Bioprocess intensification of antibiotic production by Streptomyces coelicolor A3(2) in micro-porous culture

Ndlovu, T.M.; Ward, Alan C.; Glassey, J.; Eskildsen, J.; Akay, G.

doi:10.1016/j.msec.2015.01.052

Cited by 15 publications

(25 citation statements)

References 42 publications

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“…Streptomyces lividans , a useful Gram-positive secretion model, can secrete several heterologous polypeptides of bacterial and eukaryotic origin (Pozidis et al, 2001; Hong et al, 2003; Lara et al, 2004; Ogino et al, 2004; Sianidis et al, 2006; Hamed et al, 2017; Kashiwagi et al, 2017). The absence of lipopolysaccharides, the availability of advanced genetic tools (Kieser et al, 2000; Kashiwagi et al, 2017), low protease activity, established industrial bioprocessing as a major producer of antibiotics (Ndlovu et al, 2015), synthetic biology tools (Phelan et al, 2017) and the avoidance of inclusion body formation, renders S. lividans secretion an attractive biotechnology platform. In many instances, it can provide alternative solutions when established workhorses, like Escherichia coli , fail (Anné et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Monitoring Protein Secretion in Streptomyces Using Fluorescent Proteins

Hamed

Vrancken

Bilyk³

et al. 2018

Front. Microbiol.

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Fluorescent proteins are a major cell biology tool to analyze protein sub-cellular topology. Here we have applied this technology to study protein secretion in the Gram-positive bacterium Streptomyces lividans TK24, a widely used host for heterologous protein secretion biotechnology. Green and monomeric red fluorescent proteins were fused behind Sec (SPSec) or Tat (SPTat) signal peptides to direct them through the respective export pathway. Significant secretion of fluorescent eGFP and mRFP was observed exclusively through the Tat and Sec pathways, respectively. Plasmid over-expression was compared to a chromosomally integrated spSec-mRFP gene to allow monitoring secretion under high and low level synthesis in various media. Fluorimetric detection of SPSec-mRFP recorded folded states, while immuno-staining detected even non-folded topological intermediates. Secretion of SPSec-mRFP is unexpectedly complex, is regulated independently of cell growth phase and is influenced by the growth regime. At low level synthesis, highly efficient secretion occurs until it is turned off and secretory preforms accumulate. At high level synthesis, the secretory pathway overflows and proteins are driven to folding and subsequent degradation. High-level synthesis of heterologous secretory proteins, whether secretion competent or not, has a drastic effect on the endogenous secretome, depending on their secretion efficiency. These findings lay the foundations of dissecting how protein targeting and secretion are regulated by the interplay between the metabolome, secretion factors and stress responses in the S. lividans model.

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Section: Introductionmentioning

confidence: 99%

Monitoring Protein Secretion in Streptomyces Using Fluorescent Proteins

Hamed

Vrancken

Bilyk³

et al. 2018

Front. Microbiol.

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“…Recently, porous structures of silicates, synthetic organic polymers, and natural (alginates) have been used as matrices for bacterial cell immobilization to improve the bioprocess productivity. In fact, besides reducing cell–cell aggregation, porous scaffolds increase the surface potentially available for cell adhesion, thus facilitating exchanges with the cultivation medium . The degree of microbial cell immobilization in porous structures depends on different factors, including the structure and the size of adsorbent pores.…”

Section: Introductionmentioning

confidence: 99%

Effect of PCL/PEG‐Based Membranes on Actinorhodin Production in Streptomyces coelicolor Cultivations

Scaffaro

Lopresti

Sutera

et al. 2016

Macromolecular Bioscience

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The actinomycetes, Gram-positive filamentous bacteria, are the most prolific source of natural occurring antibiotics. At an industrial level, antibiotics from actinomycete strains are produced by means of submerged fermentations, where one of the major factors negatively affecting bioproductivity is the pellet-shaped biomass growth. The immobilization of microorganisms on properly chosen supports prevents cell-cell aggregation resulting in improving the biosynthetic capability. Thus, novel porous biopolymer-based devices are developed by combining melt mixing and particulate leaching. In particular, polycaprolactone (PCL), polyethylene glycol (PEG), and sodium chloride (NaCl) with different grain sizes are used to prepare PCL/PEG/NaCl blends in the melt. These blends are then leached to obtain PCL-based porous membranes that are used as solid support for the growth of Streptomyces coelicolor, a model streptomycete used to produce various antibiotics including the blue colored actinorhodin (ACT). Thereafter, the effect of the devices' characteristics on the bacterial growth and on the production ACT is evaluated. The results showed that ACT production is strongly dependent on the pore size distribution of the device. Moreover, membranes with pores ranging from 90 to 110 μm are able to offer a potential improvement in volumetric productivity of ACT if compared to conventional submerged liquid culture.

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“…Ribosome recycling factor (RRF) is the protein that takes charge of dissociating the relevant RNA from ribosome when the transcription is completed [25,26]. For instance, over-expressing the gene frr in Streptomyces diastatochromo-genes 1628 has managed to increase the production of antibiotic [27,28]. RimP-SC is an assembly protein in ribosome, and knocking off this RimP-SC gene could increase the biosynthesis of antibiotic Act and CDA in Streptomyces coelicolor [29].…”

Section: Regulation Of Expression Of Ribosome-related Proteinsmentioning

confidence: 99%