Single-cell-based screening and engineering of D-amino acid amidohydrolases using artificial amidophenol substrates and microbial biosensors = 인공 아미도페놀 기질 및 바이오센서를 이용한 D-아미노산 아미도하이드롤라제의 단일세포기반 개량

Cited 1 time in scopus
Metadata Downloads
Title
Single-cell-based screening and engineering of D-amino acid amidohydrolases using artificial amidophenol substrates and microbial biosensors = 인공 아미도페놀 기질 및 바이오센서를 이용한 D-아미노산 아미도하이드롤라제의 단일세포기반 개량
Author(s)
S J Yeom; Kil Koang Kwon; Jung Ung An; Sung Hyun Park; Jin-Young Lee; Eugene RhaHyewon LeeHaseong KimDae Hee LeeSeung Goo Lee
Bibliographic Citation
Journal of Agricultural and Food Chemistry, vol. 70, no. 4, pp. 1203-1211
Publication Year
2022
Abstract
Enantiomerically pure d-amino acids are important intermediates as chiral building blocks for peptidomimetics and semisynthetic antibiotics. Here, a transcriptional factor-based screening strategy was used for the rapid screening of d-stereospecific amino acid amidase via an enzyme-specific amidophenol substrate. We used a d-threonine amidophenyl derivative to produce 2-aminophenol that serves as a putative enzyme indicator in the presence of d-threonine amidases. Comparative analyses of known bacterial species indicated that several Bacillus strains produce amidase and form putative indicators in culture media. The estimated amidase was cloned and subjected to rapid directed evolution through biosensor cells. Consequently, we characterized the F119A mutation that significantly improved the catalytic activity toward d-alanine, d-threonine, and d-glutamate. Its beneficial effects were confirmed by higher conversions and recurrent applications of the mutant enzyme, compared to the wild-type. This study showed that rapid directed evolution with biosensors coupled to designed substrates is useful to develop biocatalytic processes.
Keyword
AmidaseD-amino acidArtificial substrateBiosensorGenetic circuitHigh-throughput screening
ISSN
0021-8561
Publisher
Amer Chem Soc
DOI
http://dx.doi.org/10.1021/acs.jafc.1c05834
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.