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Deteksi Keamanan Pangan
Transgenik
Dahrul Syah
Teknologi Pangan - IPB
Agenda
Pengantar
Substantial Equivalen
Beyond Substantial Equivalen
Pro Kontra
Pengaturan di Indonesia
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Biotech Products in
Food Supply
1. Fermentation products Yeasts and molds used to make cheese,
bread, wine, beer, soy sauce
2. Enzymes Lactase tablets to breakdown lactose in
milk - for allergies
Chymosin (rennet) for cheese
manufacture
Amylase in beer
3. Hormones
Bovine growth hormone(rbst)
Vitamin D
added milk
Insulin
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4. Whole foods/crops
FLAVR SAVR,Endless Summertomatoes
Virus resistantsquash
Insect resistantpotatoes, corn
Herbicide resistantsoybeans, canola
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Acceptance of Technologies
Public accepts new technologies with personal
benefits
rInsulin vs rBST
Benefits of many
bioengineered
products unknown or not
perceived as personal
Platform of omics
PLANT
TISSUE
DNA/mRNAs
PROTEINS
COMPONENTS
DIFFERENCES
AGRONOMICS
PHENOTYPICS
GENOMICS
PROTEOMICS
METABOLOMICS
STATISTICSUnintended effects?
Metabolite
expression
Protein
expression
Gene
expressionFunctionStructureData
integration
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Potential Food Safety Effects
Toxicants
New Substances
Nutrients
Allergenicity
Other Effects - Unintended
-- IFT Expert Report on Biotechnology and Foods, 2000
Considerations in Safety
Evaluations Substantial Equivalence based on
comparison of recombinant product with
traditional product
Not an absolute guarantee of safety
Process to establish that no NEW hazards have
been introduced into the plant or product
Toxins?
Nutrient Effects? Proteins, fats, carbohydrates,
vitamins, minerals
Allergens?
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Genetic characterisation
Comparison
with Traditional
Counterpart
Agronomic characterisation
Chemical characterisationnutrientstoxicants
allergensDietary patterns
History of safe useSelection of Traditional
Counterpart
Equivalency testing
Nutrients: Carbohydrates
Proteins
Lipids
Vitamins
Minerals
Allergens
Asam Lemak
Asam Amino
Kesetaraan Substansial
Tempe yang dibuat dari kedele import dan kedele lokal
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Oil Oil
1.EquivalentNo further testing
2.Equivalence except forpredictable, well-defined traitsFocused evaluation on limiteddifferences
3.Lack of equivalence, multiplecomplex differences or notraditional counterpartRigorous case-by-case safetyassessment
Three Levels of Equivalence
Kesetaraan; Proksimat
Air Abu Protein Lemak0
10
20
30
40
50
60
%B
B
Tempe Kedele Import
Mean
Air Abu Protein Lemak0
10
20
30
40
50
60
%B
B
Tempe Kedele Lokal
Mean
Perbedaan signifikan : kadar air
Sumber: Syah (2005)
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%
C16:0(palmitic)
C18:0(stearic)
C18:1 cis(oleic)
C18:2(linoleic)
C18:3(linolenic)
0
10
20
30
40
509 field sites in 1992
%
Chemical CharacterisationFatty Acid Analysis of Seeds
*No significant differences from the control line were observed
Sumber: Hugget (2001)
Komposisi Asam Lemak (% AL/Total AL)
0
10
20
30
40
50
60
70
80
90
100
Palmitat Stearat Arakhidat Dodekanoat Oleat Linoleat Linolenat
Tempe Kedele Import Tempe Kedele Lokal
%AL/TotalAL
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H
leucine
tyrosine
phenylalanine
histidine
lysine
arginine
c
ysteine
methionine
tryptophan
0
1
2
3
%DryWeight
9 field sites in 1992
Control seeds
RRS seeds
Literature low
Literature high
*No significant differences from the control line were observed
Chemical CharacterisationAmino Acid Analysis of Seeds
Sumber: Hugget (2001)
Kesetaraan : Komposisi Asam Amino
0
1
2
3
4
5
6
As p Glu Se r His Gly Thr Arg Ala Tyr M et Val Phe Le u L ys
Tempe Kedele Import Tempe Kedele Lokal
%
BB
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Polar Chart Asam Amino (%AA/AA Total)
0
5
10
15
2025 Glu
Ser
His
Gly
Thr
Arg
Ala
Tyr
Met
Val
Phe
Leu
Lys
Tempe Kedele Import Tempe Kedele Lokal
Spektrum
Asam Amino
yang identik
Toxicant Analysis of Seeds fromRoundup Ready Soybeans
Trypsin inhibitor
TIU / mg DW
Lectin
H.U./mg protein extracted
0
20
40
60
80
100
%
Dryweight
9 field sites in 1992
Control seeds
RRS seeds
Literature low
Literature high
*No significant differences from the control line were observed Sumber: Hugget (2001)
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0
20000
40000
60000
80000
Kedele Tempe
Import Lokal
Kesetaraan : Tripsin Inhibitor
TIU/g
0,25
0,26
0,27
0,28
Kedele Tempe
Import Lokal
Kesetaraan : Asam Fitat
mg/g
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Kesetaraan : Hemaglutinin
Tempe Kedele Import (1681 HU/g)
Tempe Kedele Lokal (1648 HU/g)
No Changes in EndogenousAllergens
RR1: GTS line 61-67-1RR2: Parental line A5403RR3: GTS line 40-3-2C1: Cargill control 1C2: Cargill control 2C3: ADM control
106 kD
80 kD
49.5 kD
32.5 kD
27.5 kD
18.5 kD
Total ProteinSoyabean Sensitive
SerumNon-sensitive
Serum
RR-1
RR-2
RR-3
