Evaluation of Protein Safety in the Context of Agricultural Biotechnology

Delaney B, Astwood JD, Cunny H, Conn RE, Herouet-Guicheney C, MacIntosh S, Meyer LS, Privalle L, Gao Y, Mattsson J, Levine M
Food and Chemical Toxicology
January 1, 2008

Download

Task Force #6

Food and Chemical Toxicology. 2008;46:S71-S97

Published as a supplement in Food and Chemical Toxicology, this unique document brings together a considerable body of information about the science underlying protein safety assessment and puts forth a scientifically based two-tiered, weight-of evidence strategy to assess the safety of novel proteins used in the context of agricultural biotechnology.

Recommendations draw upon knowledge of the biological and chemical characteristics of proteins and testing methods for evaluating potential intrinsic hazards of chemicals. The application of this strategy is presented using examples of transgenic proteins applied for agricultural input and output traits in genetically modified crops along with recommendations for future research considerations related to protein safety assessment.

To download this article, click here.

References

Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P., 2002. Biology of the Cell. Garland Science, New York, NY.
Alouf, J., Freer, J., 1999. The Comprehensive Sourcebook of Bacterial Protein Toxins, second ed. Academic Press, New York, NY.
Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., 1990. Basic local alignment search tool. J. Mol. Biol. 215, 403–410. LINK
Astwood, E.G., 1970. Anterior pituitary hormones and related substances. In: Goodman, L.S., Gilman, A. (Eds.), The Pharmacological Basis of Therapeutics, fourth ed. MacMillan Publishing Company, New York, pp. 1538–1565.
Astwood, J.D., Leach, J.N., Fuchs, R.L., 1996. Stability of food allergens to digestion in vitro. Nature Biotech. 14, 1269–1273. LINK
Atherton, K., 2002. Safety assessment of genetically modiﬁed crops. Toxicology, 421–426. LINK
Barlow, S.M., Kozianski, G., Wurtzen, G., Schlatter, J., 2001. Threshold of toxicological concern for chemical substances in the diet. A report of a workshop held on 5–6 of October, 1999, in Paris, France organized by ILSI Europe Threshold of Toxicological Concern Task Force. Food Chem. Toxicol. 39, 893–905. LINK
Betz, F., Hammond, B.G., Fuchs, R.L., 2000. Safety and advantages of Bacillus thuringiensis-protected plants to control insect pests. Regul. Toxicol. Pharmacol. 32, 156–173. LINK
Beutler, B., Milsark, I.W., Cerami, A.C., 1985. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal eﬀects of endotoxin. Science 229, 869–871. LINK
Broekaert, W.F., Cammue, B.P.A., De Bolle, M.F.C., Thevissen, K., De Samblanx, G.W., Osborn, R.W., 1997. Antimicrobial peptides from plants. Crit. Rev. Plant Sci. 16, 297–323. LINK
Cellini, F., Chesson, A., Colquhoun, I., Constable, A., Davies, H.V., Engel, K.H., Gatehouse, A.M.R., Ka¨ renlampi, S., Kok, E.J., Leguay, J.-J., Lehesranta, S., Noteborn, H.P.J.M., Pedersen, J., Smith, M., 2004. Unintended eﬀects and their detection in genetically modiﬁed crops. Food Chem. Toxicol. 42, 1089–1125. LINK
Chassy, B., 2002. Food safety evaluation of crops produced through biotechnology. J. Am. College Nutr. 21, 1665–1735. LINK
Chassy, B., Hlywka, J.J., Kleter, G.A., Kok, E.J., Kuiper, H.A., McGloughlin, M., Munro, I.C., Phipps, R.H., Reid, J.E., 2004. Nutritional and safety assessments of foods and feeds nutritionally improved through biotechnology. Food Nutr. Bull. 26, 436–442. LINK
Cheeke, P.R., Shull, L.R., 1985. Proteins and amino acids. Natural Toxicants in Feed and Poisonous Plants. AVI Publishing Co., Westport CT, pp. 235–291.
