BACTERIAL CELLULAR SUBSTANCE 85 



Glutamic acid was separated as the hydrochloride. 

 Chlorides of ammonium and sodium were present in large 

 amount, but were easily separated on account of their 

 greater solubility in water. We obtained 1 gram of glutamic 

 acid, equivalent to 0.2 per cent, of the cellular substance 

 or 0.23 per cent, of the moisture-free substance. 



The filtrate from the glutamic acid on being esterified, the 

 esters extracted, dried, and distilled, yielded the following: 



Temperature Temperature Weight of 



The pasty mass left after extraction of the esters was 

 acidified with hydrochloric acid, evaporated, the salt 

 filtered out from time to time, and when brought to a thick 

 syrup it was diluted with an equal volume of absolute 

 alcohol and the esterification repeated. However, the yield 

 from this esterification was exceedingly small, not more 

 than a few drops for each fraction. 



From the residue left after distillation, we obtained 2 

 grams of leucinimide, equivalent to 0.4 per cent, of the 

 cellular protein or 0.45 per cent, of the moisture-free sub- 

 stance. 



Fraction I. This was saponified by being boiled with 

 five times its weight of water for five hours under a reflux 

 condenser. It was evaporated to dryness and the white 

 crystalline mass was dissolved in 25 c.c. of hot water, 

 treated with an equal volume of hot absolute alcohol and 

 left in the ice-box for two days when a white crystalline 

 mass separated. This was purified and yielded 7 grams of 

 alanin, equivalent to 1.4 per cent, of the cellular substance 

 or 1.57 per cent, of the moisture-free substance. 



Fraction II. From this there was obtained by the method 

 already described a copper-leucin-valin compound repre- 



