Design and application of artificial rare L-lysine codons in Corynebacterium glutamicum

Yang, Cuiping; Peng, Zehao; Lü, Yang; Du, Bowen; Guo, Chuanzhuang; Sui, Songsen; Wang, Jianbin; Li, Junlin; Wang, Junqing; Li, Nan

doi:10.3389/fbioe.2023.1194511

Cited by 2 publications

(3 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To increase the heterologous expression of ACOX, the T7 promoter that regulates ACOX was replaced using Parg. The arginine rare tRNA UCU Parg promoter serves as a natural promoter, which is unaffected by inducers and can regulate the heterologous expression of ACOX more efficiently . Subsequently, CYP was integrated into the same vector to catalyze the hydroxylation of trans-2-decenoic and decanoic acids (Figure A).…”

Section: Results and Discussionmentioning

confidence: 99%

“…The arginine rare tRNA UCU Parg promoter serves as a natural promoter, which is unaffected by inducers and can regulate the heterologous expression of ACOX more efficiently. 38 Subsequently, CYP was integrated into the same vector to catalyze the hydroxylation of trans-2-decenoic and decanoic acids (Figure 2A). The vector was coexpressed in the E. coli/SK system.…”

Section: One-mentioning

confidence: 99%

See 1 more Smart Citation

Biosynthesis of 10-Hydroxy-2-decenoic Acid through a One-Step Whole-Cell Catalysis

Fang,

Xu,

Yang

et al. 2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

10-Hydroxy-2-decenoic acid (10-HDA) is an important component of royal jelly, known for its antimicrobial, antiinflammatory, blood pressure-lowering, and antiradiation effects. Currently, 10-HDA biosynthesis is limited by the substrate selectivity of acyl-coenzyme A dehydrogenase, which restricts the technique to a two-step process. This study aimed to develop an efficient and simplified method for synthesizing 10-HDA. In this study, ACOX from Candida tropicalis 1798, which catalyzes 10hydroxydecanoyl coenzyme A desaturation for 10-HDA synthesis, was isolated and heterologously coexpressed with FadE, Macs, YdiI, and CYP in Escherichia coli/SK after knocking out FadB, FadJ, and FadR genes. The engineered E. coli/AKS strain achieved a 49.8% conversion of decanoic acid to 10-HDA. CYP expression was improved through ultraviolet mutagenesis and high-throughput screening, increased substrate conversion to 75.6%, and the synthesis of 10-HDA was increased to 0.628 g/L in 10 h. This is the highest conversion rate and product concentration achieved in the shortest time to date. This study provides a simple and efficient method for 10-HDA biosynthesis and offers an effective method for developing strains with high product yields.

show abstract

Section: Results and Discussionmentioning

confidence: 99%

Section: One-mentioning

confidence: 99%

Biosynthesis of 10-Hydroxy-2-decenoic Acid through a One-Step Whole-Cell Catalysis

Fang,

Xu,

Yang

et al. 2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

show abstract

“…In our previous research, we used eGFP as a screening marker in C. glutamicum [24]. However, this approach is not applicable for use with E. coli.…”

Section: Introductionmentioning

confidence: 99%

Screening l-Lysine-Overproducing Escherichia coli Using Artificial Rare Codons and a Rare Codon-Rich Marker

Liu,

Yang,

Yang

et al. 2023

Fermentation

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Design and application of artificial rare L-lysine codons in Corynebacterium glutamicum

Cited by 2 publications

References 35 publications

Biosynthesis of 10-Hydroxy-2-decenoic Acid through a One-Step Whole-Cell Catalysis

Biosynthesis of 10-Hydroxy-2-decenoic Acid through a One-Step Whole-Cell Catalysis

Screening l-Lysine-Overproducing Escherichia coli Using Artificial Rare Codons and a Rare Codon-Rich Marker

Contact Info

Product

Resources

About