Cheeseman, M.A., Machuga, E.J., Bailey, A.B., 1999. A tiered approach to threshold of regulation. Food Chem. Toxicol. 37, 387–412. LINK
Codex Alimentarius Commission, 2003. Alinorm 03/34: Joint FAO/ WHO Food Standard Programme, Codex Alimentarius Commission, Twenty-Fifth Session, Rome, Italy, 30 June–5 July,2003. Appendix III, Guideline for the conduct of food safety assessment of foods derived from recombinant-DNA plants and Appendix IV, Annex on the assessment of possible allergenicity, pp. 47– 60. LINK
Collins, V.L., Hackett, J., 1991. Sequence of the phosphomannose isomerase encoding gene of Salmonella typhimurium. Gene 103, 135– 136. LINK
Conolly, R.B., Lutz, W.K., 2004. Nonmonotonic dose–response relationships: mechanistic basis, kinetic modeling, and implications for risk assessment. Toxicol. Sci. 77, 151–157. LINK
Constable, A., Jonas, D., Cockburn, A., Davi, A., Edwards, G., Hepburn, P., Herouet-Guicheney, C., Knowles, B., Moseley, B., Oberdo¨ rfer, R., 2007. ‘History of safe use’ as applied to the safety assessment of novel foods and foods derived from genetically modiﬁed organisms. Food Chem. Toxicol. 45, 2513–2525. LINK
Crickmore, N., Zeigler, D.R., Feitelson, J., Schnepf, E., Van Rie, J., Lereclus, D., Baum, J., Dean, D.H., 1998. Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol. Mol. Biol. Rev. 62, 807–813. LINK
Darzins, A., Franz, B., Vanags, R.I., Chakrabarty, A.M., 1986. Nucleotide sequence analysis of the phosphomannose isomerase gene(pmi) of Psuedomaonas aeruginosa and comparison with the corresponding Escherichia coli gene manA. Gene 42, 293–302. LINK
Day, P.R., 1996. The biology of plant proteins. CRC Crit. Rev. Food Sci. Nutr. 36, 39–47. LINK
De Samblanx, G.W., Goderis, I.J., Thevissen, K., Raemaekers, R., Fant, F., Borremans, F., Acland, D.P., Osborn, R.W., Patel, S., Broekaert, W.F., 1997. Mutational analysis of a plant defensin from radish (Raphanus sativus L.) reveals two adjacent sites important for antifungal activity. J. Biol. Chem. 271, 1171–1179. LINK
Devlin, T.M., 2002. Textbook of Biochemistry with Clinical Correlations, ﬁfth ed. John Wiley & Sons, New York, NY.
Doolittle, R.F., 1990. Searching through sequence databases. Methods Enzymol. 183, 99–110. LINK
Duval, A., Malecot, C.O., Pelhate, M., Rochat, H., 1989. Changes in Na channel properties of frog and rat skeletal muscles induced by the AaH II toxin from the scorpion Androctonus australis. Pﬂugers Arch. 415, 361–371. LINK
Eckhoﬀ, S.R., Rausch, K.D., Fox, E.J., Tso, C.C., Wu, X., Pan, Z., Buriak, P., 1993. A laboratory wet milling procedure to increase reproducibility and accuracy of product yields. Cereal Chem. 70, 727–732. LINK
EFSA, 2006a. Guidance document of the Scientiﬁc Panel on Genetically Modiﬁed Organisms for the risk assessment of genetically modiﬁed plants and derived food and feed. EFSA J. 9, 1–100. LINK
EFSA, 2006b. Safety and Nutritional Assessment of GM Plant derived Foods/Feed: The role of animal feeding trials. Draft for public consultation. December 15, 2006. LINK
FAO/WHO, 1991. Strategies for assessing the safety of foods produced by biotechnology. Report of a joint FAO/WHO Expert Consultation. WHO, Geneva. LINK
