ENZYMES DERIVED FROM ASPERGILLUS ORYZAE
EXPLANATION
Enzymes from this source were considered at the fifteenth meeting
of the Committee (Annex 1, reference 26), at which time a decision on
the ADI was postponed because of concern that one of the known
metabolites of A. oryzae is ß-nitropropionic acid, which was
suspected of carcinogenic potential. Later, at the eighteenth meeting
of the Committee, a lipase derived from this organism was considered
(Annex 1, reference 35). It was determined at that time that there was
no information to substantiate the concern for the potential
carcinogenicity of ß-nitropropionic acid, and that analyses of foods
have shown that the metabolite is present in very few foods and then
only in minute amounts. The present Committee was also informed that
A. oryzae varieties are used in certain parts of the world in the
preparation of foods.
alpha-AMYLASE (E.C. 3.2.1)
BIOLOGICAL DATA
Biochemical aspects
No information available.
Toxicological studies
Acute toxicity
Animal Route LD50 Reference
Mouse (Novo Strain) Oral > 20 g/kg b.w. Novo, 1971a
Short-term studies
Rats
Three groups, each containing 5 male and 5 female SPF Wistar
rats, were maintained for 3 weeks on diets containing 0, 0,5, or 5% of
the enzyme preparation. Only minor differences were observed among the
groups in body-weight change and food intake. At termination of the
study, haematologic measurements, organ-weights analyses, and gross
post mortem examinations showed no compound-related effects
(Novo, 1971b).
In another study, two groups, each containing 10 male and 10
female ARS Sprague-Dawley rats, were fed diets containing 5 or 10% of
the test enzyme (equivalent to 3.5 or 7 g enzyme/kg/b.w./day) for
90-94 days. A control group of 20 male and 20 female rats was
maintained on the diet alone. No signs of toxicity were observed
during the test period. Body-weight gain and food consumption were
similar among animals in the test and control groups. Differential
blood counts were within the normal range at weeks 4 and 8 in all
groups. At the end of the study, haematologic parameters, organ-weight
analyses, and gross and microscopic pathology showed no
compound-related effects (Garvin et al., 1972a).
A similar study was performed with carbohydrases from A. oryzae
(alpha-amylase and amyloglucosidase), prepared under different culture
conditions. No compound-related effects were reported
(Gavin et al., 1972b).
Long-term studies
No information available.
Observations in man
No information available.
COMMENTS
Short-term studies on alpha-amylase from A. oryzae did not
reveal any adverse effects. Based upon its lack of toxicity and the
fact that A. oryzae varieties are used in the preparation of foods,
this enzyme was considered to be acceptable for use in food.
EVALUATION
Level causing no toxicological effects
Rat: 10% in the diet, equivalent to 7 g/kg b.w./day.
Estimate of acceptable daily intake
Acceptable for use in food when used according to good
manufacturing procedures.
REFERENCES
Garvin, P.J., Ganote, C.E., Merubia, J., Delahany, E., Bowers, S.,
Varnado, A., Jordan, L., Hatley, G., DeSmet, C., & Porth, J. (1972a).
Unpublished report from Travenol Laboratories, Inc., Morton Grove,
IL, USA. Submitted to WHO by Gist-brocades NV, Delft, Holland.
Garvin, P.J., Ganote, C.E., Merubia, J., Delahany, E., Varnado, A.,
Jordan, L., Hatley, G., DeSmet, C., & Porth, J. (1972b). Carbohydrase
from A. oryzae. Unpublished report from Travenol Laboratories, Inc.,
Morton Grove, IL, USA. Submitted to WHO by Gist-brocades NV,
Delft, Holland.
Novo (1971a). Acute toxicity of fungamyl to mice. Unpublished report
from Novo Industri A/S, Bagsvaerd, Denmark. Submitted to WHO by Novo
Industri A/S, Bagsvaerd, Denmark.
Novo (1971b). Three week oral toxicity study of fungamyl in rats.
Unpublished report BSi/BS from Novo Industri A/S, Bagsvaerd, Denmark.
Submitted to WHO by Novo Industri A/S, Bagsvaerd, Denmark.
PROTEASES (B.C. 3.4.21.14; 3.4.23.6)
BIOLOGICAL DATA
Biochemical aspects
No information available.
Toxicological studies
Acute toxicity
No information available.
Short-term study
Rats
Two groups of 10 male and 10 female ARS Sprague-Dawley rats were
fed diets containing 5 or 10% of the test enzyme preparation
(equivalent to 3.5 or 7 g enzyme preparation/kg b.w./day) for 90 to 94
days. A control group of 20 male and 20 female rats were maintained on
the diet alone. No signs of toxicity were observed during the test
period. Body-weight gain and food consumption were similar in animals
in the test and control groups. Differential blood counts were within
the normal range at weeks 4 and 8 in all groups. At the end of the
study serum clinical chemistry parameters, organ weight analyses, and
gross and microscopic pathology showed no compound-related effects
(Garvin et al, 1972).
Long-term studies
No information available.
Observations in man
No information available.
COMMENTS
A short-term study in rats on a protease preparation from
A. oryzae did not reveal any adverse effects. Based on its lack of
toxicity and the fact that A. oryzae varieties are used in the
preparation of foods, this enzyme was considered to be acceptable for
use in food.
EVALUATION
Level causing no toxicological effect
Rat: 10% in the diet, equivalent to 7 g/kg b.w./day.
Estimate of acceptable daily intake
Acceptable for use in food when used according to good
manufacturing procedures.
REFERENCES
Garvin, P.J., Ganote, C.E., Merubia, J., Delahany, E., Bowers, S.,
Varuado, A., Jordan, L., Harley, G., DeSmet, C., & Porth, J. (1972).
Protease from Aspergillus oryzae. Unpublished report from Travenol
Laboratories, Inc., Morton Grove, IL, USA. Submitted to WHO by
Gist-brocades NV, Delft, Holland.