Effect of exogenous lysine on the expression of early cephamycin C biosynthetic genes and antibiotic production in Nocardia lactamdurans MA4213

2001 ◽  
Vol 56 (5-6) ◽  
pp. 670-675 ◽  
Author(s):  
A. L. Leitão ◽  
F. J. Enguita ◽  
J. F. Martín ◽  
J. F. Santos Oliveira
Keyword(s):  
Antibiotic Production ◽  
Biosynthetic Genes ◽  
Cephamycin C ◽  
Nocardia Lactamdurans

scholarly journals Inducing Effect of Diamines on Transcription of the Cephamycin C Genes from the lat and pcbAB Promoters inNocardia lactamdurans

1999 ◽  
Vol 181 (8) ◽  
pp. 2379-2384 ◽  
Author(s):  
Ana Lucia Leitão ◽  
Francisco J. Enguita ◽  
Juan Luis De La Fuente ◽  
Paloma Liras ◽  
Juan F. Martin
Keyword(s):  
Cell Growth ◽  
Antibiotic Production ◽  
Protein A ◽  
Transcriptional Level ◽  
Intracellular Accumulation ◽  
Low Resolution ◽  
Cephamycin C ◽  
Nocardia Lactamdurans ◽  
Shake Flasks ◽  
Acid Precursor

ABSTRACT The diamines putrescine, cadaverine, and diaminopropane stimulate cephamycin biosynthesis in Nocardia lactamdurans, in shake flasks and fermentors, without altering cell growth. Intracellular levels of the P7 protein (a component of the methoxylation system involved in cephamycin biosynthesis) were increased by diaminopropane, as shown by immunoblotting studies. Lysine-6-aminotransferase and piperideine-6-carboxylate dehydrogenase activities involved in biosynthesis of the α-aminoadipic acid precursor were also greatly stimulated. The diamine stimulatory effect is exerted at the transcriptional level, as shown by low-resolution S1 protection studies. The transcript corresponding to the pcbAB gene and to a lesser extent also the lat transcript were significantly increased in diaminopropane-supplemented cultures, whereas transcription from the cefD promoter was not affected. Coupling of the lat and pcbABpromoters to the reporter xylE gene showed that expression from the lat and pcbAB promoters was increased by addition of diaminopropane in Streptomyces lividans. Intracellular accumulation of diamines in Nocardia may be a signal to trigger antibiotic production.

scholarly journals The Streptomyces filipinensis Gamma-Butyrolactone System Reveals Novel Clues for Understanding the Control of Secondary Metabolism

2020 ◽  
Vol 86 (18) ◽  
Author(s):  
Eva G. Barreales ◽  
Tamara D. Payero ◽  
Ester Jambrina ◽  
Jesús F. Aparicio
Keyword(s):  
Quorum Sensing ◽  
Secondary Metabolism ◽  
Target Genes ◽  
Antibiotic Production ◽  
The Other ◽  
Antimycin A ◽  
Dependent Manner ◽  
Biosynthetic Genes ◽  
Content Type ◽  
Communication Signals

ABSTRACT Streptomyces γ-butyrolactones (GBLs) are quorum sensing communication signals triggering antibiotic production. The GBL system of Streptomyces filipinensis, the producer of the antifungal agent filipin, has been investigated. Inactivation of sfbR (for S. filipinensis γ-butyrolactone receptor), a GBL receptor, resulted in a strong decrease in production of filipin, and deletion of sfbR2, a pseudo-receptor, boosted it, in agreement with lower and higher levels of transcription of filipin biosynthetic genes, respectively. It is noteworthy that none of the mutations affected growth or morphological development. While no ARE (autoregulatory element)-like sequences were found in the promoters of filipin genes, suggesting indirect control of production, five ARE sequences were found in five genes of the GBL cluster, whose transcription has been shown to be controlled by both S. filipinensis SfbR and SfbR2. In vitro binding of recombinant SfbR and SfbR2 to such sequences indicated that such control is direct. Transcription start points were identified by 5′ rapid amplification of cDNA ends, and precise binding regions were investigated by the use of DNase I protection studies. Binding of both regulators took place in the promoter of target genes and at the same sites. Information content analysis of protected sequences in target promoters yielded an 18-nucleotide consensus ARE sequence. Quantitative transcriptional analyses revealed that both regulators are self-regulated and that each represses the transcription of the other as well as that of the remaining target genes. Unlike other GBL receptor homologues, SfbR activates its own transcription whereas SfbR2 has a canonical autorepression profile. Additionally, SfbR2 was found here to bind the antifungal antimycin A as a way to modulate its DNA-binding activity. IMPORTANCE Streptomyces GBLs are important signaling molecules that trigger antibiotic production in a quorum sensing-dependent manner. We have characterized the GBL system from S. filipinensis, finding that two key players of this system, the GBL receptor and the pseudo-receptor, each counteracts the transcription of the other for the modulation of filipin production and that such control over antifungal production involves an indirect effect on the transcription of filipin biosynthetic genes. Additionally, the two regulators bind the same sites, are self-regulated, and repress the transcription of three other genes of the GBL cluster, including that encoding the GBL synthase. In contrast to all the GBL receptors known, SfbR activates its own synthesis. Moreover, the pseudo-receptor was identified as the receptor of antimycin A, thus extending the range of examples supporting the idea of signaling effects of antibiotics in Streptomyces. The intricate regulatory network depicted here should provide important clues for understanding the regulatory mechanism governing secondary metabolism.

