Screening l-Lysine-Overproducing Escherichia coli Using Artificial Rare Codons and a Rare Codon-Rich Marker Article Swipe
Hui Liu
,
Cuiping Yang
,
Lu Yang
,
Ruiming Wang
,
Piwu Li
,
Bowen Du
,
Nan Li
,
Junqing Wang
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.3390/fermentation9100899
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.3390/fermentation9100899
l-Lysine, an essential amino acid for humans and mammals, is widely used in the food, feed, medicine, and cosmetics industries. In this study, a lysine over-producing Escherichia coli mutant was isolated using a fluorescence-based screen and an E. coli strain lacking five of the six L-lysine tRNA-UUU genes. Firstly, an l-lysine codon-rich protein was fused with a green fluorescent protein (all AAG codons were replaced with AAA), yielding a rare codon-rich screening marker positively correlated with l-lysine content. After association and room temperature plasma (ARTP) mutagenesis and induced fluorescent protein expression culture, mutant strains with strong fluorescence were sorted using flow cytometry. The fermentation performance of the high-yielding l-lysine strains were evaluated, which resulted in 16 of the 29 mutant strains showing increased L-lysine yields compared with those of the wild-type strains and a screening efficiency of up to 55.2%. Following a 48 h fermentation, the production of l-lysine (14.8 g/L) and biomass by E. coli QD01ΔtRNA L2 were 12.1 and 4.5% higher than those of the wild-type strain. The screening strategy for high-yielding strains based on the artificial rare cryptosystem established in this study will provide an efficient, accurate, and simple method for screening other amino-acid-producing microorganisms.
Related Topics
Concepts
Lysine
Escherichia coli
Biology
Mutant
Amino acid
Biochemistry
Green fluorescent protein
Mutagenesis
Transfer RNA
Gene
RNA
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3390/fermentation9100899
- https://www.mdpi.com/2311-5637/9/10/899/pdf?version=1696906210
- OA Status
- gold
- Cited By
- 4
- References
- 38
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4387494204
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4387494204Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.3390/fermentation9100899Digital Object Identifier
- Title
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Screening l-Lysine-Overproducing Escherichia coli Using Artificial Rare Codons and a Rare Codon-Rich MarkerWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2023Year of publication
- Publication date
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2023-10-10Full publication date if available
- Authors
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Hui Liu, Cuiping Yang, Lu Yang, Ruiming Wang, Piwu Li, Bowen Du, Nan Li, Junqing WangList of authors in order
- Landing page
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https://doi.org/10.3390/fermentation9100899Publisher landing page
- PDF URL
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https://www.mdpi.com/2311-5637/9/10/899/pdf?version=1696906210Direct link to full text PDF
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YesWhether a free full text is available
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goldOpen access status per OpenAlex
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https://www.mdpi.com/2311-5637/9/10/899/pdf?version=1696906210Direct OA link when available
- Concepts
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Lysine, Escherichia coli, Biology, Mutant, Amino acid, Biochemistry, Green fluorescent protein, Mutagenesis, Transfer RNA, Gene, RNATop concepts (fields/topics) attached by OpenAlex
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4Total citation count in OpenAlex
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2025: 2, 2024: 2Per-year citation counts (last 5 years)
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10Other works algorithmically related by OpenAlex
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