30 
solved in 20 c.c. water and filtered. Five c.c. of the filtrate 
were placed in each of three 100 c.c. flasks, along with ‘2 gm. of the. 
protein and 25c.c. of water. The flasks were treated as described 
above,and after precipitation with tannic acid, made up to 100 c.c. 
with water, and the contents were filtered. 
The nitrogen was in each case determined in 30 c.c. of the 
filtrate, with the following results :— 
POLYPORUS SQUAMOSUS. 
(a) Flask with proteolytic activity gave . 3°5 CC. ee 
(d) a) ” a) 4°0 Cc Cc 3? >? 
(c) x NO 7 ‘3 5 SO-6.0h ay halk 
POLYPORUS SULPHUREUS. 
(a2) Flask with proteolytic activity gave Paeg Oe. Sir, 
(0) a? 23 a7 - 370 C.C. »”»> a”? 
(c) eo OS rs Fh go ia / 
MERULIUS LACRYMANS. 
(a) Flask with proteolytic activity gave 7 a7 et. = NH, 
(d) ” a3 ” . a5 c.C. > x” 
(c) peeps &, 55 : a me eae ng 
These results show clearly that active proteolyses has been 
going on in all three cases. 
The accuracy was not satisfactory, especially in the case of 
P. sulphureus, but this was found to be due, not to the 
method, but to minor errors in the manipulations; in all the 
subsequent determinations the results of the duplicates have 
come out practically the same. 
In order to test the effect of a slight trace of acid or alkali 
on the activity the following experiment was carried out with 
the enzymatic preparation obtained from P. squamosus. 
‘5 gm. of the preparation was dissolved in 30 c.c. water, filtered, 
and 3 c.c. of filtrate put into each of nine test tubes along with 
‘2 gm. of protein. To the first three tubes hydrochloric acid 
was added, so that in 6 c.c. of the resulting solution the acidity 
would be equal to ‘xr per cent. To the second three tubes sodium 
carbonate was added, so that in 6 c.c. of resulting solution the 
degree of alkalinity would be equivalent to that of acidity in 
the first three. To the last three tubes 3 c.c. of water were 
added. 
