Developing a Fish Meat‐binding Agent: Purification of Salmon Thrombin

Abstract

Thrombin from Atlantic salmon (Salmo salar) was purified and characterized as a potential new binding agent for the food industry. Purification was performed avoiding inhibitors, using BaSO 4 adsorption and heparin-Sepharose affinity chromatography. Prothrombin activation was performed using a mixture of eggs and gills from salmon. Optimized conditions for the adsorption, elution, and the activation step are presented. The purified thrombin clotted bovine fibrinogen with a specific activity of 1423 U/mg. Sequence data are presented and compared with other species. This method of nontoxic activation and purification will allow salmon thrombin to be used in the food industry.

Sample displacement chromatography of Atlantic Salmon (Salmo salar) thrombin

“…2. We have recently purified salmon thrombin from plasma [3] where the thrombin activity yield in the activated BaSO 4 eluate was 22%. In this study, the activated BaSO 4 eluate was further purified and the resulting thrombin activity yield was as much as 71.7%.…”

“…The prothrombin isolation and activation procedure is described in detail by Manseth et al [3]. Briefly, blood was collected from Atlantic salmon and added sodium oxalate to a final concentration of 0.01 M. Plasma was separated by centrifugation, and the prothrombin was adsorbed onto BaSO 4 and eluted using 0.16 M sodium citrate.…”

“…Salmon thrombin activity was calibrated at 26 jC [2] by using the method described by Fenton et al [11], and routinely measured using the chromogenic substrate S-2238 [12] as described by Manseth et al [3]. …”

“…The idea of non-homogeneous application is to reduce the probability of the lower affinity components to compete with the components with higher affinity during the displacement. The advantages of using such an approach became clear during our work with salmon thrombin [3], and we wanted to study this method in more detail.…”

“…Salmon thrombin has been suggested as alternative to mammalian thrombins for the use as binding agent in surgical 0165 applications [1,2]. We have recently presented a non-toxic purification method for Atlantic salmon (Salmo salar) thrombin for the potential use as a component in a fish meat-binding agent [3], by using affinity chromatography (Heparin Sepharose) and ion exchange chromatography (SP Sepharose).…”