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Biomolecules2018; 8(4); 146; doi: 10.3390/biom8040146

Induction of Recombinant Lectin Expression by an Artificially Constructed Tandem Repeat Structure: A Case Study Using Bryopsis plumosa Mannose-Binding Lectin.

Abstract: Lectin is an important protein in medical and pharmacological applications. Impurities in lectin derived from natural sources and the generation of inactive proteins by recombinant technology are major obstacles for the use of lectins. Expressing recombinant lectin with a tandem repeat structure can potentially overcome these problems, but few studies have systematically examined this possibility. This was investigated in the present study using three distinct forms of recombinant mannose-binding lectin from (BPL2)-i.e., the monomer (rD1BPL2), as well as the dimer (rD2BPL2), and tetramer (rD4BPL2) arranged as tandem repeats. The concentration of the inducer molecule isopropyl β-D-1-thiogalactopyranoside and the induction time had no effect on the efficiency of the expression of each construct. Of the tested constructs, only rD4BPL2 showed hemagglutination activity towards horse erythrocytes; the activity of towards the former was 64 times higher than that of native BPL2. Recombinant and native BPL2 showed differences in carbohydrate specificity; the activity of rD4BPL2 was inhibited by the glycoprotein fetuin, whereas that of native BPL2 was also inhibited by d-mannose. Our results indicate that expression as tandem repeat sequences can increase the efficiency of lectin production on a large scale using a bacterial expression system.
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Publication Date: 2018-11-14 PubMed ID: 30441842PubMed Central: PMC6316659DOI: 10.3390/biom8040146Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research explores how producing a recombinant lectin protein, using an artificially created tandem repeat structure, can improve the efficiency of lectin production. The study shows that this method significantly enhanced the activity of the resulting lectin compared to a naturally derived source.

Objective of the Research

  • The main objective of this research was to investigate whether expressing recombinant lectin with an artificially made tandem repeat structure could overcome the main limitations encountered in the use of lectins derived from natural sources or generated through recombinant technology.
  • A particular interest was in understanding the impact of this artificial tandem repeat structure on the production efficiency and biological activity of the recombinant lectin.

Methods and Analysis

  • The researchers used three distinct forms of recombinant mannose-binding lectin (BPL2): a monomer (rD1BPL2), a dimer (rD2BPL2), and a tetramer (rD4BPL2). These different forms were arranged as tandem repeats.
  • The study examined the effect of the concentration of the inducer molecule isopropyl β-D-1-thiogalactopyranoside and the induction time on the efficiency of expression of each construct.
  • Additionally, they compared the hemagglutination activity of the recombinant constructs towards horse erythrocytes and the carbohydrate specificity differences between recombinant and native BPL2.

Findings

  • The study found that the concentration of the inducer molecule and the induction time did not affect the efficiency of the expression for the different constructs.
  • Out of the tested constructs, only rD4BPL2 showed hemagglutination activity towards horse erythrocytes. This activity was reported to be significantly higher than that of native BPL2, indicating that the recombinant method was more effective.
  • Notably, recombinant and native BPL2 showed differences in carbohydrate specificity. The activity of rD4BPL2 was inhibited by the glycoprotein fetuin, while the activity of native BPL2 was also inhibited by d-mannose.

Conclusions

  • The results from this study suggest that using tandem repeat sequences can increase the efficiency of production of lectin on a large scale. This method could improve the effectiveness and potential applications of lectin in medical and pharmacological fields.

Cite This Article

APA
Hwang HJ, Han JW, Jeon H, Han JW. (2018). Induction of Recombinant Lectin Expression by an Artificially Constructed Tandem Repeat Structure: A Case Study Using Bryopsis plumosa Mannose-Binding Lectin. Biomolecules, 8(4), 146. https://doi.org/10.3390/biom8040146

Publication

Biomolecules
ISSN: 2218-273X
NlmUniqueID: 101596414
Country: Switzerland
Language: English
Volume: 8
Issue: 4
PII: 146

Researcher Affiliations

Hwang, Hyun-Ju
  • Department of Genetic Resources, National Marine Biodiversity Institute of Korea, Seocheon 33662, Korea. hjhwang@mabik.re.kr.
Han, Jin-Woo
  • Department of Genetic Resources, National Marine Biodiversity Institute of Korea, Seocheon 33662, Korea. hiclow@mabik.re.kr.
Jeon, Hancheol
  • Department of Genetic Resources, National Marine Biodiversity Institute of Korea, Seocheon 33662, Korea. hjeon@mabik.re.kr.
Han, Jong Won
  • Department of Genetic Resources, National Marine Biodiversity Institute of Korea, Seocheon 33662, Korea. jwhan@mabik.re.kr.

MeSH Terms

  • Amino Acid Sequence
  • Animals
  • Carbohydrates / chemistry
  • Chlorophyta / chemistry
  • Hemagglutination Tests
  • Horses
  • Mannose-Binding Lectin / chemistry
  • Mannose-Binding Lectin / isolation & purification
  • Mannose-Binding Lectin / metabolism
  • Plant Lectins / chemistry
  • Plant Lectins / isolation & purification
  • Plant Lectins / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sheep
  • Solubility
  • Tandem Repeat Sequences

Conflict of Interest Statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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Citations

This article has been cited 3 times.
  1. Barre A, Van Damme EJM, Simplicien M, Le Poder S, Klonjkowski B, Benoist H, Peyrade D, Rougé P. Man-Specific Lectins from Plants, Fungi, Algae and Cyanobacteria, as Potential Blockers for SARS-CoV, MERS-CoV and SARS-CoV-2 (COVID-19) Coronaviruses: Biomedical Perspectives. Cells 2021 Jun 28;10(7).
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  2. Barre A, Damme EJMV, Simplicien M, Benoist H, Rougé P. Man-Specific, GalNAc/T/Tn-Specific and Neu5Ac-Specific Seaweed Lectins as Glycan Probes for the SARS-CoV-2 (COVID-19) Coronavirus. Mar Drugs 2020 Oct 29;18(11).
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  3. Hwang HJ, Han JW, Jeon H, Cho K, Kim JH, Lee DS, Han JW. Characterization of a Novel Mannose-Binding Lectin with Antiviral Activities from Red Alga, Grateloupia chiangii. Biomolecules 2020 Feb 19;10(2).
    doi: 10.3390/biom10020333pubmed: 32092955google scholar: lookup
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