156 harlyn halvorson 



Table 111 

 Accumulation of pyruvate during the deamination of alanine by 

 intact spores in the presence of an inhibitor of pyruvate oxidation 



The incubation mixture contained 100 mg of spores, thiamine at a final 

 concentration of 6.6xlO""'M, W-1455 at a final concentration of 5xlO-^M and 

 0.067M phosphate buffer pH 7.0 to a final volume of 1.5 ml. After 60 min. 

 incubation. NH3 was determined by distillation and pyruvate after centrif- 

 ugation by the method of Friedman and Haugen ( 1942 ) . Germination was 

 measured by the uptake of an aqueous methylene blue. 



I B,jP I for the formation of NH3 and pyruvate from D- or L-alanine, Since 

 these extracts contain an active alanine racemase (Stewart and Halvorson, 

 1953), the deamination of both D- and L-alanine may represent a non-spe- 

 cificity on the part of the deaminase system or coupled reaction with alanine 

 racemase. The presence of catalase in these extracts ( Lawrence and Hal- 

 vorson, 1954) precluded an examination for the H2O2 observed by Falcone, 

 Presumably these two reactions are similar, 



NH3, H20i>, and pyruvate can be directly formed from alanine by the 

 action of either D- or L-amino acid oxidase (Meister, 1955). Since these 

 require either flavin adenine dinucleotide or riboflavin phosphate but not 

 B(iP as cofactors, they differ from the requirements of the above system. 

 Alternatively the observed reaction might be mediated by a B(;P activated 

 glutamic-alanine transaminase (Meister, 19551 followed by a deamination 

 of glutamate. Although vegetative cells of the Bacillaceae contain active glu- 

 tamic-aspartic ( Hardwick and Foster, 1953; Keynon et al, 1954; Levinson 

 and Sevag, 1954; Meister, 1955) and glutamic-alanine ( Harwick and Fos- 

 ter, 1953; Levinson and Sevag, 1954; Meister, 1955 1 transaminases, only 

 glutamic-aspartic transaminase has been reported in spore extracts of Bacil- 

 lus megatherium (Levinson and Sevag, 1954). Hardwick and Foster (1953) 

 were unable to detect glutamic-alanine transaminase activity in spore ex- 

 tracts of B. mycoides. We have confirmed this observation employing ex- 

 tracts of B. cereus var. terminalis spores which actively deaminate alanine. 

 This unique reaction can be fornmlated as follows: (Fig, 14). 



