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STUDIES ON CELL METABOLISM AND CELL DIVISION : VII. OBSERVATIONS ON THE AMOUNT AND POSSIBLE FUNCTION OF DIPHOSPHOTHIAMINE (COCARBOXYLASE) IN EGGS OF ARBACIA PUNCTULATA
Authors:M E Krahl  Bernhard J Jandorf  G H A Clowes
Institution:From the Lilly Research Laboratories, Indianapolis, Indiana, and the Marine Biological Laboratory, Woods Hole, Massachusetts
Abstract:1. Methods suitable for the determination of diphosphothiamine (cocarboxylase) in eggs of Arbacia punctulata have been developed. Quantitative extraction of the cocarboxylase was effected by combining the use of thiamine hydrochloride in the extraction fluid with critical adjustment of the pH of extraction to pH 6.3–6.7. 2. The unfertilized eggs were found to contain the equivalent of 2 to 3 micrograms of natural yeast cocarboxylase per gm. of wet eggs; the cocarboxylase content of the 30 minute and 10 hour fertilized eggs was somewhat less (Table III). 3. In preliminary experiments, Arbacia egg cytolysates were found to cause pyruvic acid to disappear. The rate of such disappearance was apparently greater under aerobic than under anaerobic conditions; it was also greater for cytolysates from fertilized eggs than for cytolysates from unfertilized eggs (Table IV).
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