The Acorn Worm Balanoglossus biminiensis

Dure and Cormier (1961) demonstrated a luciferin-luciferase reaction for the first time in the extracts of the acorn worm Balanoglossus biminiensis, and also discovered that the luminescence reaction is stimulated by H2O2, of which the details are described below. Recently, Kanakubo and Isobe (2005) reported the chemical structure of a probable luciferin of another acorn worm Ptychodera flava. 

According to Dure and Cormier (1961, 1963) and Cormier and Dure (1963), they made the preparations of luciferase and luciferin from Balanoglossus biminiensis, collected on Sapelo Island, Georgia, and investigated the luciferin-luciferase reaction, as summarized below. 

Preparation of luciferase. Organisms were freeze-dried, powdered, and washed with ethyl acetate to destroy the majority of catalase activity. The washed residue was extracted with 50 mM potassium phosphate buffer, pH 6. The extract was fractionated by ammonium 

Preparation of Balanoglossus luciferin. The residue of the first pH 6 extraction above was re-extracted with 50 mM potassium phosphate buffer, pH 8. After centrifugation, the supernatant was used as the standard luciferin preparation. Luciferin was highly labile and easily inactivated at an extreme pH, by heat, and also by freezing and thawing. The instability resembled that of certain proteins. 

Cormier and Dure (1963) found another type of luciferin and called it protein-free luciferin. Protein-free luciferin was found in the vapor condensate of freeze-drying whole animals, and also in the 3 5-56 % ammonium sulfate fraction of the crude extract noted above. The protein-free luciferin behaved like an aromatic or heterocyclic compound and it was strongly adsorbed onto Sephadex and other chromatography media, requiring a considerable amount of solvent to elute it. The luminescence reaction of protein-free luciferin in the presence of luciferase required a 500-times higher concentration of H2O2 compared with the standard luciferin preparation. Both types of the luciferin preparation had a strong odor of iodoform. 

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Fig. 10.4.1 Light emission profile of the luminescence reaction of the acorn worm Balanoglossus biminiensi, when H2O2 is injected into a mixture of the luciferin and luciferase. From Dure and Cormier, 1961, with permission from the American Society for Biochemistry and Molecular Biology.

Isolated from the acorn worm Balanoglossus biminiensis, likely used as a hormone... 

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