Skip to main content
SpringerLink
Log in

Targeting the middle region of CP4-EPSPS protein for its traceability in highly processed soy-related products

  • Original Article
  • Published:
Journal of Food Science and Technology Aims and scope Submit manuscript

Abstract

Transgenic components in genetically modified organisms consist not only of the transgenic genes, but also the transgenic protein. However, compared with transgenic DNA, less attention has been paid to the detection of expressed protein, especially those degraded from genetically modified soybean after food processing. In this study, the full length 5-enolpyruvyl-shikimate-3-phosphate synthase (CP4-EPSPS, 47.6 kD) protein was probed with the SC-16 (S19–R33) and the DC-16 (D219–K233) polyclonal antibodies in immunoblots. Both antibodies were able to detect the full length CP4-EPSPS and its residues in soy powder made from Roundup-Ready soybeans after heating and microwaving treatments which also reduced the molecular weight of the protein to 45.8 and 38.7 kD, respectively. Taken together the immunoblot results suggest that the middle region of the CP4-EPSPS protein possessed better stability than its N-terminal during thermal processing. This deduction was further validated by autoclave treatment, where a 37.4 kD residue of the protein was recognized by DC-16. A similar result was obtained in processed smoked sausage containing Roundup Ready soybean protein isolate (as an extender). The additional use of a further polyclonal antibody CK-17 (C372–K388), showed that compared with only the one signal for CP4-EPSPS detected by the SC-16 and CK-17 antibodies, the DC-16 middle region antibody detected four signals for CP4-EPSPS from five market sourced soy protein concentrates. Taken together, the study suggested that the middle region of CP4-EPSPS was more useful than the N- and C-terminal for tracing transgenic CP4-EPSPS protein and its remnants in highly processed soy-related products.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ahmed FE (2002) Detection of genetically modified organisms in foods. Trend Biotechnol 20:215–223

    Article  CAS  Google Scholar 

  • Ballari RV, Martin A (2013) Assessment of DNA degradation induced by thermal and UV radiation processing: implications for quantification of genetically modified organisms. Food Chem 141:2130–2136

    Article  CAS  Google Scholar 

  • Bogani P, Minunni M, Spiriti MM, Zavaglia M, Tombelli S, Buiatti M, Mascini M (2009) Transgenes monitoring in an industrial soybean processing chain by DNA-based conventional approaches and biosensors. Food Chem 113:658–664

    Article  CAS  Google Scholar 

  • Bøhn T, Cuhra M, Traavik T, Sanden M, Fagan J, Primicerio R (2014) Compositional differences in soybeans on the market: glyphosate accumulates in Roundup Ready GM soybeans. Food Chem 153:207–215

    Article  Google Scholar 

  • Branquinho MR, Ferreira RTB, Cardarelli-Leite P (2010) Survey of compliance with labeling legislation in food containing GMOs in Brazil. J Food Compos Anal 23:220–225

    Article  Google Scholar 

  • Brett GM, Chambers SJL, Huang L, Morgan MRA (1999) Design and development of immunoassays for detection of proteins. Food Control 10:401–406

    Article  Google Scholar 

  • Charleston B, Fray MD, Baigent S, Carr BV, Morrison WI (2001) Establishment of persistent infection with non-cytopathic bovine viral diarrhoea virus in cattle is associated with a failure to induce type I interferon. J Gen Virol 82:1893–1897

    Article  CAS  Google Scholar 

  • Chen H, Lin YG (2013) Promise and issues of genetically modified crops. Curr Opin Plant Biol 16:255–260

    Article  Google Scholar 

  • Chen Y, Wang Y, Ge Y, Xu B (2005) Degradation of endogenous and exogenous genes of Roundup-Ready soybean during food processing. J Agric Food Chem 53:10239–10243

    Article  CAS  Google Scholar 

  • de Luis R, Lavilla M, Sánchez L, Calvo M, Pérez MD (2009) Immunochemical detection of Cry1A(b) protein in model processed foods made with transgenic maize. Eur Food Res Technol 229:15–19

    Article  CAS  Google Scholar 

  • Gasser CS, Winter JA, Hironaka CM, Shah DM (1988) Structure, expression, and evolution of the 5-enolpyruvylshikimate-3-phosphate synthase genes of petunia and tomato. J Biol Chem 263:4280–4287

    CAS  Google Scholar 

  • Hammond BG, Jez JM (2011) Impact of food processing on the safety assessment for proteins introduced into biotechnology-derived soybean and corn crops. Food Chem Toxicol 49:711–721

    Article  CAS  Google Scholar 

  • Holst-Jensen A, Bertheau Y, de Loose M, Grohmann L, Hamels S, Hougs L, Morisset D, Pecoraro S, Pla M, Van den Bulcke M, Wulff D (2012) Detecting un-authorized genetically modified organisms (GMOs) and derived materials. Biotechnol Adv 30:1318–1335

    Article  CAS  Google Scholar 

  • Huang M, Shen W, Wu H, Xiao X, Xu S, Zhou X, He J, Zhou G (2012) Polyclonal antibody performing specific antigen-antibody reaction with CP4-EPSPS protein and application thereof. Patent, CN ZL 201010225353.3

  • Kinsella JE (1979) Functional properties of soy protein. J Am Oil Chem Soc 56:242–258

    Article  CAS  Google Scholar 

  • Nida DL, Kolacz KH, Buehler RE, Randy Deaton W, Schler WR, Armstrong TA, Taylor ML, Ebert CC, Rogan GJ, Padgette SR, Fuchs RL (1996) Glyphosate-tolerant cotton: genetic characterization and protein expression. J Agric Food Chem 44:1960–1966

