OXIDATIVE ENZYMES OF BACTERIAL SPORE EXTRACTS 



155 



MINUTES 



Fig. 13. Oxidation of various substrates by particles from spore extracts. 

 A dialyzed spore extract (see Fig. 3) was centrifuged at 140,000 times grav- 

 ity for 1 hour. The pellet was reconcentrated at one-eighth its original vol- 

 ume in 0.067 M phosphate buffer, pH 7.1. The Warburg flask contents: side 

 arm, 25/x moles of substrate and O.OS/x mole of OAA; center well, 0.2 ml 30 

 per cent KOH ; main compartment, 1 ml enzyme preparation, 8 x lO'^ M 

 MnClo. 0.2 ml of 0.05 M ATP. 5x10=* M DPN, 10"^ M cocarboxylase, and 

 sufficient phosphate buffer to a final cell volume of 2.0 ml. The gas phase 

 was air and the temperature 30C. 



B. cereus var. terminalis actively deaminate alanine but not glycine or pheny- 

 lalanine (Table III). When the pyruvate oxidation present in these spores 

 is suppressed by W-1435, quantitative recoveries of pyruvate are obtained 

 from alanine. W-1435 also inhibits spore germination in the presence of 

 alanine. Thiamine, which reverses the inhibition of pyruvate oxidation by 

 W-1435 (Edwards et al, 1956), restores partial germination in the presence 

 of alanine. Dialyzed extracts of these spores require pyridoxal phosphate 