FAO/WHO, 1996. Biotechnology and Food Safety Report. Report of a joint FAO/WHO Expert Consultation, Rome, Italy, 30 September–4 October, 1996. FAO Food and Nutrition Paper 61. Food and Agriculture Organisation of the United Nations, Rome. LINK
Franz, D.R., Jax, N.K., 1997. Ricin toxin. In: Zajtchuk, R., Bellamy, R.F. (Eds.), Textbook of Military Medicine: Medical aspects of chemical and biological warfare. Oﬃce of The Surgeon General Department of the Army, United States of America, TMM Publications, Washington, DC. LINK
Frawley, J.P., 1967. Scientiﬁc evidence and common sense as a basis for food-packaging regulations. Food Cosmet. Toxicol. 5, 293–308. LINK
Finklestein, R.A., 1973. Cholera. CRC Crit. Rev. Microbiol. 2, 553–623. LINK
Fricker, G., Drewe, J., 1996. Current concepts in intestinal peptide absorption. J. Pept. Sci. 2, 195–211. LINK
Fu, T.-J., Abbott, U.R., Hatzos, C., 2002. Digestibility of food allergens and nonallergenic proteins in simulated gastric ﬂuid and simulated intestinal ﬂuid – a comparative study. J. Agric. Food Chem. 50, 7154– 7160. LINK
Gatehouse, A.M., Norton, E., Davison, G.M., Babbe, S.M., Newell, C.A., Gatehouse, J.A., 1999. Digestive proteolytic activity in larvae of tomato moth, Lacanobia oleracea; eﬀects of plant protease inhibitors in vitro and in vivo. J. Insect Physiol. 45, 545–5558. LINK
Gerrior, D., Bente, L., Hiza, H., 2004. Nutrient content of the US food supply, 1909–2000 Home Economics Research Report No. 56. US Department of Agriculture, Center for Nutrition Policy and Promotion. LINK
Gibson, J., 2006. Bioinformatics ofprotein allergenicity. Mol. Nutr. Food Res. 50 (7), 591–670. LINK
Gill, M.D., 1982. Bacterial toxins: a table of lethal amounts. Microbiol. Rev. 46, 86–94. LINK
Gill, M.D., 1987a. Bacterial toxins: description. In: Laskin, A.I., Lechevalier, H.A. (Eds.), CRC Handbook of Microbiology, second ed., . In: Toxins and Enzymes, vol. VIII CRC Press, Boca Raton, FL, pp. 3–18.
Gill, M.D., 1987b. Bacterial toxins: lethal amounts. In: Laskin, A.I., Lechevalier, H.A. (Eds.), CRC Handbook of Microbiology, second ed., . In: Toxins and Enzymes, vol. VIII CRC Press, Boca Raton, FL, pp. 127–135.
Goldsworthy, A., Street, H.E., 1965. The carbohydrate nutrition of tomato roots VIII. The mechanism of the inhibition of D-mannose of the respiration of excised roots. Ann. Bot. 29, 45–58. LINK
Goodman, R.E., Heﬂe, S.L., Taylor, S.L., van Ree, R., 2005. Assessing genetically modiﬁed crops to minimize the risk of increased food allergy: a review. Int. Arch. Allergy Immunol. 137, 153–166. LINK
Harrison, L.A., Bailey, M.R., Naylor, M., Ream, J., Hammond, B., Nida, D.L., Burnette, B., Nickson, T.E., Mitsky, T., Taylor, M.L., Fuchs, R.L., Padgette, S.R., 1996. The expressed protein in glyphosate tolerance soybean, 5-enolpryruvyl-shikimate-3-phosphate synthase from Agrobacteriuin sp. strain CP4, is rapidly digested in vitro and is not toxic to acutely gavaged mice. J. Nutr. 126, 728–740. LINK
Hauschild, A., 1989. Clostridium botulinum. In: Doyle, M.P. (Ed.), Foodborne Bacterial Pathogens. Marcel Dekker, New York, NY, pp. 111–190.
Heﬂe, S.L., Nordlee, J.A., Taylor, S.L., 1996. Allergenic foods. CRC Crit. Rev. Food Sci. Nutr. 36, 69–89.
