Transcriptome Analysis of an Antibiotic Downregulator Mutant and Synergistic Actinorhodin Stimulation via Disruption of a Precursor Flux Regulator in
            <i>Streptomyces coelicolor</i>

Kim, Seon-Hye; Lee, Hanna; Kim, Hye-Jin; Kim, Eung-Soo

doi:10.1128/aem.02346-10

Cited by 27 publications

(19 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3, real-time PCR analyses demonstrated that the levels of actII-orf4 transcripts were higher in the ⌬wblA mutant during the time course than in S. coelicolor M145 and the ⌬adpA mutant. It is consistent that the ⌬wblA mutant produced the highest level of actinorhodin and the ⌬adpA mutant did not produce actinorhodin at all (15,29,30). The levels of adpA transcripts from S. coelicolor M145 were similar to those in the ⌬wblA mutant, suggesting that the expression of adpA was not influenced by the wblA gene, and suggesting that the transcription of adpA occurs upstream of wblA transcription.…”

supporting

confidence: 59%

“…Based on these results, we wondered whether AdpA SC regulates wblA gene expression as an activator or repressor. To test this, transcription analysis was conducted with strains of S. coelicolor M145, the S. coelicolor ⌬wblA mutant (26,29), and the S. coelicolor ⌬adpA mutant (19). It was previously reported that the S. coelicolor ⌬wblA mutant produces a much higher level of actinorhodin than S. coelicolor M145 (29,30), whereas the adpA-deleted S. coelicolor strain does not produce actinorhodin at all in either liquid or plate cultures (15,16).…”

mentioning

confidence: 99%

“…To test this, transcription analysis was conducted with strains of S. coelicolor M145, the S. coelicolor ⌬wblA mutant (26,29), and the S. coelicolor ⌬adpA mutant (19). It was previously reported that the S. coelicolor ⌬wblA mutant produces a much higher level of actinorhodin than S. coelicolor M145 (29,30), whereas the adpA-deleted S. coelicolor strain does not produce actinorhodin at all in either liquid or plate cultures (15,16). Each strain was grown in 100 ml of modified R5 (R5 without KH 2 PO 4 , CaCl 2 · 2H 2 O, L-proline, and growth factor) liquid medium, and total RNAs were isolated (RNeasy miniprep kit; Qiagen) at 8 h (before any antibiotic production from all strains), 25 h (when there is a slight production of actinorhodin from just the ⌬wblA mutant), and 35 h (when there is the production of actinorhodin from S. coelicolor M145 or the ⌬wblA mutant and no production of actinorhodin from the ⌬adpA mutant).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Repression of Antibiotic Downregulator WblA by AdpA in Streptomyces coelicolor

Lee

Kim

et al. 2013

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

supporting

confidence: 59%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Repression of Antibiotic Downregulator WblA by AdpA in Streptomyces coelicolor

Lee

Kim

et al. 2013

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…These data indicated that the effect of overexpressing zwf2 was slightly small when the three genes were overexpressed (Table S1). This might be due to the imperfect nature of the in silico metabolic model that could not account for complex regulatory mechanisms (Kim et al 2011). Besides, another reason might be the synergistic effect caused by the adjacent dptI and dptJ genes in the genome of S. roseosporus.…”

Section: Discussionmentioning

confidence: 99%

In silico aided metabolic engineering of Streptomyces roseosporus for daptomycin yield improvement

Huang

Wen

Wang

et al. 2012

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

In silico metabolic network models are valuable tools for strain improvement with desired properties. In this work, based on the comparisons of each pathway flux under two different objective functions for the reconstructed metabolic network of Streptomyces roseosporus, three potential targets of zwf2 (code for glucose-6-phosphate hydrogenase), dptI (code for α-ketoglutarate methyltransferase), and dptJ (code for tryptophan oxygenase) were identified and selected for the genetic modifications. Overexpression of zwf2, dptI, and dptJ genes increased the daptomycin concentration up to 473.2, 452.5, and 489.1 mg/L, respectively. Furthermore, co-overexpression of three genes in series resulted in a 34.4% higher daptomycin concentration compared with the parental strain, which ascribed to the synergistic effect of the enzymes responsible for daptomycin biosynthesis. Finally, the engineered strain enhanced the yield of daptomycin up to 581.5 mg/L in the fed-batch culture, which was approximately 43.2% higher than that of the parental strain. These results demonstrated that the metabolic network based on in silico prediction would be accurate, reasonable, and practical for target gene identification and strain improvement.

