Biochemical characterization of a semi-purified aspartic protease from sea catfish Bagre panamensis with milk-clotting activity.

Pepsin from stomach of Bagre panamensis was semi-purified and biochemically characterized. The acid proteolytic activity and purification fold were 3875 U/mg protein and 91.85, respectively, after purification process. The optimum pH and temperature for semi-purified protease were 2-3 and 65 °C, respectively. The enzyme activity was stable after heating proteases at 50 °C for 120 min, but only 30% residual activity was detected after heating at 65 °C for 30 min. SDS-PAGE analysis showed two proteins bands after dialysis (26.1 and 38.6 kDa). Only the band of 38.6 kDa had proteolytic activity, which was inhibited using pepstatin A. Organic solvents, surfactants and reducing agents affect the proteolytic activity at different extent; however, metal ions or EDTA have no impact on protease activity. The semi-purified protease exhibited milk coagulant activity, with a maximum activity at 45 °C. The obtained results highlight the potential biotechnological use of B. panamensis pepsin.

Effect of fishmeal replacement by soy protein concentrate with taurine supplementation on growth performance, hematological and biochemical status, and liver histology of totoaba juveniles (Totoaba macdonaldi).

The effect of dietary inclusion of soy protein concentrate (SPC) and simultaneous supplementation with taurine on the growth, hematology, blood biochemistry, and liver histology of totoaba (Totoaba macdonaldi) juveniles was assessed. Four isoproteic and isolipidic diets were formulated containing either 30 or 60% of SPC (diets S30 and S60), supplemented or not with 1% of taurine (diets S30T and S60T). A fishmeal-based diet formulated for totoaba nutritional requirements, without SPC and taurine supplementation, was used as a reference diet. Triplicate groups of 32 totoaba juveniles (average body weight 7.5 ± 0.6 g) were fed these diets for 45 days. Results showed that growth performance in fish fed S30, S30T, and S60T was similar to fish fed the reference diet. Red blood cells and hematocrit in fish fed with supplemented taurine in both levels of SPC (S30T and S60T) were similar to the fish fed the RD; the addition of taurine improved the state of hydration of totoaba. Plasmatic hemoglobin in fish fed the lower SPC level was similar to fish fed the RD. The mean corpuscular hemoglobin concentration in fish fed S30T was similar to fish fed the RD, taurine supplementation prevented the development of hypochromic anemia in this group of fish. Plasmatic albumin in fish fed S30 was similar to fish fed the RD. Plasmatic total protein and globulin concentration increased and AL:GLB (albumin:globulin ratio) decreased in fish fed the SPC-based diets despite taurine supplementation. The protein profile showed that taurine supplementation did not prevent a possible inflammatory process (increased globulins, decreased AL:GLB) in juvenile totoaba fed both levels of SPC. Glucose concentration was similar in fish fed S30, S30T, and S60T. The histological hepatic index was highest in fish fed S60. These results suggest that with an appropriate nutritional level, taurine may play an important modulatory role in the hematology and blood biochemistry status in totoaba fed SPC-based diets, contributing to the enhancement of an overall healthy growth performance.

Effect of fishmeal replacement by soy protein concentrate with taurine supplementation on hepatic intermediary metabolism and antioxidant status of totoaba juveniles (Totoaba macdonaldi).

The effect of dietary incorporation of soy protein concentrate (SPC) and the concomitant supplementation with taurine on hepatic intermediary metabolism and antioxidant status of totoaba (Totoaba macdonaldi) juveniles was assessed. Four isoproteic and isolipidic diets were formulated containing either 30 or 60% of SPC (diets SP30 and SP60), supplemented or not with 1% of taurine (diets SP30T and SP60T). A fish meal (FM) based diet, without SPC and taurine supplementation, was used as a control. Triplicate groups of 32 totoaba juveniles (average body mass=7.5g) were fed these diets over 45days. Results revealed that dietary FM replacement by SPC depressed the overall intermediary metabolism. Activity of key enzymes of amino acid catabolism and gluconeogenesis was significantly reduced and a trend to reduce glycolysis and glucose-6-phosphate dehydrogenase activity was observed. The incorporation of the highest level of SPC also significantly increased hepatic lipid peroxidation and the activity of superoxide dismutase. Concomitant taurine supplementation restored the activity of amino acid catabolic and gluconeogenic enzymes and hexokinase to levels similar of those of the control diet. Taurine supplementation also led to a significant increase of glucose-6-phosphate dehydrogenase and catalase activity, as well as to a significant reduction of liver lipid peroxidation. These results suggest that taurine may play an important metabolic modulation action on totoaba fed SPC based diets, contributing to the enhancement of the overall metabolism and to the reduction of liver oxidative damage.