Herman, R.A., Schafer, B.W., Korjagin, V.A., Ernest, A.D., 2003. Rapid digestion of Cry34Ab1 and Cry35Ab1 in simulated gastric ﬂuid. J. Agric. Food Chem. 51, 6823–6827. LINK
Herman, R.A., Storer, N.P., Gao, Y., 2006. Digestion assays in allergenicity assessment of transgenic proteins. Environ. Health Persp. 114, 1154–1157. LINK
Hofte, H., Whitely, H.R., 1989. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol. Rev. 53, 242–255. LINK
Herouet, C., Esdaile, D., Mallyon, B.A., Debruyne, E., Schulz, A., Hendrickx, K., van der Klis, R.-J., Rouan, D., 2005. Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate–ammonium herbicide in transgenic plants. Regul. Toxicol. Pharmacol. 41, 134–149. LINK
Hileman, R.E., Silvanovich, A., Goodman, R.E., Rice, E.A., Holleschak, G., Astwood, J.D., Heﬂe, SL., 2002. Bioinformatic methods for allergenicity assessment using a comprehensive allergen database. Int. Arch. Allergy Immunol. 128, 280–291. LINK
ISAAA (International Service for the acquisition of agri-biotech applications), 2006. <http://www.isaaa.org/>.
Ishiguro, M., Mitarai, M., Harada, H., Sekine, I., Kikutani, M., 1983. Biochemical studies on oral toxicity of ricin. I. Ricin administered orally can impair sugar absorption by the rat small intestine. Chem. Pharm. Bull. 31, 3222–3227. LINK
Kaneda, M., Oyama, Y., Ikemoto, Y., Akaike, N., 1989. Scorpion toxin prolongs an inactivation phase of the voltage-dependent sodium current in rat isolated single hippocampal neurons. Brain Res. 487, 192–195. LINK
Kroes, R., Galli, C., Munro, I., Schilter, B., Tran, L.-A., Walker, R., Wurtzen, G., 2000. Threshold of toxicological concern for chemical substances in the diet: a practical tool for assessingthe need for toxicity testing. Food Chem. Toxicol. 38, 255–312. LINK
Kroes, R., Renwick, A.G., Cheeseman, M., Kleiner, J., Mangelsdorﬀ, I., Piersma, A., Schilter, B., Schlatter, J., van Schthorst, F., Vos, J.G., Wurtzen, G., 2004. Structure-based thresholds of toxicological concern (TCC): guidance for applications to substances present at low levels in the diet. Food Chem. Toxicol. 42, 65–83. LINK
Kuiper, H.A., Kleter, G.A., Noteborn, H.P.J.M., Kok, E.J., 2002. Substantial equivalence – an appropriate paradigm for the safety assessment of genetically modiﬁed foods. Toxicology, 427–431. LINK
Kutzner, H.J., 1981. The family Streptomycetaceae. In: Starr, M.P., Stolp, H., Tru¨ per, H.G., Ballows, A., Schlegel, H.G. (Eds.), The Prokaryotes: A Handbook on Habitats, Isolation and Identiﬁcation of Bacteria. Springer-Verlag, Berlin, pp. 2028–2072.
Ladics, G.S., Bannon, G.A., Silvanovich, A., Cressman, RF., 2007. Comparison of conventional FASTA identity searches with the 80 amino acid sliding window FASTA search for the elucidation of potential identities to known allergens. Mol. Nutr. Food Res. 51, 985–998. LINK
Lee, B.T., Matheson, N.K., 1984. Phosphomanoseisomerase and phos-phoglucoisomerase in seeds of Cassia coluteoides and some other legumes that synthesize galactomannan. Phytochemistry 23, 983–987. LINK
Lee, D., Gurevitz, M., Adams, M.E., 2000. Modiﬁcation of synaptic transmission and sodium channel inactivation by the insect-selective scorpion toxin LqhaIT. J. Neurophysiol. 83, 1181–1187. LINK
Liener, I.E., 1994a. Antinutritional factors related to proteins and amino acids. In: Hui, Y.H., Gorham, J.R., Murell, K.D., Cliver, D.O. (Eds.), Foodborne Disease Handbook. Disease Caused by Hazardous Substances. Marcell Dekker, New York, NY, pp. 261–309.