Δ-1-Piperideine-6-carboxylate dehydrogenase, a new enzyme that forms α-aminoadipate in Streptomyces clavuligerus and other cephamycin C-producing actinomycetes

1997 ◽  
Vol 327 (1) ◽  
pp. 59-64 ◽  
Author(s):  
Juan L. de la FUENTE ◽  
Angel RUMBERO ◽  
Juan F. MARTÍN ◽  
Paloma LIRAS
Keyword(s):  
Dehydrogenase Activity ◽  
Maximal Activity ◽  
Streptomyces Clavuligerus ◽  
Electron Acceptors ◽  
Native Protein ◽  
Cephamycin C ◽  
Second Stage ◽  
Streptomyces Cattleya ◽  
Nocardia Lactamdurans ◽  
Blue Sepharose

Δ-1-Piperideine-6-carboxylate (P6C) dehydrogenase activity, which catalyses the conversion of P6C into α-aminoadipic acid, has been studied in the cephamycin C producer Streptomyces clavuligerus by both spectrophotometric and radiometric assays. The enzyme has been purified 124-fold to electrophoretic homogeneity with a 26% yield. The native protein is a monomer of 56.2 kDa that efficiently uses P6C (apparent Km 14 μM) and NAD+ (apparent Km 115 μM), but not NADP+ or other electron acceptors, as substrates. The enzyme activity was inhibited (by 66%) by its end product NADH at 0.1 mM concentration. It did not show activity towards pyrroline-5-carboxylate and was separated by Blue-Sepharose chromatography from pyrroline-5-carboxylate dehydrogenase, an enzyme involved in the catabolism of proline. P6C dehydrogenase reached maximal activity later than other early enzymes of the cephamycin pathway. The P6C dehydrogenase activity was decreased in ammonium (40 mM)-supplemented cultures, as was that of lysine 6-aminotransferase. P6C dehydrogenase activity was also found in other cephamycin C producers (Streptomyces cattleya and Nocardia lactamdurans) but not in actinomycetes that do not produce β-lactams, suggesting that it is an enzyme specific for cephamycin biosynthesis, involved in the second stage of the two-step conversion of lysine to α-aminoadipic acid.

Overexpression of the lat gene in Nocardia lactamdurans from strong heterologous promoters results in very high levels of lysine-6-aminotransferase and up to two-fold increase in cephamycin C production

2000 ◽  
Vol 53 (3) ◽  
pp. 282-288 ◽  
Author(s):  
V. K. Chary ◽  
J. L. de la Fuente ◽  
A. L. Leitão ◽  
P. Liras ◽  
J. F. Martín
Keyword(s):  
Fold Increase ◽  
Cephamycin C ◽  
Nocardia Lactamdurans ◽  
Very High

scholarly journals The autoregulator receptor homologue AvaR3 plays a regulatory role in antibiotic production, mycelial aggregation and colony development of Streptomyces avermitilis

Microbiology ◽  
2011 ◽  
Vol 157 (8) ◽  
pp. 2266-2275 ◽  
Author(s):  
Kiyoko T. Miyamoto ◽  
Shigeru Kitani ◽  
Mamoru Komatsu ◽  
Haruo Ikeda ◽  
Takuya Nihira
Keyword(s):  
Antibiotic Production ◽  
Genomic Analysis ◽  
Morphological Differentiation ◽  
Gene Clusters ◽  
Streptomyces Avermitilis ◽  
Colony Development ◽  
Amino Acid Residues ◽  
Biosynthetic Genes ◽  
Morphological Defects ◽  
Autoregulator Receptor

The γ-butyrolactone autoregulator receptor has been shown to control secondary metabolism and/or morphological differentiation across many Streptomyces species. Streptomyces avermitilis produces an important anthelmintic agent (avermectin) and two further polyketide antibiotics, filipin and oligomycin. Genomic analysis of S. avermitilis revealed that this micro-organism has the clustered putative autoregulator receptor genes distant from the antibiotic biosynthetic gene clusters. Here, we describe the characterization of avaR3, one of the clustered receptor genes, which encodes a protein containing an extra stretch of amino acid residues that has not been found in the family of autoregulator receptors. Disruption of avaR3 resulted in markedly decreased production of avermectins, with delayed expression of avermectin biosynthetic genes, suggesting that AvaR3 positively controls the avermectin biosynthetic genes. Moreover, the disruption caused increased production of filipin without any changes in the transcriptional profile of the filipin biosynthetic genes, suggesting that filipin production is indirectly controlled by AvaR3. The avaR3 disruptant displayed fragmented growth in liquid culture and conditional morphological defects on solid medium. These findings demonstrated that AvaR3 acts as a global regulator that controls antibiotic production and cell morphology.

Identification of transcriptional activators for thienamycin and cephamycin C biosynthetic genes within the thienamycin gene cluster fromStreptomyces cattleya

2008 ◽  
Vol 69 (3) ◽  
pp. 633-645 ◽  
Author(s):  
Miriam Rodríguez ◽  
Luz Elena Núñez ◽  
Alfredo F. Braña ◽  
Carmen Méndez ◽  
José A. Salas ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Transcriptional Activators ◽  
Biosynthetic Genes ◽  
Cephamycin C
Sign in / Sign up

Export Citation Format

Share Document