WORLD HEALTH ORGANIZATION
WHO Food Additives Series 1972, No. 1
TOXICOLOGICAL EVALUATION OF SOME
ENZYMES, MODIFIED STARCHES AND
CERTAIN OTHER SUBSTANCES
The evaluations contained in this publication were prepared by the
Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
16-24 June 19711
World Health Organization
1 Fifteenth Report of the Joint FAO/WHO Expert Committee on Food
Additives, Wld Hlth Org. techn. Rep. Ser., 1972, No. 488; FAO
Nutrition Meetings Report Series, 1972, No. 50.
MIXED MICROBIAL CARBOHYDRASE AND PROTEASE
This enzyme preparation is prepared from some varieties of the species
Data are available for some commercial preparations.
Animal Route LD50 Reference
mouse oral >3 200 Willard et al., 1971
rat oral >13 800 Osaka City Inst. Hyg. 1959
>10 000 Hartnagel and Phillips, 1969
>30 000 (amylase Kyoto College Pharm. 1957
>15 000 (amylase) " " " 1963
>15 000 (protease) " " " 1963a
Three groups of 10 male and 10 female rats received daily for 90 days
either 0 or 1 per cent. enzyme or 1 per cent. heat-inactivated enzyme
in their diet. No differences from controls were seen as regards
appearance, behaviour, body-weight, food intake, organ weights,
haematology, urinalysis, gross and histopathology (Gesler & Jennings,
In another study 4 groups of 10 male rats received daily for 5 days by
gavage enzyme (Bax 1092) at 0, 0.25, 1.0 and 4.0 g/kg body-weight and
observed for another 7 days. Growth rate, appearance and behaviour
were similar in all groups. Haematology, organ weights and gross
pathology showed no abnormalities between the groups (Garvin et al.,
B. subtilis occurs ubiquitously and is a common contaminant of food.
The available information appears to indicate that it is not
pathogenic in man. An adequate 90-day study in rats on one strain is
available as well as another study extending over 12 days with a
maximum level of 4 g/kg. It was considered that enzyme preparations
from this organism carry no toxicological hazard.
Garvin, P. J., Merubia, J., Salazar, M., Seagraves, P., Willard, R. &
Lehman, M. (1962) Unpublished report supplied by Baxter Laboratories
Gesler, R. M. & Jennings, R. B. (1958) Unpublished report submitted by
Baxter Laboratories Inc.
Hartnagel, R. E. and Phillips, B. M. (1969) Unpublished report
submitted by Miles Chem. Co.
Kyoto College of Pharmacy (1957) Report on Biotamylase submitted to
Kyoto College of Pharmacy (1963) Report on Biotamylase A1000 submitted
Kyoto College of Pharmacy (1963a) Report on Biotamylase P1500
submitted to WHO.
Osaka City Institute of Hygiene (1959) Report on bioprase SP-10
submitted to WHO.
Osaka City Institute of Hygiene (1966) Report on bioprase PN-conc
submitted to WHO
Willard, R. L., Oskorep, D. & Garvin, P. J. (1971) Unpublished report
submitted by Travenol Laboratories Inc.
* Except for good manufacturing practice.