598 Dr. E. F, Armstrong. [July 29, 
glucosazone was observed after 20 minutes’ heating; in the control experi- 
ment no separation took place but a soluble osazone was formed. 
Proof of the Presence of Maltose-—In order to destroy the glucose, the liquid 
was fermented with S. Marxianus, a yeast which does not contain maltase 
and therefore is without action on maltose. After removal of the fermentable 
matter, the solution, 200 e.c. in volume, containing the product from 
50 grammes of maltose, had the rotatory power ep = +7°35, which was 
about double that observed in a similar solution containing isomaltose alone. 
The osazone formed on heating with phenylhydrazine was entirely soluble in 
boiling water, but distinctly less soluble than the isomaltosazone; it began to 
separate from the liquid while this was still hot and the osazone could be 
fractionated into more and less soluble portions. 
Separate portions of the solution were digested with emulsin and maltase ; 
in both cases glucoses were formed. When subjected to the joint action of 
emulsin and a yeast containing maltase (S. intermedians), the solid matter in 
solution disappeared almost entirely in a single fermentation. 
The isomaltose is apparently present in larger quantity than maltose: it is 
proposed to determine their relative proportions by removing the isomaltose 
from the mixture by the joint action of emulsin and S. Marxianus. 
The Production of Isomaltose by means of Maltase. 
The maltase extract used was always prepared by grinding 5 grammes of 
air-dried top yeast (from a London brewery) with 100 cm. of water and then 
digesting the mixture at 25° during two to three hours. Fifty grammes of 
glucose were dissolved in about 75 c.c. of the filtered extract, and some 
toluene was added to maintain the liquid sterile; the solution was kept 
in a stoppered bottle at 25° during two to three months or even longer; it 
darkened somewhat but remained perfectly clear and sweet. To remove 
glucose and maltose, the solution diluted with an equal volume of water was 
boiled with charcoal and filtered ; yeast water was then added and after the 
liquid had been sterilised it was fermented with S. intermedians. 
To insure complete removal of the glucose, it was necessary to repeat the 
fermentation at least twice. Ultimately, a clear solution was obtained which 
not only had a strong reducing action on Fehling’s solution but also a high 
positive rotatory power. When treated with phenylhydrazine in the manner 
described, it gave an osazone in every way identical with isomaltosazone ; in 
fact, when the experiments were carried out side by side with the products 
obtained by means of acid and by means of the enzyme, no difference could 
be detected. The product also behaved in the manner to be expected towards 
emulsin, being converted into glucose. 
