Tools for metabolic engineering in<i>Streptomyces</i>

Bekker, Valerie; Dodd, Amanda; Brady, Dean; Rumbold, Karl

doi:10.4161/bioe.29935

Cited by 14 publications

(7 citation statements)

References 53 publications

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“…Advances in systems biological tools, such as next generation sequencing and "-omics" technologies, have led to more efficient ways to engineer microorganisms for secondary metabolite titer improvement (Bro and Nielsen, 2004;Bekker et al, 2014). Furthermore, the integration of genetic/metabolic engineering with systems biological tools provides invaluable contributions to increase secondary metabolite production (Hwang et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Genetic engineering of an industrial strain of Streptomyces clavuligerusfor further enhancement of clavulanic acid production

Kurt‐Kızıldoğan¹,

Jaccard²,

Mutlu³

et al. 2017

Turk J Biol

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An industrial clavulanic acid (CA) overproducer Streptomyces clavuligerus strain, namely DEPA, was engineered to further enhance its CA production. Single or multiple copies of ccaR, claR (pathway-specific activators), and cas2 (CA synthase) genes under the control of different promoters were introduced into this strain. CA titers of the resulting recombinants were analyzed by HPLC in a dynamic fashion and compared to the vector-only controls and a wild-type strain of S. clavuligerus while their growth was monitored throughout fermentation. The addition of an extra copy of ccaR, under control of its own promoter or constitutive ermE* promoter (P ermE* ), led to 7.6-and 2.3-fold increased volumetric levels of CA in respective recombinants, namely the AK9 and ID3 strains. Its highly stable multicopy expression by the glpF promoter (P glpF ) provided up to 25.9-fold enhanced volumetric CA titers in the respective recombinant, IDG3. claR expression controlled with its own promoter or ermE* and glpF-mediated amplification in an industrial strain brought about only about 1.2-fold increase in the volumetric CA titers. An extra copy of cas2 integration with P ermE* into the S. clavuligerus DEPA genome led to 3.8-fold higher volumetric CA production by GV61. Conclusively, multicopy expression of ccaR under P glpF can result in significantly improved industrial high-titer CA producers.

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Section: Discussionmentioning

confidence: 99%

Genetic engineering of an industrial strain of Streptomyces clavuligerusfor further enhancement of clavulanic acid production

Kurt‐Kızıldoğan¹,

Jaccard²,

Mutlu³

et al. 2017

Turk J Biol

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“…Electroporation involves the introduction of pores within bacterial membranes via an electric current to allow mass, unrestricted transfer of genetic material into species(52, 53). The efficiency of transformation, however, is species-dependent(52). Unlike electroporation, conjugation is not limited by the size of vectors that can be transformed, and has been used successfully to transfer entire genomes in E. coli (54).…”

Section: Discussionmentioning

confidence: 99%

“…Currently, the two most commonly used methods of bacterial gene transfer are conjugation and electroporation, both of which come with several advantages and disadvantages. Electroporation involves the introduction of pores within bacterial membranes via an electric current to allow mass, unrestricted transfer of genetic material into species(52, 53). The efficiency of transformation, however, is species-dependent(52).…”

Section: Discussionmentioning

confidence: 99%

Integrating Vectors for Genetic Studies in the Rare ActinomyceteAmycolatopsis marina

Gao

Murugesan

Hoßbach

et al. 2018

Preprint

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Few natural product pathways from rare Actinomycetes have been studied due to the difficulty in applying molecular approaches in these genetically intractable organisms. In this study, we sought to identify integrating vectors, derived using phage int/attP loci, that would efficiently integrate site-specifically in the rare Actinomycete, Amycolatopsis marina DSM45569. Analysis of the genome of A. marina DSM45569 indicated the presence of attB-like sequences for TG1 and R4 integrases. The TG1 and R4 attBs were active in in vitro recombination assays with their cognate purified integrases and attP loci. Integrating vectors containing either the TG1 or R4 int/attP loci yielded exconjugants in conjugation assays from E. coli to A. marina DSM45569. Site-specific recombination of the plasmids into the host TG1 or R4 attB sites was confirmed by sequencing. The presence of homologous TG1 and R4 attB sites in other species of this genus indicates that vectors based on TG1 and R4 integrases could be widely applicable.ImportanceRare Actinomycetes have the same potential of natural product discovery as Streptomyces, but the potential has not been fully explored due to the lack of efficient molecular biology tools. In this study, we identified two serine integrases, TG1 and R4, which could be used in the rare Actinomycetes species, Amycolatopsis marina, as tools for genome integration. The high level of conservation between the attB sites for TG1 and R4 in a number of Amycolatopsis species suggested that plasmids with the integration systems from these phages should be widely useful in this genus.

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“…Some nontraditional recent approaches, by which metabolic flux is amplified, and identified the genes through genome or transcriptome mining, concurrent expression outlining of numerous genes in plants, escapist flux through silencing and overexpression. Bekker et al [83] have summarized and discussed the published literature on tools for metabolic engineering of Streptomyces over the last decade. These strategies involve precursor engineering, structural and regulatory gene engineering, and the up or downregulation of genes, as well as genome shuffling and the use of genome scale metabolic models [3].…”

Section: Metabolic Bioengineering Toolsmentioning

confidence: 99%

Bioengineering tools for the production of pharmaceuticals: current perspective and future outlook

2019

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The bioengineering tools have significant advantages through less time-consuming and utilized as a promising stage for the production of pharmaceutical bioproducts under the single platform. This review highlighted the advantages and current improvement in the plant, animal and microbial bioengineering tools and outlines feasible approaches by biological and process's bioengineering levels for advancing the economic feasibility of pharmaceutical's production. The critical analysis results revealed that system biology and synthetic biology along with advanced bioengineering tools like transcriptome, proteome, metabolome and nano bioengineering tools have shown a promising impact on the development of pharmaceutical's bioproducts. Tools to overcome and resolve the accompanying encounters of pharmaceutical's production that include nano bioengineering tools are also discussed. As a summary and prospect, it also gives new insight into the challenges and possible breakthrough of the development of pharmaceutical's bioproducts through bioengineering tools.

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Tools for metabolic engineering inStreptomyces

Cited by 14 publications

References 53 publications

Genetic engineering of an industrial strain of Streptomyces clavuligerusfor further enhancement of clavulanic acid production

Genetic engineering of an industrial strain of Streptomyces clavuligerusfor further enhancement of clavulanic acid production

Integrating Vectors for Genetic Studies in the Rare ActinomyceteAmycolatopsis marina

Bioengineering tools for the production of pharmaceuticals: current perspective and future outlook

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