X. A, 2 



Brill: Enzymes of Cacao 



125 



EXPERIMENTAL 



A 10 per cent aqueous solution of the pulp and seed of cacao 

 was made by grinding them in a mortar with clean sand under 

 water, allowing them to stand two hours, and afterwards filtering 

 through cheese cloth. The seed was freed from the pulp and 

 surrounding slime by very careful washing in running water 

 and later removal of the inclosing membrane. Three series of 

 experiments were run: namely, with the surrounding pulp, the 

 fresh clean seed, and the clean fermented seed. The last was 

 prepared by placing the seed in the incubator and allowing the 

 process of fermentation to proceed for four days. It was then 

 removed, very carefully separated from the outer slime and 

 surrounding membrane, and treated as the other two. In all 

 cases toluene was used as the antiseptic. 



Tests were then carried out with these solutions for the de- 

 tection of enzymes. 



Table I. — Tests for lipase. 



SERIES 1. 



15 cc. water, 10 cc. pulp exti-aet, 48 hours at 38° C 



15 cc. water, 10 cc. pulp extract, 1 cc. olive oil, 48 hours at 38° C 



15 cc. water, 10 cc. pulp extract heated, 1 cc. olive oil, 48 hours at 38° C. 

 25 cc. water, 1 cc. olive oil, 48 hours at 38° 



Cc. N/10 

 NaOH. 



0.40 

 0.45 

 0.42 

 0.45 



SERIES 2. 



16 cc. water, 10 cc. seed extract, 48 hours at 38° C 



15 cc. water, 10 cc. seed extract, 1 cc. olive oil, 48 hours at 38° C 



16 cc. water, 10 cc. seed extract heated, 1 cc. olive oil, 48 hours at 38° C . 



0.40 

 0.40 

 0.45 



SERIES 3. 



10 cc. water, 10 cc. fermented seed extract, 48 hours at 38° C 



0.72 

 0.72 

 0.78 



15 cc. water, 10 cc. fermented seed extract, 1 cc. olive oil, 48 hours at 38° C -- 



15 cc. water, 10 cc. fermented seed extract heated, 1 cc. olive oil, 48 hours at 38° C 



No lipase was present that reacted on olive oil, as the control 

 r-equired the same amount of N/10 NaOH for neutralization as 

 the active sample. Duplicates of the above were carried out 

 substituting ethyl butyrate for olive oil with the like negative 

 results. 



