Guidelines for Safety Assessment of Foods and Food Additives Produced by the Recombinant DNA Techniques (draft revision)

Eubios Journal of Asian and International Bioethics 6 (1996), 21.


For questions or comments regarding these draft guidelines contact, K. Tanaka, MHW, Japan (Email: PXE00135@niftyserve.or.jp)
Chapter 1. General Provisions
1. Objectives. The objectives of the Guidelines are to establish basic requirements for safety assessment of foods and food additives produced by recombinant DNA techniques and thereby to ensure the safety of these foods and food additives.
2. Definitions of terminology. The terms used in the Guidelines are defined as follows:
1) Recombinant DNA techniques. A technique by which DNA is enzymatically cleaved, recombined and transferred to living cells (host) for proliferation.
2) Host. Living cells to which DNA is transferred through recombinant DNA techniques.
3) Vector. Carrier DNA which transfers the target genes into hosts for proliferation and expression
4) Inserted DNA (or gene). Foreign DNA (or gene) inserted into vectors.
5) Recombinant. Viable cells containing recombinant DNA
6) Gene products. Any substance derived from the inserted gene.
7) Product. Substances produced by recombinant DNA techniques.
3. Scope of Application. The Guidelines are applicable to the products which are regarded as equivalent to the existing products used as foods and food additives. When the product containing the recombinant is to be consumed, the Guidelines are applicable only if the recombinant is speratophytes. Whether or not the products are regarded as equivalent to the existing products is assessed based upon the following information (1) to (4): (1) Information on the genetic material (2) Information on broad human consumption history (3) Information on components of foods (4) Information on difference in usage between the conventional varieties and the new varieties

Chapter 2. Safety assessment of manufacturing process
When the recombinant is not to be consumed, the safety of manufacturing process should be assessed in order to ensure the safety of the recombinant.Manufacturing method of the recombinant (including preparation method and manufacturing facilities), the raw materials, and the equipments used will be evaluated.
1. Manufacturing methods of the recombinant (Manufacturing methods and Facilities). It is the fundamental principle to use a non-pathogenic recombinant in accordance with GILSP (Good Industrial Large Scale Practice) or Category I. The safety is assessed based upon the safety assessment items for the recombinant specified in Attachment 1.
2. Raw materials other than recombinants and equipments used for manufacturing. The safety is assessed based upon the following information (1) and (2): (1) Information on history of usage of the raw materials and the equipment used for manufacturing the foods and food additives. (2) Information on safety of the raw materials and the equipment used for manufacturing the foods and food additives or examination results specified in Attachment 3.
3. Purification of the products. Safety is assessed based upon the product purification method and its effectiveness.

Chapter 3. Safety assessment of the product
Safety assessment is made in terms of all factors resulted in alteration by using recombinant DNA techniques.

1. Safety assessment in cases where the recombinant is not to be consumed
1) Safety assessment is made based upon the following information (1) to (4):(1) Information demonstrating the lack of contamination by the recombinants(2) Information concerning the safety of impurities derived from the manufacturing process(3) Information on Chapter 2. 3 concerning the product purification method(4) Information on changes of constituents which posses harmful effects if their levels are changed.
2) If safety of the product is not well established based upon the items listed in 1), safety should be assessed by examination results specified in Attachment 3.

2. Safety assessment in cases where the recombinant is to be consumed
1) Safety assessment is made based upon the items specified in Attachment 2 (including Appendixes 1 and 2).2) If safety of the product is not well established based upon the items listed in 1), safety should be assessed by examination results specified in Attachment 3. A nutritional test may be required if needed.

Chapter 4. Confirmation by the Minister of Health and Welfare
Those who wish to manufacture or import foods and food additives produced by recombinant DNA techniques may request the Minister of Health and Welfare confirmation regarding compliance of the products to the Guidelines.


