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Soluble expression, purification, and characterization of active recombinant human tissue plasminogen activator by auto-induction in E. coli

Overview of attention for article published in BMC Biotechnology, March 2015
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About this Attention Score

  • In the top 25% of all research outputs scored by Altmetric
  • Among the highest-scoring outputs from this source (#46 of 382)
  • High Attention Score compared to outputs of the same age (84th percentile)
  • Average Attention Score compared to outputs of the same age and source

Mentioned by

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1 news outlet
twitter
1 tweeter

Citations

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14 Dimensions

Readers on

mendeley
43 Mendeley
Title
Soluble expression, purification, and characterization of active recombinant human tissue plasminogen activator by auto-induction in E. coli
Published in
BMC Biotechnology, March 2015
DOI 10.1186/s12896-015-0127-y
Pubmed ID
Authors

Xiaobin Long, Yeran Gou, Miao Luo, Shaocheng Zhang, Hongpeng Zhang, Lei Bai, Shuang Wu, Quan He, Ke Chen, Ailong Huang, Jianzhong Zhou, Deqiang Wang

Abstract

Human tissue plasminogen activator (tPA) belongs to the serine protease family. It converts plasminogen into plasmin and is used clinically to treat thrombosis. Human tPA is composed of 527 amino acids residues and contains 17 disulfide bonds. Escherichia coli has been used only rarely for the efficient production of recombinant tPA. However, the functional expression of full-length tPA that contains multiple disulfide bonds on an industrial scale remains challenging. Here, we describe the soluble expression and characterization of full-length tPA by auto-induction in E. coli. We achieved optimal levels of gene expression, minimized negative effects related to the production of heterologous proteins, and optimized cytoplasmic yields. Three different E. coli strains, BL21 (DE3), Rosetta, and Origami 2, could express tPA using an auto-induction mechanism. In addition, similar yields of recombinant protein were produced at temperatures of 33, 35, and 37°C. The E. coli strain origami 2 could increase disulfide bond formation in cytoplasmic tPA and produce purified soluble recombinant protein (~0.9 mg/l medium). The full-length tPA was monomeric in solution, and fibrin plate assays confirmed that the recombinant tPA displayed serine protease activity. This is the first report that describes the heterologous expression of correctly folded active full-length tPA. This could provide valuable information for using prokaryotic auto-induction expression systems to produce tPA at industrial and pharmaceutical levels without in vitro refolding during the production step.

Twitter Demographics

The data shown below were collected from the profile of 1 tweeter who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 43 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
China 1 2%
Canada 1 2%
Unknown 41 95%

Demographic breakdown

Readers by professional status Count As %
Student > Bachelor 12 28%
Student > Ph. D. Student 8 19%
Student > Master 6 14%
Researcher 4 9%
Unspecified 4 9%
Other 9 21%
Readers by discipline Count As %
Agricultural and Biological Sciences 16 37%
Biochemistry, Genetics and Molecular Biology 13 30%
Unspecified 4 9%
Medicine and Dentistry 2 5%
Engineering 2 5%
Other 6 14%

Attention Score in Context

This research output has an Altmetric Attention Score of 10. 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 01 March 2015.
All research outputs
#449,147
of 4,818,502 outputs
Outputs from BMC Biotechnology
#46
of 382 outputs
Outputs of similar age
#22,008
of 143,647 outputs
Outputs of similar age from BMC Biotechnology
#9
of 22 outputs
Altmetric has tracked 4,818,502 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 90th percentile: it's in the top 10% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 382 research outputs from this source. They receive a mean Attention Score of 4.2. This one has done well, scoring higher than 83% of its peers.
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 143,647 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 84% of its contemporaries.
We're also able to compare this research output to 22 others from the same source and published within six weeks on either side of this one. This one is in the 40th percentile – i.e., 40% of its contemporaries scored the same or lower than it.