DOMOLD SPORES CONTAIN ENZYMS ? 



By Nicholas Kopeloff and Lillian Kopeloff ' 



Bacteriologist and Assistant Bacteriologist, Louisiana Sugar Experiment Station 



Molds have long been known to produce intracellular and extracel- 

 lular enzyms, but so far as we have been able to ascertain no study has 

 been made of the enzyms contained in the spores of these organisms. 

 This is indeed noteworthy, since the valuable contributions of Duclaux, 

 Fembach, and others mentioned by Effront^ and Dox^ have apparently 

 been concerned solely with the enzymic activities of molds in the 

 mycelial stage. It has been shown by us in another connection* that 

 the deterioration of sugar occurred where mold mycelia developed and 

 in certain instances where spores alone were present. This phenomenon 

 gave rise to the query, Do mold spores contain enzyms? This is the 

 concern of the present investigation. It is our purpose to limit the 

 scope of this article to the invertase activity of the spores of Aspergillus 

 niger (Aspergillus sydowi [Bain, and S.]), Aspergillus flavus, and 

 Penicillium expansum, because of the economic significance of this enzym 

 and the wide distribution of these molds. 



METHOD OF PROCEDURE 



A large number of Petri dishes containing Kopeloff 's agar* were 

 seeded with a single pure, bacteria-free culture of the desired mold and 

 incubated at 35° C. for six days. A small amount of sterile distilled 

 water was introduced into each plate, shaken slightly, and then poured 

 on sterilized filter paper (Whatman No. 4). The mycelium was then 

 washed through the filter paper with sterile distilled water, and the 

 spores were separated by a combination of filtration and flotation. 

 This process was continued long enough to accumulate the desired quan- 

 tity of spores. The spores were then rinsed off the filter paper into a 

 sterile Erlenmeyer flask of 300-cc. capacity. To 250 cc. of this spore 

 suspension and an equal volume of sterile distilled water, i cc. of c. p. 

 chloroform was added. 



1 The authors are indebted to Director W. R. Dodson, Assistant Director W. G. Taggart, Dr. F. W. 

 Zerban, and Mr. E. C. Freeland for their kind interest and assistance, and to Mr. W. L. Owen for reading 

 the manuscript. 



2 Effront, Jean, enzymes and their applications . . . English translation by Samuel C. Prescott. 

 322 p. New York, 1902. 



' DOX, A. W. THE INTRACELLULAR ENZYMES OP PENICILLIUM AND ASPERGILLUS ... U. S. Dept. 



Agr. Bur. Anim. Indus. Bui. 120, 70 p. 1910. 



* Kopeloff, Nicholas, and Kopeloff, Lillian, the deterioration of cane sugar by fungi. La. 

 Agr. Exp. Sta. Bui. 166. 



Journal of Agricultural Research, Vol. XVIII, No. 4 



Washington, D. C. Nov. 15. 1919 



cv Key No. La.-i 



(195) 



