OXIDATIVE ENZYMES OF BACTERIAL SPOltE EXTRACTS 



145 



Fig. 1. The activation of glucose 

 oxidation by heat. Clean spores of 

 B. cereus var. terminalis were heat 

 shocked at 65C. cooled and tested 

 for respiratory activity by the two 

 cup method. Warburg flask contents: 

 Main compartment, 30 mg spores in 1 

 ml 0.067 M phosphate buffer, pH 

 7.2; center well. 0.2 ml 30 per cent 

 KOH where appropriate; side arm, 1 

 mg glucose. Total volume 2.1 ml. Re- 

 action run at 30C under atmospher- 

 ic oxygen. 



Minuses 



Fig. 2. Oxidative capacities of ac- 

 tivated spores. See legend Fig. 1 for 

 details. Substrates were added at a 

 level of 5/i,mole/ml. 



cereus var. terminalis is shown in Fig. 1. The oxidation capacities of acti- 

 vated spores are shown in Fig. 2. In addition to glucose, these spores rapidly 

 oxidize 2-keto gluconate (2KG), pyruvate, and gluconate. Glucose-6-phos- 

 phate (G-6-P), fructose-6-phosphate (F-6-P), fructose (F), hexose diphos- 

 phate ( HDP I . and acetate are not attacked. Thus, activated spores are ex- 

 cellent material with which to elucidate the pathways of glucose metabolism 

 which furnish biologically useful energy and intermediates for biosynthesis 

 during germination. 



Since the approach to an understanding of the metabolic pathways oper- 

 ative in activated spores required measurements of single-step reactions of 

 a metabolic chain, experiments were performed with crude spore extracts 

 prepared by the following procedure: Twenty grams of superbrite glas? 

 beads were suspended in 20 ml of 0.067M phosphate buffer at pH 7.4 with 

 two grams of dry spores. The mixture was ground in a Waring blender at 



