NON-OXIDATIVE ENZYMES OF SPORE EXTRACTS 



125 



REACTION 

 MIXTURE 



DAYS 



2 3 



o 



o> o o 



o o o 



o o 



o o o 



o o 



EA-f 

 X + Mnl AX 



Qo oQQ 



o o o o o 



Fig. 4. Tracing of paper partition chromatogram showing hydrolysis of 

 egg albumin by spore extract. EA = egg albumin (1.0 per cent): other 

 conditions as for Fig. 3. 



two days; but with manganese, definite signs of liberation of amino acids 

 were observable in this time. 



(2) A colorimetric method based on the reaction of free amino groups 

 with ninhydrin. The proteinaceous substrate-enzyme mixture was incubated, 

 treated with ninhydrin, and the resulting color was read in a Klett colorim- 

 eter. By this method, too. a breakdown of the protein substrate is evident, and 

 manganese is decidedly stimulatory to this breakdown (Fig. 5). The ques- 

 tion arises here as to whether manganese is an artifact. That is, is it possi- 



Table III 

 Eflfect of manganese on ninhydrin reaction 



"Amino Acid' 



Manganese 



Klett^ 



Sodium Glutamate 

 Bacto Peptone 

 DL-<Alanine 



+ 

 + 

 + 



188 

 185 

 22 

 25 

 104 

 112 



The Klett reading indicates the intensity of the ninhydrin color 



