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

    Geneva

    1972





                   

    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

    Biological data

    Source

    This enzyme preparation is prepared from some varieties of the species
    Bacillus subtilis.

    Acute toxicity

    Data are available for some commercial preparations.

                                                                       

    Animal  Route          LD50            Reference
                     (mg/kg body-weight)
                                                                       

    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
                        & protease) 
                     >15 000 (amylase)       "      "      "    1963
                     >15 000 (protease)      "      "      "    1963a
                                                                       

    Short-term studies

    Rat

    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,
    1958).

    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.,
    1962).

    Long-term studies

    None available.

    Comments

    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.

    EVALUATION

    Not limited.*

    REFERENCES

    Garvin, P. J., Merubia, J., Salazar, M., Seagraves, P., Willard, R. &
    Lehman, M. (1962) Unpublished report supplied by Baxter Laboratories
    Inc.

    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
    WHO.

    Kyoto College of Pharmacy (1963) Report on Biotamylase A1000 submitted
    to WHO.

    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.
    

    See Also:
       Toxicological Abbreviations