164 JOAN F. POWELL 



substrate, a lO^Yml spore suspension in M/20 phosphate pH 7.3 was dis- 

 integrated, and 1 ml of the suspension (= 5.5 mg dry wt) added to 5 ml 

 of a 6 X 10"^M solution of reduced ( Na2S204 ) cytochrome C in M/20 phos- 

 phate pH 7.3. After 15 min. at room temperature there was a marked color 

 change from pink to orange and the absorption band at 550 xufi had almost 

 disappeared. During this time there was no appreciable change in a control 

 containing no spores. After centrifuging, the activity of the supernatant 

 was considerably less active than that of the suspension. The supernatant 

 was added to an equal volume of 4 x 10~"' M reduced cytochrome C and 

 the decrease in absorption at 550 m/x followed in a Unicam spectrophoto- 

 meter. The oxidation was slow (50% substrate oxidized in 20 min.) but 

 definite. In a control containing supernatant heated at 100° for 10 min., 

 oxidation was extremely slow, i.e., less than 50% in 60 min. The more 

 active uncentrifuged suspension could not be used for these measurements, 

 being far too turbid. Disintegrated germinated spores showed approximately 

 the same activity as disintegrated resting spores whereas disintegrated 

 vegetative cells appeared, on a dry wt. basis, to be roughly ten times as 

 active in the oxidation of reduced cytochrome C. 



Comparison of the Coenzyme I content of extracts from resting 

 and germinated spores of B. siibtilis (laboratory strain) 



The method of assay described by Gingrich and Schlenk ( 194 1, J. Bact. 

 47: 535) was used with H. parainfluenzoe NCCT 4101 as test organism. 

 The standard Col preparation was supplied bv Schwarz Laboratories as 

 50% pure Col. On this basis, it permitted barely visible growth at 

 0.0017 y Col/ml. The amount of growth increased linearly in the range 

 used, i.e., 0.01-0.05y/ml. Resting and germinated (L-alanine) spores of 

 B. subtjlis were disintegrated, centrifuged and the extract sterilized by 

 filtration. The sterile extract was added to the basal medium. There ap- 

 peared to be no increased svnthesis of Col during germination: in fact, 

 in duplicate experiments, the Col content of extracts from germinated spores 

 was lower than that from resting spores. 



Col content (y) of extract from 10^*' spores 



Resting Germinated 



(5.5 mg dry wt.) (3.9 mg dry wt.) 



Exp. I 0.13 0.08 



Exp. II 0.14 0.10 



