NON-OXIDATIVE ENZYMES OF SPORE EXTRACTS 



100'- 



80 



60 



^ 



40 



20 



10 



30 



20 

 MINUTES 

 Fig. 12. Pyrophosphatase of extracts of resting spores (RS) and of germi- 

 nated spores (GS). Spores ground in Mickle Disintegrator. RS ground 

 after heating at 50°C for 15 min. GS prepared by incubation of RS with 

 0.025 M glucose for 1.5 hr. at 30°C (81% germination) followed by grinding. 

 All mixtures contained 0.25 ml extract, 5 x 10~*M manganous sulfate, 10~^ 

 M Na4P207, and veronal buffer at pH 7.0 in a total volume of 2.0 ml. Re- 

 action stopped at indicated time by addition of 0.2 ml 100% trichloroacetic 

 acid. 100% hydrolysis is calculated to yield 124.1 /xg P. 



be of significance that the extract of the so-called germinated spores was 

 actually derived from spores which had germinated 81%, or to put it another 

 way, 19% of whose spores had not germinated. Could this signify that only 

 the resting spores had appreciable amounts of pyrophosphatase, and that 

 the small amount of activity evidenced by the extracts of germinated spores 

 was due to residual resting spores? This appears to be quite an unusual 

 situation, and one which I believe is worth further investigation. 



The spore coats of B. megaterium are rich in phosphorus which is acid 

 and alkali insoluble. Fitz- James (1955) estimates that this P fraction is 

 about 60% of the total P of B. megaterium spores. It is possible that this 

 insoluble residue forms a lattice work making the spore coat impermeable to 

 nutrients. The breakdown, or partial breakdown, of this lattice work through 

 the mediation of the manganese-activated pyrophosphatase would permit 

 nutrients to enter the spore and to participate in the biochemical events 

 necessary for germination of the spore. 



T realize that I have neglected some important non-oxidative enzymes. 

 Perhaps Dr. Krask will touch on these. I hope that when the time for gen- 



