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Design of a dual-responding genetic circuit for high-throughput identification of L-threonine-overproducing Escherichia coli.

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Title: Design of a dual-responding genetic circuit for high-throughput identification of L-threonine-overproducing Escherichia coli.
Authors: Su, Buli1 (AUTHOR) bolysu@hotmail.com, Lai, Peixuan1 (AUTHOR) 522019158@qq.com, Deng, Ming-Rong1 (AUTHOR) dengmr@gdim.cn, Zhu, Honghui1 (AUTHOR) zhuhh_gdim@163.com
Source: Bioresource Technology. Mar2024, Vol. 395, pN.PAG-N.PAG. 1p.
Subjects: Threonine, Escherichia coli, Metabolic regulation, High throughput screening (Drug development), Animal feeds, Biosensors
Abstract: [Display omitted] • The inducer-like effect of L-threonine was firstly demonstrated. • A dual-responding genetic circuit was developed as L-threonine biosensor. • The developed HTS platform could identify mutants from a large-scale RBS library. • The HTS platform was applied in direction evolution of the key enzyme thrA. • L-threonine production increased 7-fold through directed evolution of the key enzyme. L-threonine is a crucial amino acid that is extensively employed in the realms of food, animal feed and pharmaceuticals. Unfortunately, the lack of an appropriate biosensor has hindered the establishment of a robust high-throughput screening (HTS) system for the identification of the desired strains from random mutants. In this study, a dual-responding genetic circuit that capitalizes on the L-threonine inducer-like effect, the L-threonine riboswitch, and a signal amplification system was designed for the purpose of screening L-threonine overproducers. This platform effectively enhanced the performance of the enzyme and facilitated the identification of high L-threonine-producing strains from a random mutant library. Consequently, pathway optimization and directed evolution of the key enzyme enhanced L-threonine production by 4 and 7-fold, respectively. These results demonstrate the potential of biosensor design for dynamic metabolite detection and offer a promising tool for HTS and metabolic regulation for the development of L-threonine-hyperproducing strains. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
Description
Abstract:[Display omitted] • The inducer-like effect of L-threonine was firstly demonstrated. • A dual-responding genetic circuit was developed as L-threonine biosensor. • The developed HTS platform could identify mutants from a large-scale RBS library. • The HTS platform was applied in direction evolution of the key enzyme thrA. • L-threonine production increased 7-fold through directed evolution of the key enzyme. L-threonine is a crucial amino acid that is extensively employed in the realms of food, animal feed and pharmaceuticals. Unfortunately, the lack of an appropriate biosensor has hindered the establishment of a robust high-throughput screening (HTS) system for the identification of the desired strains from random mutants. In this study, a dual-responding genetic circuit that capitalizes on the L-threonine inducer-like effect, the L-threonine riboswitch, and a signal amplification system was designed for the purpose of screening L-threonine overproducers. This platform effectively enhanced the performance of the enzyme and facilitated the identification of high L-threonine-producing strains from a random mutant library. Consequently, pathway optimization and directed evolution of the key enzyme enhanced L-threonine production by 4 and 7-fold, respectively. These results demonstrate the potential of biosensor design for dynamic metabolite detection and offer a promising tool for HTS and metabolic regulation for the development of L-threonine-hyperproducing strains. [ABSTRACT FROM AUTHOR]
ISSN:09608524
DOI:10.1016/j.biortech.2024.130407