Leiner, I.E., 1994b. Implications of antinutritional components in soybean foods. CRC Crit. Rev. Food Sci. Nutr. 34, 31–67. LINK
Lyerly, D.M., Saum, K.E., MacDonald, D.K., Wilkins, T.D., 1985. Eﬀects of Clostridium diﬃcile toxins given intragastrically to animals. Infect Immun. 47, 349–352. LINK
Magalhaes, M.D.M., Vina, G., Arantes, R.M.E., Santos, T.M., Cunha-Melo, J.R., 1998. The mouse as an experimental model for Tityus serrulatus scorpion envenoming. Acta Cir. Bras. 13, 260–263. LINK
Maksymowych, A.B., Reinhard, M., Malizio, C.J., Goodnough, M.C., Johnson, E.A., Simpson, L.L., 1999. Pure botulinum neurotoxin is absorbed from the stomach and small intestine and produces peripheral neuromuscular block. Infect Immun. 67, 4708–4712. LINK
Metcalfe, D.D., Astwood, J.D., Townsend, R., Sampson, H.A., Taylor, S.L., Fuchs, R.L., 1996. Assessment of the allergenic potential of foods from genetically engineered crop plants. CRC Crit. Rev. Food Sci. Nutr. 36, 165–186. LINK
Miles, S.J., Guest, J.R., 1984. Nucleotide sequence and transcriptional start point of the phosphomannose isomerase gene (manA)of Escherichia coli. Gene 32, 41–48. LINK
Morgan, D.R., Street, H.E., 1959. The carbohydrate nutrition of tomato roots. VII. Sugars, sugar phosphates and sugar alcohols as respiratory substrates for excised roots. Ann. Bot. 23, 89–105. LINK
Munro, I.C., Kennepohl, E., Kroes, R., 1999. A procedure for the safety evaluation of ﬂavouring substances. Food Chem. Toxicol. 37, 207–232. LINK
Nair, R.S., Fuchs, R.L., Schuette, S.A., 2002. Current methods for assessingsafety of genetically modiﬁed crops as exempliﬁed by data on Roundup Ready soybeans. Toxicol. Pathol. 30, 117–125. LINK
Nida, D.L., Kolacz, K.H., Buehler, R.E., Deaton, W.R., Schuler, W.R., Armstrong, T.A., Taylor, M.L., Ebert, C.C., Rogan, G.J., 1996. Glyphosate-tolerant cotton: genetic characterization and protein expression. J. Agric. Food Chem. 44, 1960–1966. LINK
Niehues, R., Hasilik, M., Alton, G., Korner, C., Schiebe-Sukumar, M., Koch, H.G., Zimmer, K., Wu, R., Harms, E., Reiter, K., von Figura, K., Freeze, H.H., Harms, H.K., Marquardt, T., 1998. Carbohydrate-deﬁcient glycoprotein syndrome type 1b. J. Clin. Invest. 101, 1414– 1420. LINK
OECD, 2002. Guideline for the testing of chemicals: acute oral toxicity. OECD Test Guideline 401. Organization for Economic Co-operation and Development, Paris, France. LINK
OECD, 1995. Guideline for the testing of chemicals: repeated dose 28-day oral toxicity study in rodents. OECD Test Guideline 407. Organization for Economic Co-operation and Development, Paris, France. LINK
PFAM, 2005 Protein families database of alignments and HMMs. LINK
Padgette, S.R., Kolack, K.H., Delannay, X., Re, D.B., La Vallee, B.J., Tinius, C.N., Rhodes, W.K., Otero, Y.I., Barry, G.F., Eichholtz, D.A., Peschke, Nida, D.L., Taylor, N.B., Kishore, G.M., 1995. Development, identiﬁcation, and characterization of a glyphosate-tolerant soybean line. Crop Sci. 35, 1451–1461. LINK
Padgette, S.R., Re, D.B., Gasser, C.S., Eichholtz, D.A., Frazier, R.B., Hironaka, C.M., Levine, E.B., Shah, D.M., Fraley, R.T., Kishore, G.M., 1991. Site-directed mutagenesis of a conserved region of the 5-enolpyruvylshikimate-3-phosphate synthase active site. J. Biol. Chem. 266, 22364–22369. LINK
Padgette, S.R., Re, D.B., Barry, G.F., Eichholtz, D.E., Delannay, F.X., Fuchs, R.L., Kishore, G.M., Fraley, R.T., 1996. New weed control opportunities: development of soybeans with a roundup ready gene. In: Duke, S.O. (Ed.), Herbicide-resistant Crops: Agricultural, Environmental, Economic, Regulatory, and Technical Aspects. CRC Press, New York, NY, pp. 53–84.
