foundation (NRF) funded by the Ministry of Education, Science and Technology of Korea (MEST), Republic of Korea.

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