Deteksi Keamanan Pangan

Transgenik
Dahrul Syah

Agenda

Pengantar
Substantial Equivalen
Beyond Substantial Equivalen
Pro Kontra
Pengaturan di Indonesia

Teknologi Pangan - IPB

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

4. Whole foods/crops
FLAVR SAVR,
Endless Summer
tomatoes
Virus resistant
squash
Insect resistant
potatoes, corn
Herbicide resistant
soybeans, canola

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
AGRONOMICS
PHENOTYPICS
GENOMICS
PROTEOMICS
METABOLOMICS
STATISTICS
Structure

Gene

Protein Metabolite

Function expression expression

expression

Data
integration

Unintended effects?

DIFFERENCES
COMPONENTS
PROTEINS
DNA/mRNAs
TISSUE
PLANT

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?

Equivalency testing
Selection of Traditional
Counterpart

History of safe use

Genetic characterisation
Agronomic characterisation

Comparison
with Traditional
Counterpart

Chemical characterisation
nutrients
toxicants
allergens
Dietary patterns

Kesetaraan Substansial
Asam Lemak

Allergens

Nutrients:

Carbohydrates
Proteins
Lipids
Vitamins
Minerals

Asam Amino

Tempe yang dibuat dari kedele import dan kedele lokal

Three Levels of Equivalence

1.Equivalent
Oil

No further testing

Oil

2.Equivalence except for

predictable, well-defined traits
Focused evaluation on limited
differences

3.Lack of equivalence, multiple

complex differences or no
traditional counterpart
Rigorous case-by-case safety
assessment

Kesetaraan; Proksimat
60

Mean

Mean

50

50

40

40

% BB

% BB

60

30

30

20

20

10

10

Air

Abu

Protein

Lemak

Air

Tempe Kedele Import

Abu

Protein

Lemak

Tempe Kedele Lokal

Perbedaan signifikan : kadar air

Sumber: Syah (2005)

Chemical Characterisation
Fatty Acid Analysis of Seeds
9 field sites in 1992

50

30

40

20
10
0

C16:0
(palmitic)

C18:0
(stearic)

C18:1 cis
(oleic)

C18:2
(linoleic)

C18:3
(linolenic)

*No significant differences from the control line were observed

Sumber: Hugget (2001)

% AL/Total AL

Komposisi Asam Lemak (% AL/Total AL)

100
90
80
70
60
50
40
30
20
10
0

Palm itat

Stearat

Arakhidat

Dodekanoat

Tempe Kedele Import

Oleat

Linoleat

Linolenat

Tempe Kedele Lokal

Chemical Characterisation
Amino Acid Analysis of Seeds
9 field sites in 1992

% Dry Weight

tryptophan

methionine

cysteine

arginine

lysine

histidine

tyrosine

leucine

phenylalanine

Control seeds

Literature low

RRS seeds

Literature high

*No significant differences from the control line were observed

Sumber: Hugget (2001)

Kesetaraan : Komposisi Asam Amino

6

% BB

5
4
3
2
1
0

Asp

Glu

Ser

His

Gly

Thr

Arg

Tempe Kedele Import

Ala

Tyr

Met

Val

Phe

Leu

Lys

Tempe Kedele Lokal

Polar Chart Asam Amino (%AA/AA Total)

Lys
Leu
Phe

25
20
15
10
5
0

Glu
Ser

Spektrum
Asam Amino
yang identik

His

Val

Gly

Met

Thr
Tyr

Ala
Tempe Kedele Import

Arg
Tempe Kedele Lokal

Toxicant Analysis of Seeds from

Roundup Ready Soybeans
100
9 field sites in 1992

% Dry weight

80
60
40
20
0

Trypsin inhibitor
TIU / mg DW

Lectin
H.U./mg protein extracted
Control seeds

Literature low

RRS seeds

Literature high

*No significant differences from the control line were observed Sumber: Hugget (2001)

10

Kesetaraan : Tripsin Inhibitor

60000
40000
20000
0
Kedele

Tempe
Import

Lokal

Kesetaraan : Asam Fitat

0,28
mg / g

TIU / g

80000

0,27
0,26
0,25
Kedele

Tempe
Import Lokal

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Kesetaraan : Hemaglutinin

Tempe Kedele Import (1681 HU/g)

Tempe Kedele Lokal (1648 HU/g)

No Changes in Endogenous
Allergens
Soyabean Sensitive
Serum

Total Protein

Non-sensitive
Serum
RR1: GTS line 61-67-1
RR2: Parental line A5403
RR3: GTS line 40-3-2
C1: Cargill control 1
C2: Cargill control 2
C3: ADM control

106 kD
80 kD
49.5 kD
32.5 kD
27.5 kD
18.5 kD
Control 4

Control 3

Control 2

RR-3

RR-1

RR-2

Control 4

Control 3

Control 2

RR-3

RR-2

RR-1

Control 3

Control 4

RR-3

Control 2

RR-1

RR-2

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Chemical Characterisation - Summary

Component
Proximate analysis
Amino acids
Fatty acids
Trypsin inhibitors
Lectins
Phytoestrogens
Urease
Stachyose, raffinose
Phytate
Known allergens

Beans

Meal

Flour

Isolate

Conc.

