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Microbiology and Biotechnology Letters

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Microbial Biotechnology  |  Protein Structure, Function, and Engineering

Microbiol. Biotechnol. Lett.

Received: March 27, 2022; Revised: April 15, 2022

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

Ho-Dong Lim , So-Young Han , Gi-Hye Park , Dae-Eun Cheong and Geun-Joong Kim *

Chonnam National University

Correspondence to :
Geun-Joong Kim, Department of Biological Sciences, Chonnam National University, 77, Youngbong-ro, Buk-gu, Gwangju 61186, Republic of Korea [61186]
Tel : 82-62-530-3403, Fax : 82-62-530-3409, E-mail : gjkim@chonnam.ac.kr

Abstract

Precisely reliable and quantitative reporters can provide phenotypes that are consistent with research goals in protein expression. We here developed an improved reporter mATglu III 5 by directed evolution using a versatile ?-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. mATglu III 5 was also analyzed to be fully functional in fusion state with oligomeric proteins, especially under non-induction conditions, thereby offering an alternative to conventional reporter.

Keywords: Versatile reporter, β-glucosidase, directed evolution, activity staining

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