Pearson, W.R., Lipman, D.J., 1988. Improved tools for biological sequence comparison. Proc. Natl. Acad. Sci. USA 85, 2440–2448. LINK
Popoﬀ, M.R., 1998. Interactions between bacterial toxins and intestinal cells. Toxiconomy 36, 665–685. LINK
Proudfoot, A.E.I., Turcatti, G., Wells, T.N.C., Payton, M.A., Smith, D.J., 1994a. Puriﬁcation, cDNA cloning and heterologous expression of human phosphomannose isomerase. Eur. J. Biochem. 219, 415–423. LINK
Proudfoot, A.E., Payton, M.A., Wells, T.N., 1994b. Puriﬁcation and characterization of fungal and mammalian phosphomannose isomerases. J. Protein. Chem. 13, 619–627. LINK
Powell-Abel, P., Nelson, R.S., De, B., Hoﬀmann, N., Rogers, S.G., Fraley, R.T., Beachy, R.N., 1980. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232, 738–743. LINK
Ratner, B., Untracht, S., Collins-Williams, C., 1952. Allergenicity of modiﬁed and processed foodstuﬀs. 1. The use of a dual ingestion passive transfer test to determine the allergenicity of foodstuﬀs in man. Ann. Allergy 10, 675–681. LINK
Reed, J., Privalle, L., Powell, M.L., Meghji, M., Dawson, J., Dunder, E., Suttie, J., Wenk, A., Launis, K., Kramer, C., Chang, Y., Hansen, G., Wright, M., 2001. Phosphomannose isomerase: an eﬃcient selectable marker for plant transformation. In vitro cell. Dev. Biol. – Plant 37, 127–132. LINK
Roberts, P.R., Burney, J.D., Black, K.W., Zaloga, G.P., 1996. Eﬀect of chain length on absorption of biologically active peptides from the gastrointestinal tract. Digestion 60, 332–337. LINK
Robertus, J.D., 1991. The structure and action of ricin, a cytotoxic N-glycosidase. Semin. Cell Biol. 2, 23–30. LINK
Sakaguchi, G., Ohishi, T., Kozaki, S., 1988. Botulism – structure and chemistry of botulinum toxins. In: Herdegree, M.C., Tu, A.T. (Eds.), Handbook of Natural Toxins. Marcel Dekker, New York, NY, pp. 191–216.
Sanders, P.R., Lee, T.C., Groth, M.E., Astwood, J.D., Fuchs, R.L., 1998. Safety assessment of insect-protected corn. In: Thomas, J.A. (Ed.), Biotechnology and Safety Assessment, second ed. Taylor and Francis, Oxford, UK, pp. 241–256.
