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Título
Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-Acetylglycyl-Clavaminic acid is an intermediate of clavulanic acid biosynthesis
Autor
Facultad/Centro
Área de conocimiento
Título de la revista
Applied and Environmental Microbiology
Número de la revista
22
Cita Bibliográfica
Álvarez-Álvarez, R., Rodríguez-García, A., Martínez-Burgo, Y., Martín, J. F., & Liras, P. (2018). Transcriptional studies on a Streptomyces clavuligerus oppA2 deletion mutant: N-acetylglycyl-clavaminic acid is an intermediate of clavulanic acid biosynthesis. Applied and environmental microbiology, 84(22), e01701-18. https://doi.org/10.1128/AEM.01701-18
Editorial
American Society for Microbiology (ASM)
Fecha
2018-10-30
ISSN
0099-2240
Resumen
[EN] The oppA2 gene encodes an oligopeptide-binding protein similar to the periplasmic substrate-binding proteins of the ABC transport systems. However, oppA2 is an orphan gene, not included in an ABC operon. This gene is located in the clavulanic acid (CA) gene cluster of Streptomyces clavuligerus and is essential for CA production. A transcriptomic study of the oppA2-null mutant S. clavuligerus ΔoppA2::aac showed changes in the expression levels of 233 genes from those in the parental strain. These include genes for ABC transport systems, secreted proteins, peptidases, and proteases. Expression of the clavulanic acid, clavam, and cephamycin C biosynthesis gene clusters was not significantly affected in the oppA2 deletion mutant. The genes for holomycin biosynthesis were upregulated 2-fold on average, and the level of upregulation increased to 43-fold in a double mutant lacking oppA2 and the pSCL4 plasmid. Strains in which oppA2 was mutated secreted into the culture the compound N-acetylglycyl-clavaminic acid (AGCA), a putative intermediate of CA biosynthesis. A culture broth containing AGCA, or AGCA purified by liquid chromatography-mass spectrometry (LC-MS), was added to the cultures of various non-CA-producing mutants. Mutants blocked in the early steps of the pathway restored CA production, whereas mutants altered in late steps did not, establishing that AGCA is a late intermediate of the biosynthetic pathway, which is released from the cells when the oligopeptide-binding protein OppA2 is not available
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Palabras clave
Peer review
SI
ID proyecto
- info:eu-repo/grantAgreement/MINECO//BIO2013-34723
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