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General Information

Document type
  • Journal article
GE organism
  • soybean
GE trait
  • herbicide tolerance
Country
  • Korea, USA

Results

Safety for consumption
  • no effect

The 5-enolpyruvylshikimate-3-phosphate synthase of glyphosate-tolerant soybean expressed in Escherichia coli shows no severe allergenicity

Chang, HS; Kim, NH; Park, MJ; Lim, SK; Kim, SC; Kim, JY; Kim, JA; Oh, HY; Lee, CH; Huh, K; Jeong, TC; Nam, DH
Molecules and Cells. 2003 February . 15(1):20-26

Link to full text (journal may charge for access)

PMID: 12661756

Abstract

The recombinant gene was amplified from the chromosomal DNA of genetically-modified (GM) soybeans and identified as epsps encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) which renders glyphosate resistance. The epsps structural gene was introduced in the pET28(a) plasmid for its expression in Escherichia coli BL21(DE3). It was confirmed that the maximal productivity of the EPSPS protein was achieved when cultivating the recombinant strain in a LB broth for 2 h after supplementing 1 mM isopropylbeta-D-thiogalactopyranoside (IPTG) in a 2 h-culture broth. Since the expressed EPSPS protein was found as an insoluble form in the inclusion body, it was extracted by 6 M urea after sonication, and then purified through immobilized nickel-affinity column chromatography to isolate EPSPS having a molecular mass of 57 kDa. When incubated in simulated gastric fluid containing pepsin at pH 1.5, the purified EPSPS protein was completely digested within 1 min. In addition, the passive cutaneous anaphylaxis reaction of the purified EPSPS protein was not observed in the Sprague Dawley rat system that was administered either orally or subcutaneously. Furthermore, treatment of the EPSPS protein to the culture of the sensitized peritoneal mast cells, or unsensitized but antisera-labeled mast cells, showed neither a remarkable change in the histamine release nor a cytokine production, including interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). Thus, it can be concluded that the EPSPS protein in the GM soybean showed no significant allergenicity in the Sprague Dawley rats.

Keywords

Enolpyruvylshikimate-3-Phosphate Synthase; Allergenicity; Cytokine Production; Gene Expression; Genetically-Modified Soybean; Glyphosate-tolerant Soybean; Histamine Release; Passive Cutaneous Anaphylaxis; Pepsin Digestion

Funding

Funding source
  • Korea Food and Drug Administration
Funding country
  • Korea
Funding type
  • government

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Cite this study

MLA

Chang, HS, NH Kim, MJ Park, SK Lim, SC Kim, JY Kim, JA Kim, HY Oh, CH Lee, K Huh, TC Jeong, DH Nam. "The 5-enolpyruvylshikimate-3-phosphate synthase of glyphosate-tolerant soybean expressed in Escherichia coli shows no severe allergenicity ." Molecules and Cells 15.1 (2003): 20-26 . Web. 25 Sep. 2017.

APA

Chang, HS., Kim, NH., Park, MJ., Lim, SK., Kim, SC., Kim, JY., Kim, JA., Oh, HY., Lee, CH., Huh, K., Jeong, TC., & Nam, DH. (2003). The 5-enolpyruvylshikimate-3-phosphate synthase of glyphosate-tolerant soybean expressed in Escherichia coli shows no severe allergenicity . Molecules and Cells, 15(1), 20-26 .

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