show abstract

“…Sequential targeted gene disruption of these two regulatory genes as well as SCO5426, a flux-controlling gene, resulted in a tripleknockout S. coelicolor mutant displaying significantly increased production of the polyketide antibiotic actinorhodin (15,16,46). To determine whether or not deletion of these regulatory genes could affect CsA hydroxylation activity in S. coelicolor, we introduced pMMBL312 into S. coelicolor mutants, generating S. coelicolor M145 ⌬SCO5426, S. coelicolor M145 ⌬wblA, and S. coelicolor M145 ⌬wblA ⌬SCO1712 ⌬SCO5426 (designated ESK616, ESK617, and ESK618, respectively).…”

Section: Resultsmentioning

confidence: 99%

Identification of a Cyclosporine-Specific P450 Hydroxylase Gene through Targeted Cytochrome P450 Complement (CYPome) Disruption in Sebekia benihana

Lee

Kim

Yoon

et al. 2013

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

c It was previously proposed that regio-specific hydroxylation of an immunosuppressive cyclosporine (CsA) at the 4th N-methyl leucine is mediated by cytochrome P450 hydroxylase (CYP) in the rare actinomycete Sebekia benihana. This modification is thought to be the reason for the hair growth-promoting side effect without the immunosuppressive activity of CsA. Through S. benihana genome sequencing and in silico analysis, we identified the complete cytochrome P450 complement (CYPome) of S. benihana, including 21 CYPs and their electron transfer partners, consisting of 7 ferredoxins (FDs) and 4 ferredoxin reductases (FDRs). Using Escherichia coli conjugation-based S. benihana CYPome-targeted disruption, all of the identified CYP, FD, and FDR genes in S. benihana were individually inactivated. Among the 32 S. benihana exconjugant mutants tested, only a single S. benihana CYP mutant, ⌬CYP-sb21, failed to exhibit CsA hydroxylation activity. The hydroxylation was restored by CYP-sb21 gene complementation. Since all S. benihana FD and FDR disruption mutants maintained CsA hydroxylation activity, it can be concluded that CYP-sb21, a new member of the bacterial CYP107 family, is the only essential component of the in vivo regio-specific CsA hydroxylation process in S. benihana. Moreover, expression of an extra copy of the CYP-sb21 gene increased CsA hydroxylation in wild-type S. benihana and an NADPH-enriched Streptomyces coelicolor mutant, by 2-fold and 1.5-fold, respectively. These results show for the first time that regio-specific hydroxylation of CsA is carried out by a specific P450 hydroxylase present in S. benihana, and they set the stage for the biotechnological application of regio-specific CsA hydroxylation through heterologous CYP-sb21 expression.

show abstract

Transcriptome Analysis of an Antibiotic Downregulator Mutant and Synergistic Actinorhodin Stimulation via Disruption of a Precursor Flux Regulator in Streptomyces coelicolor

Cited by 27 publications

References 23 publications

Repression of Antibiotic Downregulator WblA by AdpA in Streptomyces coelicolor

Repression of Antibiotic Downregulator WblA by AdpA in Streptomyces coelicolor

In silico aided metabolic engineering of Streptomyces roseosporus for daptomycin yield improvement

Identification of a Cyclosporine-Specific P450 Hydroxylase Gene through Targeted Cytochrome P450 Complement (CYPome) Disruption in Sebekia benihana

Contact Info

Product

Resources

About