Schmidt, M., Arnold, W., Niemann, A., Kleickmann, A., Puhler, A., 1992. The Rhizobium meliloti pmi gene encodes a new type of phosphomannose isomerase. Gene 122, 35–43. LINK
Shinabarger, D., Berry, A., May, T.B., Rothmel, R., Fialho, A., Chakrabarty, A.M., 1991. Puriﬁcation and characterization of phosphomannose isomerase-guanosine diphospho-D-mannose pyrophosphorylase.A bifunctional enzyme in the alginate biosynthetic pathway of Pseudomonas aeruginosa. J. Biol. Chem. 266, 2080–2088. LINK
Sidell, F.R., Takafuji, E.T., Franz, D.R., 1997. Part I: Warfare, weaponry, and the casualty. In: Sidell, F.R., Takafuji, E.T., Franz, D.R. (Eds.), Textbook of Military Medicine, . In: Medical Aspects of Chemical and Biological Warfare, vol. 3. Borden Institute, Walter Reed Medical Center, Washington, DC.
Smith, D.J., Proudfoot, A.E.I., Freidli, L., Klig, L.S., Paravicini, G., Payton, M.A., 1992. PMI40, an intron-containing gene required for early steps in yeast mannosylation. Mol. Cell. Biol. 12, 2924– 2930. LINK
Skjelkvale, R., Uemura, T., 1977. Experimental diarrhoea in human volunteers following oral administration of Clostridium perfringens enterotoxin. J. Appl. Bacteriol. 43, 281–286. LINK
Sjoblad, R.D., McClintock, J.T., Engler, R., 1992. Toxicological consid-erations for protein components of biological pesticide products. Regul. Toxicol. Pharmacol. 15, 3–9. LINK
Smolin, L.A., Grosvenor, M.B., 2000. Nutrition Science & Applications, third ed. Saunders College Publishing, New York, NY.
Stallings, W.C., Abdel-Meguid, S.S., Lim, L.W., Shieh, H.S., Dayringer, H.E., Leimgruber, N.K., Stegeman, R.A., Anderson, K.S., Sikorski, J.A., Padgette, S.R., Kishore, G.M., 1991. Structure and topological symmetry of the glyphosate target 5-enolpyruvylshikimate-3-phosphate synthase: a distinctive protein fold. Proc. Natl. Acad. Sci. USA 88, 5046–5050. LINK
Stelid, G., 1954. Toxic eﬀects of D-mannose, 2-desoxy-D-glucose, and D-glucosamine upon respiration and ion absorption in wheat roots. Physiol. Plant 7, 173–181. LINK
Strobel, S., 1998. Oral tolerance: immune responses to food antigens. In: Metcalf, D.D., Sampson, H., Simon, R. (Eds.), Food Allergy: Adverse Reactions to Foods and Food Additives. Blackwell Science Publishing, Massachusetts, pp. 107–136.
Suzuki, K., Tateda, K., Matsumoto, T., Gondaira, F., Tsujimoto, S., Yamaguchi, K., 2000. Eﬀects of interaction between Escherichia coli verotoxin and lipopolysaccharide on cytokine induction and lethality in mice. J. Med. Microbiol. 49, 905–910. LINK
Terras, F.R., Schoofs, H.M., De Bolle, M.F.C., Van Leuven, F., Rees, S.B., Vanderleyden, J., Cammue, B.P.A., Broekaert, W.F., 1992. Analysis of two novel classes of plant antifungal proteins from radish (Raphanus sativus L.) seeds. J. Biol. Chem. 267, 15301–15309. LINK
Thevissen, K., Ghazi, A., De Samblanx, G.W., Brownlee, C., Osborn, R.W., Broekaert, W.F., 1996. Fungal membrane responses induced by plant defensins and thionins. J. Biol. Chem. 271, 15018–15025. LINK
Thomas, K., Aalbers, M., Bannon, G.A., Bartels, M., Dearman, R.J., Esdaile, D.J., Fu, T.J., Glatt, C.M., Hadﬁeld, N., Hatzos, C., Heﬂe, S.L., Heylings, J.R., Goodman, R.E., Henry, B., Herouet, C., Holsapple, M., Ladics, G.S., Landry, T.D., MacIntosh, S.C., Rice, E.A., Privalle, L.S., Steiner, H.Y., Teshima, R., van Ree, R., Woolhiser, M., Zawodny, J., 2004. A multi-laboratory evaluation of a commonin vitro pepsin digestion assay protocol used in assessing the safety of novel proteins. Regul. Toxicol. Pharmacol. 39, 87–98. LINK
