Improvement in doxorubicin productivity by overexpression of regulatory genes in Streptomyces peucetius

Malla, Sailesh; Niraula, Narayan Prasad; Liou, Kwangkyoung; Sohng, Jae Kyung

doi:10.1016/j.resmic.2009.12.003

Cited by 30 publications

(21 citation statements)

References 21 publications

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“…Hence, it is reasonable to suggest that otrC could be a valuable target for genetic manipulation, such as introducing extra copies of the ABC transporter [12,13], overexpressing the pathway genes with the use of transcription regulators [20-22] and the replacement of heterologous promoters [23,24], in order to yield the overproduction of OTC.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus

Yan

Wang

et al. 2012

BMC Biotechnol

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Background: The otrC gene of Streptomyces rimosus was previously annotated as an oxytetracycline (OTC) resistance protein. However, the amino acid sequence analysis of OtrC shows that it is a putative ATP-binding cassette (ABC) transporter with multidrug resistance function. To our knowledge, none of the ABC transporters in S. rimosus have yet been characterized. In this study, we aimed to characterize the multidrug exporter function of OtrC and evaluate its relevancy to OTC production. Results: In order to investigate OtrC's function, otrC is cloned and expressed in E. coli The exporter function of OtrC was identified by ATPase activity determination and ethidium bromide efflux assays. Also, the susceptibilities of OtrC-overexpressing cells to several structurally unrelated drugs were compared with those of OtrC-non-expressing cells by minimal inhibitory concentration (MIC) assays, indicating that OtrC functions as a drug exporter with a broad range of drug specificities. The OTC production was enhanced by 1.6-fold in M4018 (P = 0.000877) and 1.4-fold in SR16 (P = 0.00973) duplication mutants, while it decreased to 80% in disruption mutants (P = 0.0182 and 0.0124 in M4018 and SR16, respectively).

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Section: Discussionmentioning

confidence: 99%

Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus

Yan

Wang

et al. 2012

BMC Biotechnol

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“…One of the most widely employed promoters in streptomycetes is the erythromycin resistance gene ermEp from Saccharopolyspora erythraea or its upregulated variant ermEp ∗ [ 152 ]. For example, simultaneous overexpression of the dnrN , dnrI , and afsR regulatory genes under the control of ermEp ∗ in S. peucetius led to a 4.3-fold increase in doxorubicin production [ 153 ]. Another prominent example of the use of this promoter is the improvement in tylosin production in S. fradiae .…”

Section: Strain Improvement In the Postgenomic Eramentioning

confidence: 99%

Antibacterial Discovery and Development: From Gene to Product and Back

Fedorenko

Genilloud

Horbal

et al. 2015

BioMed Research International

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Concern over the reports of antibiotic-resistant bacterial infections in hospitals and in the community has been publicized in the media, accompanied by comments on the risk that we may soon run out of antibiotics as a way to control infectious disease. Infections caused by Enterococcus faecium, Staphylococcus aureus, Klebsiella species, Clostridium difficile, Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli, and other Enterobacteriaceae species represent a major public health burden. Despite the pharmaceutical sector's lack of interest in the topic in the last decade, microbial natural products continue to represent one of the most interesting sources for discovering and developing novel antibacterials. Research in microbial natural product screening and development is currently benefiting from progress that has been made in other related fields (microbial ecology, analytical chemistry, genomics, molecular biology, and synthetic biology). In this paper, we review how novel and classical approaches can be integrated in the current processes for microbial product screening, fermentation, and strain improvement.

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“…In addition, mutation in dnrO has also been shown to affect the expression of dnrN and dnrI (Otten et al 2000). An attempt has been made to overexpress dnrI and dnrN for production of DNR (Otten et al 1995a, b), however overexpression of dnrO has not been attempted for DNR production (Malla et al 2010). Interestingly, a closer look at dnrO, the key activator of DNR biosynthetic pathway of S. peucetius reveals the presence of a single TTA codon.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of a pathway specific negative regulator enhances production of daunorubicin in bldA deficient Streptomyces peucetius ATCC 27952

Pokhrel

Chaudhary

Nguyen

et al. 2016

Microbiological Research

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The dnrO gene is the first regulator to be activated in the daunorubicin (DNR) biosynthesis pathway of Streptomyces peucetius ATCC 27952. DnrO is known for its self-repression capability while it activates rest of the DNR biosynthesis pathway through cascades of regulatory events. S. peucetius was found to contain no functional copy of bldA-tRNA while a detailed examination of dnrO codons reveals the presence of TTA codon, which is rarely encoded by bldA-tRNA. Therefore, for evaluating the role of dnrO in DNR production, multiple engineered strains of S. peucetius were generated by heterologously expressing bldA, dnrO and combination of bldA and dnrO. Using these strains, the effects of heterologously expressed bldA and overexpressed dnrO were evaluated on pathway specific regulators, mycelial densities and production of DNR. The results showed that the transcription level of dnrO and master regulator dnrI, was found to be elevated in bldA containing strain in comparison to dnrO overexpressed strain. The bldA containing strain produces 45.7% higher DNR than bldA deficient wild type strain from culture broth with OD600 of 1.45 at 72h. Heterologous expression of bldA-tRNA is accounted for increased transcription levels of the DNR pathway specific regulators and enhanced DNR production.

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Improvement in doxorubicin productivity by overexpression of regulatory genes in Streptomyces peucetius

Cited by 30 publications

References 21 publications

Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus

Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus

Antibacterial Discovery and Development: From Gene to Product and Back

Overexpression of a pathway specific negative regulator enhances production of daunorubicin in bldA deficient Streptomyces peucetius ATCC 27952

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