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Anionic trypsin from albacore tuna spleen was purified by chromatographic separations on Q-Sepharose, Superdex 75 and Arginine Sepharose 4B. The trypsin migrated as single bands in both SDS-PAGE and native-PAGE. The molecular weight of purified trypsin was estimated to be 30 kDa using SDS-PAGE. The enzyme exhibited maximal activity at pH 9.0 and 55 °C for hydrolysis of Boc-Val-Pro-Arg-MCA. pH and temperature stabilities of the trypsin were well maintained in the pH range of 6-11 and over a temperature range from 20 up to 50 °C. The enzyme was effectively inhibited by soybean trypsin inhibitor, N‑tosyl‑l‑phenyl‑alanine chloromethyl ketone (TLCK) and Pefabloc SC. The N-terminal amino acid sequence of 20 residues of the purified enzyme was IVGGYECQAHSQPHQVSLNA, which is highly homologous to other fish trypsins. The k/K of the enzyme for Boc-Val-Pro-Arg-MCA was 2.60 ± 0.07 s mM. Purified trypsin also hydrolysed fish muscle proteins, suggesting its effectiveness in degradation of food proteins.
This article was published in the following journal.
Name: International journal of biological macromolecules
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A congenital or acquired condition in which the SPLEEN is not in its normal anatomical position but moves about in the ABDOMEN. This is due to laxity or absence of suspensory ligaments which normally provide peritoneal attachments to keep the SPLEEN in a fixed position. Clinical symptoms include ABDOMINAL PAIN, splenic torsion and ISCHEMIA.
A pancreatic trypsin inhibitor common to all mammals. It is secreted with the zymogens into the pancreatic juice. It is a protein composed of 56 amino acid residues and is different in amino acid composition and physiological activity from the Kunitz bovine pancreatic trypsin inhibitor (APROTININ).
Serine proteinase inhibitors which inhibit trypsin. They may be endogenous or exogenous compounds.
A high-molecular-weight protein (approximately 22,500) containing 198 amino acid residues. It is a strong inhibitor of trypsin and human plasmin.
Vein formed by the union (at the hilus of the spleen) of several small veins from the stomach, pancreas, spleen and mesentery.
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