﻿1 90^] EXPERIMENTAL STUDIES ON INULASE 25 



He says nothing concerning the inulase of Penicillium bej'ond 

 the bare fact of its existence. 



Chevastelon-^ was able to cause hydrolysis of the *' inulin" from 

 the monocotyledons by the agency of the inulase of Aspergillus, 



It seemed of interest to investigate more fully the action of 

 the inulase of Aspergillus and Penicillium, and to discover, if 

 possible, whether or not it was identical with the inulase of the 

 artichoke. 



CULTURE METHODS. 



Pure cultures of Aspergilhts iiiger and Penicillittm glatiaimv^^x^ 

 obtained, and stock cultures kept on the following medium : 



Medium A. — Agar-agar, lo^r™; NaCl, 2.5^; peptone (Grubler's), 2.5^^; 



beef extract, i^™; inulin (air dry), 5^"^; water, 500^*=. 



The inulin used in making all culture media was prepared 

 ixom Dahlia variabilis by alcoholic precipitation. 



The first experiments were carried out upon Penicillium. A 

 culture fluid having the following composition was prepared: 



MEDiUMB.—InuHn, 1. 5^"*; peptone (Griibler's), 0.5^"^ ; NaCl, 0.5s™ ; beef 

 extract, trace; water to make 150^^ 



Test-tubes containing this liquid were sterilized and inoculated 

 with the spores of Penicillium. The fungus grew well in this 

 medium. Each day the culture fluid of one tube was used to 

 test for the presence of sugar (levulose) in solution. During 

 the first three days no sugar reaction was evident, but after that 

 time a reduction could be obtained by boiling the culture fluid 

 with Fehling's solution. * The amount of sugar was small at 

 first, but steadily increased, until, after a week's time, the reac- 

 tion was very marked. 



Five days after inoculation the fluid from several cultures 

 was filtered and heated in a water bath with phenyl-hydrazine 

 ^hydrochloride and sodium acetate. The osazone formed sepa- 

 rated while the solution was in the boilincf water bath and had 



th 



e 



characteristic crystalline form of glucosazone. 

 In order to establish the fact that the inulin of the medium 

 was the source of the sugar formed, the following culture fluid 

 was prepared which contained no organic compound except inulin: 



*Sur rinuline d'ail, de la jacinthe, d'asphod^Ie et de la tubereuse. Jour. Pharm. 

 iV. a: I, 1895. 



