Puerta-Martínez, Francisco
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Publication Fluorescent based assessment of trypsin activity vs total proteases in the queen triggerfish, Balistes vetula(2007) Puerta-Martínez, Francisco; Uscian, John M.; College of Arts and Sciences - Sciences; Williams, Lucy B.; Montalvo, Rafael; Kubaryk, John; Department of Biology; Grove, KurtAmino acids that comprise proteins constitute essential nutrients for any animal organism. Indeed they are directly related to its growth and maintenance. Protein assimilation depends on the catalytic properties of proteases in the alimentary canal. Such protein-degredative enzymes include trypsin, chymotrypsin and elastase, with trypsin being the most important and abundant. The main objective of this research project was to use a highly sensitive fluorometric technique to assess both total protease and trypsin activities from intestines of the queen triggerfish, Balistes vetula, and determine if these were affected by fish size. To this end, 32 west coast and 4 east coast intestinal samples were obtained. Each sample contained from 2 – 10 total intestines. The weight (wet weight) was obtained and recorded for each sample. Samples were then individually homogenized in distilled water to produce a supernatant. Filtered supernatant was assayed for the following three properties: 1) protein content; 2) total protease activity; 3) trypsin activity. The remaining supernatant was pooled and subjected to size exclusion chromatographic procedure in order to obtain a partial purification of proteins conferring trypsin activity. The kinetic analysis of this trypsin activity was assessed as per the following four variables: 1) protein concentration; 2) pH; 3) temperature; and 4) presence of soybean trypsin inhibitor. The data are considered with respect to both the utility of the fluorometric technique in this analysis and the significance of trypsin with respect to the biochemistry/physiology of the queen triggerfish.