Novel Mode Engineering for β-Alanine Production in Escherichia coli with the Guide of Adaptive Laboratory Evolution
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Plasmids
2.2. Media and Culture Conditions
2.3. Adaptive Laboratory Evolution
2.3.1. Continuous Adaptive Laboratory Evolution
2.3.2. Marginal Effect-Assisted Adaptive Laboratory Evolution
2.4. Measurement of β-Alanine and L-Aspartic Acid
3. Results
3.1. Evolution of PPC Deficient E. Coli Strains
3.2. Establishment of the GML by Whole Genome Sequencing of the Adapted Mutant Pool
3.3. Construction of the Cell Factory for the Biosynthesis of β-Alanine
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Strains | Relevant Characteristics | Source or Reference |
---|---|---|
B0016-082BB | B0016-080BB, panD:: CgpanD | Lab stock |
B0016-082B | B0016-082BB, aspA::FRT | This study |
B0016-090BB | B0016-082BB, ppc::FRT | This study |
B0016-090BBS1 | B0016-090BB, ALE mutant | This study |
B0016-090BBS2 | B0016-090BB, ALE mutant | This study |
B0016-090BBS3 | B0016-090BB, ALE mutant | This study |
B0016-090BBS4 | B0016-090BB, ALE mutant | This study |
B0016-090BBS5 | B0016-090BB, ALE mutant | This study |
B0016-100BB | B0016-090BB, recE::FRT, mhpF::FRT, ykgF::FRT, mhpB:: mhpB *, mhpD:: mhpD *, rcsA:: rcsA * | This study |
B0016-200BB | B0016-100BB, aspA::FRT | This study |
B0016-082B/pET24a-panD-AspDH | B0016-082B with pPL451-panD-AspDH plasmid | This study |
B0016-200BB/pET24a-panD-AspDH | B0016-200BB with pPL451-panD-AspDH plasmid | This study |
Genes | Mutations | Amino Acid Mutations |
---|---|---|
recE | 1737–1737 del a, 1739–1741 del | Frameshift mutation |
mhpB | A107C b | E36A d |
mhpD | T278C | I93T |
mhpF | T179C, G189C, 76–77 ins GC c, 116–117 del, 120–121 ins AA | Frameshift mutation |
ykgF | A19G, T27A,33–35 del, 37–38 ins T, 39–40 ins GT, C94G, | Frameshift mutation |
rcsA | T209A | L70H |
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Xu, J.; Zhou, L.; Yin, M.; Zhou, Z. Novel Mode Engineering for β-Alanine Production in Escherichia coli with the Guide of Adaptive Laboratory Evolution. Microorganisms 2021, 9, 600. https://doi.org/10.3390/microorganisms9030600
Xu J, Zhou L, Yin M, Zhou Z. Novel Mode Engineering for β-Alanine Production in Escherichia coli with the Guide of Adaptive Laboratory Evolution. Microorganisms. 2021; 9(3):600. https://doi.org/10.3390/microorganisms9030600
Chicago/Turabian StyleXu, Jian, Li Zhou, Meng Yin, and Zhemin Zhou. 2021. "Novel Mode Engineering for β-Alanine Production in Escherichia coli with the Guide of Adaptive Laboratory Evolution" Microorganisms 9, no. 3: 600. https://doi.org/10.3390/microorganisms9030600