104 
Vines . — The Proteases of Plants (V). 
Ungerminated seed of Cannabis sativa. 
EXPERIMENT i. In this case a mixture of crushed seed with water was 
used ; all subsequent experiments were made with extracts of the seed. 
15 grms. of crushed seed were placed in each of two bottles with 100 cc. 
distilled water ; a sample of the liquid gave no tryptophane-reaction : to No. 1 nothing 
was added ; to No. 2, 0*2 grm. of well-washed fibrin. After 20 hours in the incubator 
No. 1 gave distinct tryptophane-reaction ; No. 2 gave a marked reaction, and the 
fibrin had entirely disappeared : 0-5 grm. fibrin was then added to No. 2. After 
48 hours’ digestion both gave strong tryptophane-reaction ; the 0*5 grm. fibrin added 
to No. 2 disappeared in 48 hours. Subsequently 0-3 grm. fibrin was added to No. 1 
and was digested within 24 hours. 
These results prove that autolysis proceeded actively during the experi- 
ment, and that the mixture was able to digest fibrin : it may be inferred 
that the resting seed contains both peptonizing and peptolyzing enzymes. 
It should be explained that, in such experiments as these, it is difficult to 
demonstrate the actual formation of peptones, for the liquid gives the 
biuret-reaction to begin with, owing to the presence of albumoses extracted 
from the seed. 
Some of these seeds were germinated with a view to comparative experi- 
ments with the ungerminated seeds and the seedlings ; but they were not 
pursued when the seeds were found to be so active proteolytically. The 
only observation made was that a watery extract of the seedlings, a week 
old, gave a strong tryptophane-reaction. 
Experiment 2. 10 grms. of seed were crushed with 100 cc. distilled water, and 
the same quantity with 100 cc. 2% NaCl-solution : the mixtures were left to filter in 
the cold all night. The filtered extracts were found to be acid, and to give a pre- 
cipitate of proteid on boiling, rather more in the NaCl-extract ; the NaCl-extract also 
gave a faint tryptophane-reaction. 
About 30 cc. of each extract were put separately into each of two bottles ; to one of 
the H 2 0 -extract bottles (1) and to one of the NaCl-bottles (3) nothing was added but 
some HCN (about o*i%) as an antiseptic ; to the other H ? 0 -bottle (2) and to the 
other NaCl-bottle (4) 0-2 grm. fibrin was added. After 24 hours’ digestion the fibrin 
in the NaCl-bottle had disappeared ; and after 48 hours that in the H 2 0 -bottle had 
disappeared. The precipitate on boiling the liquids was observed to diminish 
gradually. At the close of the experiment (96 hours) the precipitate was very slight in 
the two H 2 0 -bottles, rather more in the two NaCl-bottles ; the tryptophane-reactions 
were distinct in all the bottles, except No. 4, in which it was strong. 
The diminution of the precipitate on boiling affords evidence of digestive 
action on the coagulable proteids dissolved in the extract, that is of autolysis. 
The results confirm those of Experiment 1, and further show that the 
proteases of the seed can be dissolved out by both water and dilute NaCl- 
solution, apparently rather more by the latter than by the former. 
