Enhancement of doxorubicin production by expression of structural sugar biosynthesis and glycosyltransferase genes in Streptomyces peucetius

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

doi:10.1016/j.jbiosc.2009.03.002

Cited by 36 publications

(22 citation statements)

References 19 publications

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“…The combination of bldA and dnrO was cloned in pIBR25 vector using XbaI and HindIII site. The rDNAs were demethylated using E. coli ET 12567 host for protoplast transformation in S. peucetius ATCC 27952 (Malla et al 2009). …”

Section: Construction Of Recombinant Plasmidsmentioning

confidence: 99%

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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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Section: Construction Of Recombinant Plasmidsmentioning

confidence: 99%

“…peucetius and its variants were grown in NDYE medium (Malla et al 2009). Maximum mycelial density (OD600) was observed at 48 h (Fig.…”

Section: Growth Rate Analysis and Dnr Productionmentioning

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 polymerase chain reaction (PCR) product was cloned into pSET152 at the BamHI/XbaI sites to construct pDK1609 integration recombinant plasmid. For overexpression of dnrQS and drrAB genes, recombinant expression plasmids pDnrQS25 and pDrrAB25 were constructed by Malla et al [11,12], respectively. The PCR products were cloned into pGEM-T Easy and sequenced prior to cloning into the integration vector to verify that no mutation had occurred during PCR amplification.…”

Section: Construction Of Recombinant Plasmidsmentioning

confidence: 99%

“…The conditions for protoplast formation, regeneration, and DNA transformation were as we described previously [11]. To select S. peucetius transformants harboring pDK1609, 60 lg ml -1 apramycin was applied.…”

Section: Construction Of Recombinant Plasmidsmentioning

confidence: 99%

Proteomic approach to enhance doxorubicin production in panK-integrated Streptomyces peucetius ATCC 27952

Song

Malla

Yang

et al. 2011

J Ind Microbiol Biotechnol

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Biosynthesis of polyketide compounds depends upon the starter and extender units of coenzyme A derivatives of carboxylic acids present in the host organism. To increase the coenzyme A (CoA) pool, pantothenate kinase (panK) gene from Escherichia coli was integrated into S. peucetius ATCC 27952 (panK-integrated strain, BG200), which resulted in increase in aglycone polyketide ε-rhodomycinone (RHO), but decrease in the desired product, i.e., doxorubicin (DXR). To reduce RHO accumulation by synthesizing daunorubicin (DNR) from RHO more efficiently, glycosyltransferase (dnrQS) was overexpressed (pIBR25::dnrQS in panK-integrated strain, BG201). However, DnrQS overexpression still resulted in less production of DXR compared with the parental strain. To understand the results in detail by investigating the proteome changes in the panK-integrated strain, two-dimensional (2D) gel electrophoresis was performed. Among the several proteins that are up- or downregulated in BG200, efflux protein DrrA was our main target of interest, because it is directly related to DXR/DNR production in S. peucetius. DXR transporter DrrAB was additionally introduced in BG200 to enhance secretion of toxic DXR. Compared with S. peucetius ATCC 27952, BG204 (pIBR25::drrAB in panK-integrated strain), produced two times higher amount of DXR, which is 9.4-fold higher than that of panK-integrated strain BG200. The results show that the proteomic approach is quite useful in host development of Streptomyces and understanding cell physiology for antibiotic production.

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“…4,5 The research on daunorubicin and doxorubicin is now under renaissance, in particular with the studies carried by Professors Sohng (Sun Moon University, Korea) and Prasad (Madurai Kamaraj University, India) and their collaborators, published in the last few years. [9][10][11][12][13][14] This time, we have accumulated a huge knowledge about actinomycetes and the secondary metabolites produced by these microorganisms. A large number of biosynthetic gene clusters have been isolated and numerous novel compounds have been developed in the last 25 years in this field of research.…”

Section: Introductionmentioning

confidence: 99%

Hutchinson's legacy: keeping on polyketide biosynthesis

Olano

2010

J Antibiot

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Professor Charles Richard Hutchinson (Hutch) dedicated his research to the study of polyketide compounds, in particular, those produced by actinomycetes. Hutch principally centered his efforts to study the biosynthesis of bioactive compounds, antibiotic and antitumor drugs, and to develop new derivatives with improved therapeutic properties. After dedicating 40 years to the study of polyketides, Hutch leaves us, as legacy, the knowledge that he and his collaborators have accumulated and shared with the scientific community. The best tribute we can offer to him is keeping on the study of polyketides and other bioactive compounds, in an effort to generate more safer and useful drugs. In this review, the work on the polyketides, borrelidin, steffimycin and streptolydigin, performed at the laboratory of Professors Salas and Mé ndez at University of Oviedo (Spain) during the last 10 years, is summarized.

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Enhancement of doxorubicin production by expression of structural sugar biosynthesis and glycosyltransferase genes in Streptomyces peucetius

Cited by 36 publications

References 19 publications

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

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

Proteomic approach to enhance doxorubicin production in panK-integrated Streptomyces peucetius ATCC 27952

Hutchinson's legacy: keeping on polyketide biosynthesis

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