Rivera-Santos, Mishelle

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Purification and characterization of trypsin from intestinal and pyloric caecal tissues from the silk snapper, Lutjanus vivanus (Cuvier, 1828)
(2003) Rivera-Santos, Mishelle; Uscian, John M.; College of Arts and Sciences - Sciences; Navas, Vivian; Bunkley-Williams, Lucy; Department of Biology; Kubaryk, John
In an effort to better understand the chemical digestion of proteins in the silk snapper, Lutjanus vivanus (Cuvier, 1828), and thereby obtain a deeper understanding of how this fish meets its dietary needs for amino acids through its trophic environment, trypsin was purified 10-fold from intestinal and pyloric caecal tissues by sequential application of ammonium sulfate fractionation, size exclusion chromatography, and affinity chromatography. Standard kinetic characteristics of this enzyme were determined. The enzyme displayed optimal activity at pH 8 and 60oC. It was completely or nearly completely inhibited by soybean trypsin inhibitor (SBTI) at a concentration of 0.1% or higher. SDS-PAGE analysis of the purified enzyme revealed a single band with an estimated molecular weight of 26.1 kDa.