Safety assessment of a modified acetolactate synthase protein (GM-HRA) used as a selectable marker in genetically modified soybeans.
Acetolactate synthase (ALS) enzymes have been isolated from numerous organisms including soybeans (Glycine max; GM-ALS) and catalyze the first common step in biosynthesis of branched chain amino acids. Expression of an ALS protein (GM-HRA) with two amino acid changes relative to native GM-ALS protein in genetically modified soybeans confers tolerance to herbicidal active ingredients and can be used as a selectable transformation marker. The safety assessment of the GM-HRA protein is discussed. Bioinformatics comparison of the amino acid sequence did not identify similarities to known allergenic or toxic proteins. In vitro studies demonstrated rapid degradation in simulated gastric fluid (<30s) and intestinal fluid (<1min). The enzymatic activity was completely inactivated at 50 degrees C for 15 min demonstrating heat lability. The protein expressed in planta is not glycosylated and genetically modified soybeans expressing the GM-HRA protein produced similar protein/allergen profiles as its non-transgenic parental isoline. No adverse effects were observed in mice following acute oral exposure at a dose of at least 436 mg/kg of body weight or in a 28-day repeated dose dietary toxicity study at doses up to 1247 mg/kg of body weight/day. The results demonstrate GM-HRA protein safety when used in agricultural biotechnology.
Mathesius
C A
CA
Pioneer Hi-Bred International, Inc., Ankeny, IA, USA.
Barnett
J F
JF
Cressman
R F
RF
Ding
J
J
Carpenter
C
C
Ladics
G S
GS
Schmidt
J
J
Layton
R J
RJ
Zhang
J X Q
JX
Appenzeller
L M
LM
Carlson
G
G
Ballou
S
S
Delaney
B
B
D000095:Acetolactate Synthase / Q000008:administration & dosage / Q000302:isolation & purification / Q000633:toxicity*; D000383:Agriculture / Q000379:methods; D000595:Amino Acid Sequence; D000818:Animals; D001709:Biotechnology / Q000379:methods; D019295:Computational Biology / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004795:Enzyme Stability; D005260:Female; D030841:Food, Genetically Modified / Q000633:toxicity*; D005750:Gastric Juice / Q000378:metabolism; D053209:Herbicide Resistance; D006358:Hot Temperature; D007419:Intestinal Secretions / Q000378:metabolism; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D030821:Plants, Genetically Modified / Q000201:enzymology*; D013025:Glycine max / Q000201:enzymology* / Q000235:genetics; D018675:Toxicity Tests
D000095:Acetolactate Synthase