F. I. TSUJI, A. M. CHASE AND E. N. HARVEY 153 



fluorescence in ultraviolet light. The paper areas on both sides of the 

 luciferin region were also hydrolyzed, both hydrolyzates giving a very 

 strong ninhydrin test. When ahquots of the hydrolyzates from paper 

 chromatography and from paper electrophoresis were repeatedly made 

 alkaline and evacuated each time to dryness in a desiccator, they 

 still gave strong positive ninhydrin tests, indicating that the ninhydrin- 

 positive material was not a volatile substance like ammonia. From the 

 ninhydrin color value, 25% of the luciferin hydrolyzate from paper 

 chromatography could be expressed in terms of equivalence of leucine. 

 Following these tests, the remainder of the hydrolyzate of luciferin 

 from the paper chromatography isolation was analyzed for amino 

 acids in the long Dowex 50 column according to the method of Moore 

 and Stein (1951). The following compounds, listed in approximate 

 order of abundance, were found to be present in the hydrolyzate: 

 either aspartic acid, threonine, or serine; ammonia, arginine, leucine, 

 isoleucine, glutamic acid, alanine, vahne, sarcosine, lysine, taurine, and 

 methionine. Of these compounds ammonia and either aspartic acid, 

 threonine or serine were found in very high concentration, although 

 ammonia, arginine, and lysine may be present in higher concentration 

 than indicated. The presence of proHne and histidine was uncertain, 

 whereas tyrosine and phenylalanine were absent. There is no doubt 

 but that doubly cycled luciferin contains amino acids. The positive 

 ninhydrin reaction given by hydrolyzates from other regions of the 

 paper chromatogram may have come from oxidized luciferin or poly- 

 peptide impurities. 



References 



Anderson, R. S. 1933. The chemistry of bioluminescence I. Quantitative 

 determination of luciferin. /. Cellular and Comp. Physiol, 3, 45-59. 



Anderson, R. S. 1935. Studies on bioluminescence II. The partial purifica- 

 tion of Cypridina luciferin. /. Gen. Physiol, 19, 301-305. 



Anderson, R. S. 1936. Chemical studies on bioluminescence III. The re- 

 versible reaction of Cypridina luciferin with oxidizing agents and its 

 relation to the luminescent reaction. /. Cellular and Camp. Physiol, 8, 

 261-76. 



Anderson, R. S. 1937. Chemical studies on bioluminescence IV. Salt effects 



