Multiplex gene knockout raises Ala-Gln production by Escherichia coli expressing amino acid ester acyltransferase.
CRISPR-Cas9
Gene knockout
Large-scale Ala-Gln production
α-Amino acid ester acyltransferase
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Jun 2023
Jun 2023
Historique:
received:
25
10
2022
accepted:
19
04
2023
revised:
14
04
2023
medline:
15
5
2023
pubmed:
5
5
2023
entrez:
5
5
2023
Statut:
ppublish
Résumé
L-Alanyl-L-Glutamine (Ala-Gln) is a common parenteral nutritional supplement. In our previous study, the recombinant whole-cell catalyst Escherichia coli BL21(DE3) overexpressing α-amino acid ester acyltransferase (BPA) to produce Ala-Gln has high activity and has been applied to large-scale production experiments. However, the degradation of Ala-Gln is detected under prolonged incubation, and endogenous broad-spectrum dipeptidase may be the primary cause. In this study, a CRISPR-Cas9 method was used to target pepA, pepB, pepD, pepN, dpp, and dtp to knock out one or more target genes. The deletion combination was optimized, and a triple knockout strain BL21(DE3)-ΔpepADN was constructed. The degradation performance of the knockout chassis was measured, and the results showed that the degradation rate of Ala-Gln was alleviated by 48% compared with the control. On this basis, B
Identifiants
pubmed: 37145161
doi: 10.1007/s00253-023-12550-z
pii: 10.1007/s00253-023-12550-z
pmc: PMC10161157
doi:
Substances chimiques
Amino Acids
0
alanylglutamine
U5JDO2770Z
Dipeptidases
EC 3.4.13.-
Acyltransferases
EC 2.3.-
Dipeptides
0
Glutamine
0RH81L854J
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3523-3533Informations de copyright
© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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