NON-OXIDATIVK KNZVMKS OF SI'OKK KXTKVCTS 1:^7 



length. The latter activities occurred at pH 0.8 while that of specific GTF 

 activity occurred at pH 6.5. 



(3) (X)— Transferase (XTF) 



Adenosine 

 (X) + NH.OH — (Xl- Hydroxamic acid (XHA) + NH, 



In our course of study on GTF it was noted that a reaction occurred w ith 

 hydroxylamine in the absence of glutamine. This unidentified transferase 

 (XTF) occurs only in the spore homogenate or debris and reacts with a 

 bound non-dialyzable substrate in the spore. Further, the reaction required 

 no metal ions and was stimulated by ATP, ADP, AMP and adenosine — 

 optimal stimulation occurred with adenosine. ATP was the least effective 

 of the nucleotides while adenine was completely inactive. The pH range 

 of the enzyme is broad with an apparent acid optimum. The stimulation by 

 adenosine and activity with a substrate proper to the spore itself suggests this 

 enzyme is functional in germination. 



(4) Acetokinase-like enzyme 



Mg 

 Acetate + NH^OH + ATP ^ Acetohydroxamic acid ( AcHA) 



The fourth enzyme studied has been tentatively called an acetokinase-like 

 enzyme but is in all probability an aceto-CoA-kinase as described by Jones 

 and Lipmann (1955). The reaction of acetate with hydroxylamine to form 

 acetohydroxamic acid (AcHA) can only occur through the formation of 

 acetyl phosphate or acetyl-CoA which then react with hydroxlamine. This 

 reaction occurs in spore homogenates and vegetative cells and requires ATP 

 and Mg+ +. The reaction appears to be independent of added Co A, but in the 

 light of Murrell's studies (1955) demonstrating CoA in spores and our 

 results of extract studies it is probably a CoA mediated reaction. 



Preliminary studies also show that the spore homogenates contain 

 ATP-ase. 



While the studies of these enzymes were concerned primarily with a char- 

 acterization of their properties, the following tables demonstrate their occur- 

 rence, their relative activity, and the effect of heat shock on this activity. 

 All comparisons are on the basis of 20 mg dry weight or its equivalent. 



Table I demonstrates the occurrence and relative activities of the enzymes. 

 The activity associated with the intact spores may be questionable since heat 

 shock for 30 minutes had no effect, although, as will be demonstrated, heat 

 shock prior to disruption has a marked effect on the enzymes. The activity of 

 all the enzymes becomes manifest upon disruption for one hour. The uniden- 

 tified transferase appears only in the disrupted spores and is absent from a 

 ten hour intact vegetative cell suspension. 