Control2
Control3
Control4
RR-1
RR-2
RR-3
Control2
Control3
Control4
RR-1
RR-2
RR-3
Control2
Control3
Control4
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Based on assessment of over 400 components in 2000 independent analyses -J. Nutr. (1996) 126:702-716
Confirmed with soybeans treated with Roundup herbicide - J. Agri. Food Chem.1999, 47:4469-4473
No changes in endogenous soya allergens - J. Allergy Clin. Immunol. 1996,96:1008-1010
Chemical Characterisation - Summary
Component Beans Meal Flour Isolate Conc. Oil
Proximate analysis = = = = =Amino acids =
Fatty acids = =
Trypsin inhibitors = = =
Lectins = =
Phytoestrogens = =
Urease = =
Stachyose, raffinose =
Phytate =
Known allergens = =
Component
Nutritional/Toxicant composition
Animal feed performance
Processing
Dietary use
Tolerance to glyphosate
One additional protein / gene
Unchanged
Changed
X
X
GM Soybeans vs Traditional Varieties
Sumber: Hugget (2001)
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Teknologi Pangan - IPB
Agenda
Pengantar
Substantial Equivalen
Beyond Substantial Equivalen
Pro Kontra
Pengaturan di Indonesia
Keamanan seluruh Komponen;
gen sisipan, pembawa dan inang
Tomato Fish
Tomato-fish
Replicated DNA
Denucleated fish egg
Fish cell
Fish DNATomato cell
Tomato DNA
Desired gene
Credit: Owen Koo
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Safety of Genetic Material
Inserted
Characterization of
Source
Size
Number
Location of insertion
Identification of sequences in the plantSafety of DNA itself is not in question
Safety of CTP-EPSPS
No toxicity associated with CTP-EPSPS gene
No toxicity associated with CTP-EPSPS protein- EPSPS family of proteins have a long history of safe
consumption
- No amino acid sequence homology to known toxinsor allergens
- No toxicity observed in animal studies
- Shows none of the properties of allergens
J. Nutrition, 1996, 126:728-740
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Males, pre-test Males, day 7, Females, pre-test Females, day 7
0
5
10
15
20
25
30
35
Meanbodyweight(g)
Vehicle control
363 mg/kg BSA control
49 mg/kg CP4 EPSPS
154 mg/kg CP4 EPSPS
572 mg/kg CP4 EPSPS
CP4 EPSPS - Toxicity Assessment
Digestion of CP4 EPSPS in vitro
CP4 EPSPS does not have the stabilitycharacteristics of known food allergens
Gastric fluid Intestinal fluid
0 15 30 60 120
SecondsC 0 10 32 100 270
MinutesC 1181
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Molecular weight (10-70 kD) + +High concentration in food + -Heat stable + -pH stable + -Protease stable + -Glycosylated + -Sequence homology + -
Comparison of CP4 EPSPS to knownallergenic proteins
no significant concern
Allergen CP4 EPSPS
Unintended Effects:
the unintended expression of someunknown or unexpected toxic orantinutrient factor, or the otherwiseunintended enhanced production ofknown toxic constituents. - (Royal Society,1998)
Evidence of such effects has not beenfound as result of bioengineering.
Effects are less likely in bioengineeredthan in conventional products.
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Food Allergens:
All food allergens are proteins: only a small
fraction of food proteins cause allergic
response.
Common foods with allergenic proteins:
peanuts, milk, seafoods
Potential allergenicity of genetic material
(proteins) introduced into a plant is an
important part of safety assessment.
Allergenicity Assessment
Follows a decision-tree process
Includes:
Source of the gene
Sequence homology of newly introducedprotein
Reactivity of newly introduced proteins withImmunoglobulin E (IgE) antibodies frompeople with known allergies to sourcematerial
Other properties, such as digestibility of theprotein
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Metode : Pengujian Alergenitas
Test Passive Cutaneous Anaphylaxis.- Sensitisasi mencit dengan pemberian ransum tempeselama 2 bulan.
- Mencit dianestasi, darahnya diambil, diperoleh serummencit anti-alergen
- Antiserum diinjeksikan secara intradermal dipunggungmencit
- Mencit diinjeksi lagi dengan sampel dan evans bluemelalui ekor
- Sampel menyebabkan alergi jika terlihat area biru padapunggung
Allergenitas ; PCA
Negatif
untukkedua
tempe
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Allergenitas ; Ig E
0Protein tempe kedelai lokal (3%)60Protein tempe kedelai lokal (1%)5
0.2Protein tempe kedelai import (3%)4
0Protein tempe kedelai import (1%)3
0.4Ovalbumin (1%)2
0Standar1
Hasil(IU/ml)
PerlakuanNo
Substantial
Equivalence Tox Testing
Pro Kontra (1)
Assumption based
risk assessment
SCIENCE/Evidence
based
risk assessment
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EVENT-Specific
Whole Food
Tox testing
Tox Testing
with Protein
(from E.coli)
Pro Kontra (2)
(Unproved) model
testing risk
assessmentTest as consumed
risk assessment
GM
Labelling
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Teknologi Pangan - IPB
Pengaturan di Indonesia
UU 7 Tahun 996 dan PP turunannya
Pasal dan Ayat Khusus
Labelling
Pengaturan
Komisi Keamanan Hayati dan Keamanan Pangan
Komisi Teknis Keamanan Hayati dan Keamanan
Pangan