Identification of a TetR family regulator and a polyketide synthase gene cluster involved in growth development and butenyl-spinosyn biosynthesis of Saccharopolyspora pogona.
Butenyl-spinosyn
CRISPR/Cas9 system
Metabolic engineering
Proteomics analysis
Saccharopolyspora pogona
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Feb 2021
Feb 2021
Historique:
received:
03
08
2020
accepted:
05
01
2021
revised:
04
12
2020
pubmed:
24
1
2021
medline:
15
5
2021
entrez:
23
1
2021
Statut:
ppublish
Résumé
Butenyl-spinosyn produced by Saccharopolyspora pogona exhibits strong insecticidal activity and broad pesticidal spectrum. However, its synthetic level was low in the wild-type strain. At present, important functional genes involved in butenyl-spinosyn biosynthesis remain unknown, which leads to difficulty in efficiently editing its genome to improve the butenyl-spinosyn yield. To accelerate the genetic modification of S. pogona, we conducted comparative proteomics analysis to screen differentially expressed proteins related to butenyl-spinosyn biosynthesis. A TetR family regulatory protein was selected from the 289 differentially expressed proteins, and its encoding gene (SP_1288) was successfully deleted by CRISPR/Cas9 system. We further deleted a 32-kb polyketide synthase gene cluster (cluster 28) to reduce the competition for precursors. Phenotypic analysis revealed that the deletion of the SP_1288 and cluster 28 resulted in a 3.10-fold increase and a 35.4% decrease in the butenyl-spinosyn levels compared with the wild-type strain, respectively. The deletion of cluster 28 affected the cell growth, glucose consumption, mycelium morphology, and sporulation by controlling the expression of ptsH, ptsI, amfC, and other genes related to sporulation, whereas SP_1288 did not. These findings confirmed not only that the CRISPR/Cas9 system can be applied to the S. pogona genome editing but also that SP_1288 and cluster 28 are closely related to the butenyl-spinosyn biosynthesis and growth development of S. pogona. The strategy reported here will be useful to reveal the regulatory mechanism of butenyl-spinosyn and improve antibiotic production in other actinomycetes. KEY POINTS: • SP_1288 deletion can significantly promote the butenyl-spinosyn biosynthesis. • Cluster 28 deletion showed pleiotropic effects on S. pogona. • SP_1288 and cluster 28 were deleted by CRISPR/Cas9 system in S. pogona.
Identifiants
pubmed: 33484320
doi: 10.1007/s00253-021-11105-4
pii: 10.1007/s00253-021-11105-4
doi:
Substances chimiques
Macrolides
0
Polyketide Synthases
79956-01-7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1519-1533Subventions
Organisme : National Natural Science Foundation of China
ID : 31770106
Organisme : National Natural Science Foundation of China
ID : 3107006
Organisme : National Basic Research Program (973) of China
ID : 2012CB722301
Organisme : National High Technology Research and Development program (863) of China
ID : 2011AA10A203
Organisme : Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province
ID : 20134486
Organisme : Key Research and Development Program of Human Province
ID : 2019NK2192
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