Recombineering using RecET-like recombinases from Xenorhabdus and its application in mining of natural products.
Changshamycin
Natural product
RecET-like recombinase
Recombineering
Xenorhabdus
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Dec 2022
Dec 2022
Historique:
received:
18
08
2022
accepted:
23
10
2022
revised:
14
10
2022
pubmed:
4
11
2022
medline:
19
11
2022
entrez:
3
11
2022
Statut:
ppublish
Résumé
Xenorhabdus can produce a large number of secondary metabolites with insecticidal, bacteriostatic, and antitumor activities. Efficient gene editing tools will undoubtedly facilitate the functional genomics research and bioprospecting in Xenorhabdus. In this study, BlastP analysis using the amino acid sequences of Redαβ or RecET recombinases as queries resulted in the identification of an operon (XBJ1_operon 0213) containing RecET-like recombinases encoding genes from the genome of Xenorhabdus bovienii strain SS-2004. Three proteins encoded by this operon was indispensable for full activity of recombineering, namely XBJ1-1173 (RecE-like protein), XBJ1-1172 (RecT-like protein), and XBJ1-1171 (single-strand annealing protein). Using this newly developed recombineering system, a gene cluster responsible for biosynthesis of a novel secondary metabolite (Min16) was identified from X. stockiae HN_xs01 strain. Min16 which exhibited antibacterial and cytotoxic activities was determined to be a cyclopeptide composed of Acyl-Phe-Thr-Phe-Pro-Pro-Leu-Val by using high-resolution mass spectrometry and nuclear magnetic resonance analysis, and was designated as changshamycin. This host-specific recombineering system was proven to be effective for gene editing in Xenorhabdus, allowing for efficient discovery of novel natural products with attractive bioactivities. KEY POINTS: • Screening and identification of efficient gene editing tools from Xenorhabdus • Optimization of the Xenorhabdus electroporation parameters • Discovery of a novel cyclopeptide compound with multiple biological activities.
Identifiants
pubmed: 36326838
doi: 10.1007/s00253-022-12258-6
pii: 10.1007/s00253-022-12258-6
doi:
Substances chimiques
Recombinases
0
Biological Products
0
Peptides, Cyclic
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7857-7866Subventions
Organisme : National Natural Science Foundation of China
ID : 32070090
Organisme : Research Foundation of Education Bureau of Hunan Province
ID : 19K053
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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