(Attachment 1)
Necessary information for safety assessment when the recombinant is not to be consumed
(1) Purposes and usage of recombinants
(2) Host
a. Taxonomy (scientific name and strain name)
b. Production of pathogenic or harmful, physiologically active substances
c. Parasiticity and fixing conditions
d. Foreign factors (virus, etc.) (a host should not be contaminated with pathogenic exogenous factors.)
e. Survival and proliferation abilities under experimental conditions simulating ordinary or natural environments
f. Sexual or asexual reproduction periods and crossing
g. Historical background regarding the application of foods or food additives
h. Restrictive conditions on survival and propagation abilities
i. Pathogenicity of host-related strains and production of harmful physiological active substance
(3) Vector
a. Name
b. Its origin
c. Its properties
c-1. DNA molecular weight
c-2. Cleavage map, using restriction enzyme
c-3. Presence of nucleic acid sequence to code for a known toxin
d. Drug resistance
e. Transmissionality
f. Host dependency
g. Method for developing the expression vector
h. Introduction method of the expression vector into the host and its location in the host
(4) Inserted gene (DNA)
1) Donor
a. Name and classification
2) Inserted gene
a. Structure
a-1. Presence of nucleic acid sequence to code for a known toxin
b. Properties
b-1. Function of inserted genes
b-2. Cleavage map by restriction enzyme
b-3. DNA molecular weight
(5) Recombinants
a. New properties acquired by recombinant DNA techniques (non-pathogenic)
b. Survival and proliferation abilities in the natural environment
c. Restriction conditions for survival and proliferation abilities of host*1
d. Inactivation methods of the recombinant
e. Differences from the host*2
Note
1. For those recombinats listed in the Table 2 or 3 of "Guidelines for Recombinant DNA Experiments", some of information above which is already known may be exempted.

2. Criteria of properties of recombinants which can be used for manufacturing in accordance with GILSP (Good Industrial Large-Scale Practice) or Category 1 are based upon the OECD report, "Recommendations for safety consideration for industrial, agricultural, and environmental applications of organisms derived from recombinant DNA techniques" (1986).

*1In industrial utilization, recombinants should be as safe as the host, shoul d exhibit limited propagation in external environments and should not give any harmful effects to environments.
*2It is required to attach information on absence of pathogenicity and of production of harmful physiologically active substances.


(Attachment 2)
Necessary information for safety assessments when the recombinant is to be consumed

(1) Purposes and usage of recombinants
(2) Host
a. Classification (Scientific name, species and strain)
b. Genetic origin
c. Production of harmful physiologically active substances
d. Allergenicity
e. Parasiticity and fixing conditions
f. Foreign factors (virus etc.) (a host should not be contaminated with pathogenic exogenous factors.)
g. Survival and propagation under experimental conditions simulating ordinary or natural environments
h. Sexual reproduction periods and crossing
i. History of utilization as food
j. Safe consumption
k. Restrictive conditions on survival and propagation abilities
l. Pathogenicity of and production of harmful physiologically active substances in ancestral or related species to the hosts

(3) Vector
a. Name
b. Its origin
c. Its properties
c-1. DNA molecular weight
c-2. Cleavage map, using restriction enzyme
c-3. Presence of nucleic acid sequence to code for a known toxin
d. Drug resistance
e. Transmissionality
f. Host dependency
g. Preparation method of expression vector
h. Introduction method of the expression vector into the host and its location in the host