    Article  CAS  Google Scholar 

  • Onori R, Lopardo R, de Giacomo M, de Santis B, Prantera E, Palmaccio E, Brera C (2013) Traceability of genetically modified Roundup Ready soybean: a case study on sampling and analytical uncertainty along processing chain. Food Control 34:494–501

    Article  CAS  Google Scholar 

  • Notification No. 2003-31 (2000) Ministry of Agriculture and Forestry of Korea, Seoul, Korea, April 22, 2000

  • Notification No. 1775 (2000) Food and Marketing Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan, Tokyo, Japan, June 10, 2000

  • Rogan GJ, Dubin YA, Lee TC, Magin KM, Astwood JD, Bhakta NS, Leach JN, Sanders PR, Fuchs RL (1999) Immunodiagnostic methods for detection of 5-enolpyruvylshikimate-3-phosphate synthase in Roundup Ready® soybeans. Food Control 10:407–414

    Article  Google Scholar 

  • Sadeghi AA, Nikkhah A, Shawrang P, Shahrebabak MM (2006) Protein degradation kinetics of untreated and treated soybean meal using SDS-PAGE. Anim Feed Sci Technol 12:122–131

    Google Scholar 

  • Smales MC, Pepper DS, James DC (2002) Protein modification during anti-viral heat-treatment bioprocessing of factor VIII concentrates, factor IX concentrates and model proteins in the presence of sucrose. Biotechnol Bioeng 77:37–48

    Article  CAS  Google Scholar 

  • Tian F, Guan Q, Wang X, Teng D, Wang J (2014) Influence of different processing treatments on the detectability of nucleic acid and protein targets in transgenic soybean meal. Appl Biochem Biotechnol 172:3686–3700

    Article  CAS  Google Scholar 

  • Tsumura K, Saito T, Tsuge K, Ashida H, Kugimiya W, Inouye K (2005) Functional properties of soy protein hydrolysates obtained by selective proteolysis. LWT-Food Sci Technol 38:255–261

    Article  CAS  Google Scholar 

  • Ueda K, Fukushima H, Masliah E, Xia Y, Iwai A, Yoshimoto M, Otero DAC, Kondo J, Ihara Y, Saitoh T (1993) Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. Proc Natl Acad Sci USA 90:11282–11286

    Article  CAS  Google Scholar 

  • Uzogara SG (2000) The impact of genetic modification of human foods in the 21st century: a review. Biotechnol Adv 18:179–206

    Article  CAS  Google Scholar 

  • van Duijn G, van Biert R, Bleeker-Marcelis H, Peppelman H, Hessing M (1999) Detection methods for genetically modified crops. Food Control 10:375–378

    Article  Google Scholar 

  • van Remmen HHJ, Ponne CT, Nijhuis HH, Bartels PV, Kerkhof PJAM (1996) Microwave heating distributions in slabs, spheres and cylinders with relation to food processing. J Food Sci 61:1105–1114

    Article  Google Scholar 

  • Vijayakumar KR, Martin A, Gowda LR, Prakash V (2009) Detection of genetically modified soya and maize: impact of heat processing. Food Chem 117:514–521

    Article  CAS  Google Scholar 

  • Wu HH, Zhang Y, Zhu CQ, Xiao X, Zhou XH, Xu S, Shen WB, Huang M (2012) Presence of CP4-EPSPS component in Roundup Ready soybean-derived food products. Int J Mol Sci 13:1919–1932

    Article  CAS  Google Scholar 

  • Xiao X, Wu HH, Zhou XH, Xu S, He J, Shen WB, Zhou GH, Huang M (2012) The combination of quantitative PCR and western blot detecting CP4-EPSPS component in Roundup Ready Soy plant tissues and commercial Soy-related foodstuffs. J Food Sci 77:C603–C608

    Article  CAS  Google Scholar 

  • Yu P (2005) Protein secondary structures (α-helix and β-sheet) at a cellular level and protein fractions in relation to rumen degradation behaviours of protein: a new approach. Br J Nutr 94:655–665

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. Sheng Xu, and Mrs. Xiao Xiao from Nanjing Agricultural University for the technical support. We thank Dr. Evan Evans and Mr. Joseph Hartley from University of Tasmania, Australia for the critical reading and comments on this manuscript. We thank the anonymous reviewer for the insightful suggestions. This work was supported by the National Transgenic Major Program (No. 2009ZX08012-014B), and the National Natural Science Foundation of China (J1210056 and J1310015).

Author information

Author notes

  1. Honghong Wu

    Present address: Department of Botany and Plant Sciences, University of California at Riverside, Riverside, CA, 92521, USA

  2. Xinghu Zhou

    Present address: Nanjing Agricultural University Meat & Food Co. Ltd., Nanjing, 210095, China

  3. Honghong Wu, Xiaofu Wang, Xinghu Zhou these authors contributed equally to this work.

Authors and Affiliations

  1. College of Life Sciences, Laboratory Center of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China

    Honghong Wu, Yihua Zhang, Jian He & Wenbiao Shen

  2. Institute of Agriculture Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China

    Xiaofu Wang

  3. College of Food Sciences, National Center of Meat Quality and Safety Control, Nanjing Agricultural University, Nanjing, 210095, China

    Xinghu Zhou & Ming Huang

Authors
  1. Honghong Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaofu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xinghu Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yihua Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ming Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jian He
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wenbiao Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wenbiao Shen.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 1463 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wu, H., Wang, X., Zhou, X. et al. Targeting the middle region of CP4-EPSPS protein for its traceability in highly processed soy-related products. J Food Sci Technol 54, 3142–3151 (2017). https://doi.org/10.1007/s13197-017-2750-1

Download citation

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13197-017-2750-1

Keywords

Search

Navigation