Thomas, K., Bannon, G., Heﬂe, S., Herouet, C., Holsapple, M., Ladics, G., MacIntosh, S., Privalle, L., 2005. In silico methods for evaluating human allergenicity to novel proteins: Bioinformatics Workshop Meeting Report, 23–24 February 2005. Toxicol. Sci. 88, 307–310. LINK
Thomas, K., Herouet-Guicheney, C., Ladics, G., Bannon, G., Cockburn, A., Crevel, R., Fitzpatrick, J., Mills, C., Privalle, L., Vieths, S., 2007. Evaluating the eﬀects of food processing on the potential human allergenicity of novel proteins: international workshop report. Food Chem. Toxicol. 45, 1116–1122. LINK
Thompson, C.J., Movva, N.R., Tizard, R., Crameri, R., Davies, J.E., Lauwereys, M., Botterman, J., 1987. Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus. EMBO J.6, 2519–2523. LINK
Tsume, Y., Taki, Y., Sakane, T., Nadai, T., Sezaki, H., Watabe, K., Kohno, T., Yamashita, S., 1996. Quantitative evaluation of the gastrointestinal absorption of protein into the blood and lymph circulation. Biol. Pharm. Bull. 19, 1332–1337. LINK
Uniprot-Swissprot Consortium, 2007. The Universal Protein Resource (UniProt). Nucleic Acids Research, vol. 35, Database issue D193– D197. LINK
US FDA, 1983. Toxicological principles for the safety assessment of direct food additives and color additives used in food. Redbook. US FDA, Bureau of Foods, Washington DC.
US EPA, 1998. Health effects test guidelines. OPPTS 870.1100 Acute oral toxicity. LINK
US EPA, 1996. EPA Fact Sheet for Bacillus thuringiensis subspecies kurstaki Cry 1A (b) Delta Endotoxin and Its Controlling Sequences as Expressed in Corn. “LINK
US EPA, 2000. Background document for the FIFRA scientiﬁc advisory panel on mammalian toxicity assessment guidance for protein plant pesticides. LINK
USP, 2000. The United States Pharmacopeia 24. Simulated gastric ﬂuid, TS. In: Board of Trustees (Eds.), The National Formulary 19. United States Pharmacopeial Convention, Inc., Rockville, MD, pp. 2235.
Van Haver, E., DeSchrijver, A., Devos, Y., Lievens, S., Renckens, S., Moens, W., 2003. Guidance notes for the safety assessment of genetically modiﬁed crops for food and feed use. Report of the Belgium Biosafety Advisory Council, Scientiﬁc Institute of Public Health. Royal Library of Belgium Deposit No. D/2003/2505/16.
Van Rie, J., Jansens, S., Hofte, H., Degheele, D., Van Mellaert, H., 1989. Speciﬁcity of Bacillus thuringiensis o-endotoxins. Eur. J. Biochem. 186, 239–247. LINK
Wang, G.K., Strichartz, G., 1982. Simultaneous modiﬁcations of sodium channel gating by two scorpion toxins. Biophys. J. 40, 175–179. LINK
Webb, K.E., 1990. Intestinal absorption of protein hydrolysis products: a review. J. Anim. Sci. 68, 3011–3022. LINK
Wehrmann, A., Van Vliet, A., Opsomer, C., Botterman, J., Schul, A., 1996. The similarities of bar and pat gene products make them equally applicable for plant engineers. Nature Biotechnol. 14, 1274–1278. LINK
Zhuang, M., Gill, S.S., 2002. Mode of action of Bacillus thuringiensis toxins. In: Voss, G., Ramos, G. (Eds.), Chemistry of Crop Protection, . In: Progress and Prospects in Science and Regulation. Wiley & Sons, New York, NY, pp. 213–236.