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Evaluation of novel inducible promoter/repressor systems for recombinant protein expression in Lactobacillus plantarum

Overview of attention for article published in Microbial Cell Factories, March 2016
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1 Google+ user

Citations

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Title
Evaluation of novel inducible promoter/repressor systems for recombinant protein expression in Lactobacillus plantarum
Published in
Microbial Cell Factories, March 2016
DOI 10.1186/s12934-016-0448-0
Pubmed ID
Authors

Silvia Heiss, Angelika Hörmann, Christopher Tauer, Margot Sonnleitner, Esther Egger, Reingard Grabherr, Stefan Heinl

Abstract

Engineering lactic acid bacteria (LAB) is of growing importance for food and feed industry as well as for in vivo vaccination or the production of recombinant proteins in food grade organisms. Often, expression of a transgene is only desired at a certain time point or period, e.g. to minimize the metabolic burden for the host cell or to control the expression time span. For this purpose, inducible expression systems are preferred, though cost and availability of the inducing agent must be feasible. We selected the plasmid free strain Lactobacillus plantarum 3NSH for testing and characterization of novel inducible promoters/repressor systems. Their feasibility in recombinant protein production was evaluated. Expression of the reporter protein mCherry was monitored with the BioLector(®) micro-fermentation system. Reporter gene mCherry expression was compared under the control of different promoter/repressor systems: PlacA (an endogenous promoter/repressor system derived from L. plantarum 3NSH), PxylA (a promoter/repressor system derived from Bacillus megaterium DSMZ 319) and PlacSynth (synthetic promoter and codon-optimized repressor gene based on the Escherichia coli lac operon). We observed that PlacA was inducible solely by lactose, but not by non-metabolizable allolactose analoga. PxylA was inducible by xylose, yet showed basal expression under non-induced conditions. Growth on galactose (as compared to exponential growth phase on glucose) reduced basal mCherry expression at non-induced conditions. PlacSynth was inducible with TMG (methyl β-D-thiogalactopyranoside) and IPTG (isopropyl β-D-1-thiogalactopyranoside), but also showed basal expression without inducer. The promoter PlacSynth was used for establishment of a dual plasmid expression system, based on T7 RNA polymerase driven expression in L. plantarum. Comparative Western blot supported BioLector(®) micro-fermentation measurements. Conclusively, overall expression levels were moderate (compared to a constitutive promoter). We evaluated different inducible promoters, as well as an orthologous expression system, for controlled gene expression in L. plantarum. Furthermore, here we provide proof of concept for a T7 RNA polymerase based expression system for L. plantarum. Thereby we expanded the molecular toolbox for an industrial relevant and generally regarded as safe (GRAS) strain.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 82 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Mexico 1 1%
China 1 1%
Austria 1 1%
Unknown 79 96%

Demographic breakdown

Readers by professional status Count As %
Researcher 17 21%
Student > Bachelor 16 20%
Student > Ph. D. Student 11 13%
Student > Master 8 10%
Other 6 7%
Other 16 20%
Unknown 8 10%
Readers by discipline Count As %
Agricultural and Biological Sciences 30 37%
Biochemistry, Genetics and Molecular Biology 28 34%
Immunology and Microbiology 4 5%
Engineering 3 4%
Pharmacology, Toxicology and Pharmaceutical Science 2 2%
Other 3 4%
Unknown 12 15%

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 19 November 2017.
All research outputs
#6,987,130
of 12,159,148 outputs
Outputs from Microbial Cell Factories
#447
of 884 outputs
Outputs of similar age
#134,922
of 284,523 outputs
Outputs of similar age from Microbial Cell Factories
#11
of 29 outputs
Altmetric has tracked 12,159,148 research outputs across all sources so far. This one is in the 40th percentile – i.e., 40% of other outputs scored the same or lower than it.
So far Altmetric has tracked 884 research outputs from this source. They receive a mean Attention Score of 3.7. This one is in the 46th percentile – i.e., 46% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 284,523 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 49th percentile – i.e., 49% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 29 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 55% of its contemporaries.