=
=
=
=
=
=
=
=
=
=

=
=

Oil

=
=
=
=

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

GM Soybeans vs Traditional Varieties

Component

Changed

Unchanged

Nutritional/Toxicant composition
Animal feed performance
Processing
Dietary use
Tolerance to glyphosate

One additional protein / gene

Sumber: Hugget (2001)

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Agenda

Pengantar
Substantial Equivalen
Beyond Substantial Equivalen
Pro Kontra
Pengaturan di Indonesia

Teknologi Pangan - IPB

Keamanan seluruh Komponen;

gen sisipan, pembawa dan inang
Tomato DNA

Fish DNA

Tomato cell

Fish cell
Desired gene

Replicated DNA

Fish

Tomato
Denucleated fish egg

Tomato-fish
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 plant
Safety 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 toxins
or allergens
- No toxicity observed in animal studies
- Shows none of the properties of allergens
J. Nutrition, 1996, 126:728-740

15

CP4 EPSPS - Toxicity Assessment

Mean body weight (g)

35
30
25
20
15
10
5
0
Males, pre-test

Males, day 7,

Females, pre-test

Vehicle control

49 mg/kg CP4 EPSPS

363 mg/kg BSA control

154 mg/kg CP4 EPSPS

Females, day 7

572 mg/kg CP4 EPSPS

Digestion of CP4 EPSPS in vitro

Gastric fluid

15

30 60 120

Seconds

Intestinal fluid

10

32 100 270 1181

Minutes

CP4 EPSPS does not have the stability

characteristics of known food allergens

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Comparison of CP4 EPSPS to known

allergenic proteins
Allergen

Molecular weight (10-70 kD)

High concentration in food
Heat stable
pH stable
Protease stable
Glycosylated
Sequence homology

+
+
+
+
+
+
+

CP4 EPSPS

+
-

no significant concern

Unintended Effects:
the unintended expression of some
unknown or unexpected toxic or
antinutrient factor, or the otherwise
unintended enhanced production of
known toxic constituents. - (Royal Society,
1998)

Evidence of such effects has not been

found as result of bioengineering.
Effects are less likely in bioengineered
than in conventional products.

17

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 introduced
protein
Reactivity of newly introduced proteins with
Immunoglobulin E (IgE) antibodies from
people with known allergies to source
material
Other properties, such as digestibility of the
protein

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Metode : Pengujian Alergenitas

Test Passive Cutaneous Anaphylaxis.

- Sensitisasi mencit dengan pemberian ransum tempe
selama 2 bulan.
- Mencit dianestasi, darahnya diambil, diperoleh serum
mencit anti-alergen
- Antiserum diinjeksikan secara intradermal dipunggung
mencit
- Mencit diinjeksi lagi dengan sampel dan evans blue
melalui ekor
- Sampel menyebabkan alergi jika terlihat area biru pada
punggung

Allergenitas ; PCA

Negatif
untuk
kedua
tempe

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Allergenitas ; Ig E
Hasil
(IU/ml)

No

Perlakuan

Standar

Ovalbumin (1%)

Protein tempe kedelai import (1%)

Protein tempe kedelai import (3%)

0.2

Protein tempe kedelai lokal (1%)

Protein tempe kedelai lokal (3%)

0
0.4

Pro Kontra (1)

Assumption based
risk assessment

Substantial
Equivalence

SCIENCE/Evidence
based
risk assessment

Tox Testing

20

Pro Kontra (2)

(Unproved) model
testing risk
assessment

Tox Testing
with Protein
(from E.coli)

Test as consumed
risk assessment
EVENT-Specific

Whole Food
Tox testing

GM
Labelling

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Pengaturan di Indonesia
UU 7 Tahun 1996 dan PP turunannya
Pasal dan Ayat Khusus
Labelling
Pengaturan
Komisi Keamanan Hayati dan Keamanan Pangan
Komisi Teknis Keamanan Hayati dan Keamanan
Pangan
Teknologi Pangan - IPB

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