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Safety Assessment of the Neomycin Phosphotransferase II (NPTII) Protein

Bio/Technology volume 11, pages 1543–1547 (1993)Cite this article

Abstract

Two approaches were used to assess the safety of the NPTII protein for human consumption using purified E. coli produced NPTII protein that was shown to be chemically and functionally equivalent to the NPTII protein produced in genetically engineered cotton seed, potato tubers and tomato fruit. The NPTII protein was shown, as expected, to degrade rapidly under simulated mammalian digestive conditions. An acute mouse gavage study confirmed that the NPTII protein caused no deleterious effects when administered by gavage at a cumulative target dosage of up to 5000 mg/kg of body weight. This dosage correlates to at least a million fold safety factor relative to the average daily consumption of potato or tomato, assuming all the potatoes or tomatoes consumed contained the NPTII protein. These results, along with previously published information, confirm that ingestion of genetically engineered plants expressing the NPTII protein poses no safety concerns.

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References

  1. Gasser, C.S. and Fraley, R.T. 1989. Genetically engineering plants tor crop improvement. Science 244: 1293–1299.

    Article  CAS  PubMed  Google Scholar 

  2. Casper, R. and Landsman, J. 1992. Summary of results, p. 12–14. In: The Biosafety Results Of Field Tests Of Genetically Modified Plants And Microorganisms. Casper, R. and Landsmann, J., Biologische Bundesanstalt fur Land- und Forstwirtschaft, Braunschweig, Germany.

    Google Scholar 

  3. Animal and Plant Health Inspection Service. 1992. Proposed interpretive ruling in connection with Calgene, Inc. petition for determination of regulatory status of Flavr Savr® Tomato. Fed. Reg. 57: 31170.

  4. Calgene, Inc. 1990. Request for advisory opinion KanR gene: safety and use in the production of genetically engineered plants. FDA Docket Number: 90A-0416.

  5. Calgene, Inc. 1993. Food additive petition for the APH(3′)II as a processing aid. FDA docket number: 93F-0232.

  6. Nap, J.P., Bijvoet, J. and Stikema, W.J. 1992. Biosafety of kanamycin-resistant transgenic plants: an overview. Transgenic Crops 1: 239–249.

    Article  CAS  Google Scholar 

  7. Flavell, R.B., Dart, E., Fuchs, R.L. and Fraley, R.T. 1992. Selectable marker genes: safe for plants? Bio/Technology 10: 141–144.

    CAS  Google Scholar 

  8. Guyton, A.C. 1981. Digestion and absorption in the gastointestinal tract, p. 816–826. In Textbook of Medicinal Physiology, 6th Ed. W.B. Saunders Co., Philadelphia.

    Google Scholar 

  9. Nielson, S.S. 1988. Degradation of bean proteins by endogenous and exogenous proteases—a review. Cereal Chem. 65: 435–442.

    Google Scholar 

  10. Marquez, U.M.L. and Lajolo, F.M. 1981. Composition and digestibility of albumin, globulins, and glutelins from Phaseolus vulgaris. J. Agric. Food Chem. 29: 1068–1074.

    Article  CAS  PubMed  Google Scholar 

  11. Zikakis, J.P., Rzucidlo, S.J. and Biasotto, N.O. 1977. Persistence of bovine milk xanthine oxidase activity. J. Dairy Science 60: 533–541.

    Article  CAS  Google Scholar 

  12. Tilch, C. and Elias, P.S. 1984. Investigation of the mutagenicity of ethylphenyl-glycidate. Mutation Research 138: 1–8.

    Article  CAS  PubMed  Google Scholar 

  13. Akeson, W.R. and Stahmann, M.A. 1964. A pepsin pancreatin digest index of protein quality evaluation. J. Nutrition 83: 257–261.

    Article  CAS  Google Scholar 

  14. Fuller, M.F. (Ed.). 1991. In Vitro Digestion for Pigs and Poultry. C.A.B. International, Wallingford, UK.

    Google Scholar 

  15. Alam, A.S., Hagerman, L.M. and Imondi, A.R. 1980. Bioavailability of sulpiride tablet and capsule in dogs. Arch. Int. Pharmodyn. Ther. 247: 180–189.

    CAS  Google Scholar 

  16. Doherty, A.M., Kaltenbronn, J.S., Hudspeth, J.P., Repine, J.T., Roark, W.H., Sircar, I., Tinney, F.J., Connolly, C.J., Hodges, J.C., Taylor, M.D., Batley, B.L., Ryan, M.J., Essenburg, A.D., Rapundalo, S.T., Weishaar, R.E., Humblet, C. and Lumney, E.A. 1991. New inhibitors of human renin that contain novel replacements at the P2 site. J. Med. Chem. 34: 1258–1271.

    Article  CAS  PubMed  Google Scholar 

  17. Li, A.P. 1989. Protein toxicology applied to biotechnology products. In: Genetically Engineered Plants: Scientific Issues In Their Regulation For Animal Feed And Human Food Uses, Workshop at Boyce Thompson Institute for Plant Research and Cornell University, Ithaca, New York.