(4) Inserted gene (DNA)
1) Donor
a. Name, origin, and taxonomy
b. Safe consumption
2) Inserted gene (DNA)
a. Information concerning structure
a-1. Promoter
a-2. Terminator
a-3. Presence of nucleic acid sequence to code for a known toxin
b. Properties
b-1. Function of the inserted genes
b-2. Cleavage map by restriction enzyme
b-3. DNA molecular weight
c. Purity
d. Stability*1
e. The number of copies of the introduced genes*1
f. The sire, timing, and amounts of expression*1
g. Safety on antibiotic-resistant markers*1 *2
h. Presence or absence of exogenous open reading frames and the possibility of their transcription and expression*1
(5) Recombinants
a. New properties acquired by recombinant DNA techniques
b. Allergenicity of recombinant products*3
c. Toxicity of recombinant products (other than allergenicity)
d. Effect of recombinant products on metabolic pathways*4
e. Difference from the host*5
f. Survival and propagation in the natural environments
g. Restriction condition for survival and propagation of the recombinants
h. Inactivation method of the recombinant
i. Approval and usage as food in other countries
j. Methods of preparation, breeding, and cultivation
k. Method of seed production and management

*1 Any changes which occur within the recombinant should be considered.
*2 Information should be prepared according to Appendix 1.
*3 Information should be prepared according to Appendix 2.
*4 Information concerning possibility of reacting with endogenous substances in the conventional species should be prepared.
*5 Information concerning nutrients, anti-nutrients and constituents which may cause harmful effect when the level is changed.


(Attachment 2, Appendix 1)
Information necessary for safety assessment of antibiotic-resistant marker genes

Safety of antibiotic-resistant marker genes is assessed according to the following information (1) and (2):
(1) Information on characteristics of genes and their products
- Structure and function
- Mechanism of the manifestation of the resistance, the method of use, and related metablities
- Method for identification and quantification
- Changes accompanying cooking or processing (stability against heat and physical pressures)
- Changes in gastrointestinal tract environments (stability against acid and digestive enzymes)

(2) Information concerning consumption of genes and their products
- Expected amounts of consumption
- Present usage of antibiotics associated with resistance
- Comparison with the amount and characteristics of antibiotics-reistant bacteria that are normally present
- Estimated amount of antibiotics inactivated after oral administration and the possibility that inactivation may cause problems


(Attachment 2, Appendix 2)
Necessary information for safety assessment of allergenicity
Safety concerning allergenicity is assessed according to the following information (1) to (6):
(1) Consumption history of the donor
(2) Whether the gene product is known as an allergen
(3) Sensitivity of the gene product to physicochemical treatment*1
(4) Whether it causes significant change in the amount of the gene product ingested
(5) Structural homology of gene products with known food allergens
(6) Whether the gene product occupies a considered part in the total protein ingestion per day.

1. Some information may be exempted if there are valid rationales.
2. If safety is not confirmed on the basis of (1) to (6), submission of the following data and consultation with the Ministry of Health and Welfare are required.
- Information concerning whether the gene product binds to IgE antibody from patients sensitive to homologic allergens*2
- Information concerning whether the gene product binds to IgE antibodies
from patients sensitive to major antigens*2 *3
*1 Sensitivity to treatment with artificial gastric juice, artificial intesti nal juice, and heat treatment should be examined by protein electrophoresis and the Western blot method.
*2 Capacity binding to IgE antibody from allergic patients should be examined by the Western blot method and ELISA method.
*3 Tests should be conducted using sera from patients allergic to egg, milk, soybeans, rice, wheat and buckwheat.
(Attachment 3)
(1) Acute toxicity study
(2) Subacute toxicity study
(3) Chronic toxicity study
(4) Tests for influences on reproduction
(5) Mutagenicity study
(6) Carcinogenicity study
(7) Other necessary tests (intestinal tract toxicity test, etc.)

1.Test data should be obtained by studies conducted at approved GLP (Good Laboratory Practice) facilities in accordance to the GLP and the major part of the test results should have been published in scientifically referred journals.

2.Some information may be exempted if there are valid rationales.
Attachment 1 Necessary information for safety assessment when the recombinant is not to be consumed
Attachment 2 Necessary information for safety assessments when the recombinant is to be consumed
Appendix 1 Necessary information for safety assessment of antibiotic- resistant markers
Appendix 2 Necessary information for safety assessment of allergenicity
Attachment 3



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