    Google Scholar 

  18. Jones, D.D. and Maryanski, J.H. 1991. Safety considerations in the evaluation of transgenic plants for human food, p. 64–82. In: Risk Assessment In Genetically Engineering: Environmental Release Of Organisms. Levin, M. and Strauss, H. (Eds.). McGraw-Hill, New York.

    Google Scholar 

  19. Pariza, M.W. and Foster, E.M. 1983. Determining the safety of enzymes used in food processing. J. Food Protection 46: 453–468.

    Article  CAS  Google Scholar 

  20. Pesticide testing guidelines. Subdivision M. Microbial and Biochemical Pest Control Agents. PB89-211676. National Technical Information Service, Springfield, VA.

  21. Fuchs, R.L., Heeren, R.A., Gustafson, M.E., Rogan, G.J., Bartnicki, D.E., Leimgruber, R.M., Finn, R.F., Hershman, A. and Berberich, S.A. 1993. Purification and characterization of microbially expressed neomycin phospho-transferase II (NPTII) protein and its equivalence to the plant expressed protein. Bio/Technology 11: This issue.

  22. Kessler, D.A., Taylor, M.R., Maryanski, J.H., Flamm, E.L. and Kahl, L.S. 1992. The safety of foods developed by biotechnology. Science 256: 1747–1749.

    Article  CAS  PubMed  Google Scholar 

  23. Taylor, S.L., Nordlee, J.A. and Bush, R.K. 1992. Food allergies, p. 316–329. In: Food Safety Assessment, Ginley, J.W., Robinson, S.F. and Armstrong, D.J. (Eds.). ACS Symposium Series 484. ACS, Washington, DC.

    Chapter  Google Scholar 

  24. Taylor, S.L., Lemanske, R.F. Jr, Bush, R.K. and Busse, W.W. 1987. Food allergens: structure and immunologic properties. Ann. Allergy 59: 93–99.

    CAS  PubMed  Google Scholar 

  25. Sleisenger, M.H. and Fordtran, M.D. 1989. Gastrointestinal Disease, Volume 1. In: Pathophysiology Diagnosis Management, 4th Ed. W. B. Saunders Company, Philadelphia.

    Google Scholar 

  26. Davenport, H.W. 1971. Physiology of the Digestive Tract, 3rd Ed. Year Book Medical Publishers, Inc., Chicago.

    Google Scholar 

  27. Fuchs, R.L., Stone, T.B. and Lavrik, P.B. 1993. Risk assessment: a technical perspective. In: NABC Report 5, Agricultural Biotechnology: A Public Conversation About Risk, J. Fessenden MacDonald (Ed.). National Agricultural Biotechnology Council. Ithaca, NY.

    Google Scholar 

  28. United States Enironmental Protection Agency (USEPA). 1984. Dietary residue evaluation system (ORES) (formerly TAS: Tolerance assessment system). Washington, D.C.

  29. Food and Drug Administration, Department of Health and Human Services. 1992. Statement of Policy: Foods Derived from New Plant Varieties. Fed. Reg. 57: 22984–23005.

  30. International Food Biotechnology Council. 1990. Biotechnologies and food: assuring the safety of foods produced by genetic modification. Reg. Toxicol. Pharmacol. 12: S1–S196.

  31. The United States Pharmacopeia, Vol. XXII, p. 1788–1789. United States Phar-macopeial Convention, Inc. Rockville, MD.

  32. Ryle, A.P. 1984. Pepsins, gastricsins and their zymogens, p. 223–238. In: Methods of Enzymatic Analysis, Vol. V. Bergmeyer, H.U. (Ed.). Verlag Chemie, Weinheim.

    Google Scholar 

  33. Laemmli, U.K. 1970. Cleavage of structural proteins during assembly of the head of the bacteriophage T4. Nature 227: 680–685.

    Article  CAS  PubMed  Google Scholar 

  34. Matsudaira, P. 1987. Sequence from picomole quantities of proteins electro-blotted onto polyvinylidene difluoride membranes. J. Biol. Chem. 262: 10035–10038.

    CAS  PubMed  Google Scholar 

  35. Goldman, P.R. and Northrop, D.B. 1976. Purification and spectrophotometric assay of neomycin phosphotransferase II. Biochem. Biophys. Res. Comm. 69: 230–236.

    Article  CAS  PubMed  Google Scholar 

  36. Dunnett, C.W. 1955. A multiple comparison procedure for comparing several treatments with a control. J. Am. Stat. Assoc. 50: 10967–1121.

    Article  Google Scholar 

  37. Dixon, W.J. and Massey, F.J. Jr 1969. Introduction to Statistical Analysis, 3rd Ed. McGraw-Hill Co., NY.

    Google Scholar 

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  1. Roy L. Fuchs: Corresponding author.

Authors and Affiliations

  1. The Agricultural Group, A Unit of Monsanto Company, 700 Chesterfield Village Parkway, Chesterfield, MO, 63198

    Roy L. Fuchs, Joel E. Ream, Bruce G. Hammond, Mark W. Naylor, Richard M. Leimgruber & Sharon A. Berberich

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  2. Joel E. Ream
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Fuchs, R., Ream, J., Hammond, B. et al. Safety Assessment of the Neomycin Phosphotransferase II (NPTII) Protein. Nat Biotechnol 11, 1543–1547 (1993). https://doi.org/10.1038/nbt1293-1543

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