Characterization of digestive enzymes during early ontogeny of white Snook (Centropomus viridis).

Abstract

The white snook (Centropomus viridis) is a species with great aquaculture potential, therefore, the present study aims two objectives; 1) to characterize digestive enzymatic activity during larval development and; 2) characterize the diversity and functionality of digestive enzymes in an early juvenile stage of C. viridis. On first feeding, larvae were 2.75 ± 0.09 mm total length and grew to 24.35 ± 4.35 mm after 42 days. The specific activity of acid proteases was detected with low activity from day 1 (Days after hatching, DAH) to day 21, and increased at 23 DAH, reaching maximum activity at 40 DAH. Meanwhile, digestive enzymatic activity of alkaline proteases was observed from hatching to day 40. From hatching until 40 DAH, the presence of alkaline proteases, trypsin, chymotrypsin, leucine aminopeptidase, α-amylase and lipase were detected with different activity patterns. Total and specific activity of digestive enzymes on juveniles showed higher activity of acid than alkaline proteases, while α-amylase had a higher activity than lipases. The maximum activity of acid proteases was observed at 40 °C and at pH 2.0, while digestive alkaline proteases showed maximum activity at 60 °C. α-amylases and lipases showed their maximum activities at 50 °C. The maximum alkaline protease and α-amylase activities were found at pH 7 to 9. The enzymatic activity of juvenile's acid proteases was highly inhibited by Pepstatin A (96%), however the activity of alkaline proteases incubated with PHEN, TLCK, SBTI, PMFS, and OVO showed reductions of 51.29, 30.16, 27.98, 27.26, and 24.68% respectively. Lipase activity was totally inhibited with Ebelactone B, and highly inhibited by Ebelactone A, Orlistat, 1% SDS, and PMFS (96.12, 77.16, 64.22, and 51.29%, respectively). Seven bands (102.6, 65.7, 58.9, 40.6, 27.4, 21.5, and 15.8 kDa) resolved by protein electrophoresis were detected with alkaline protease activity, identified as serine proteases and metalloproteases, and three bands (57, 43.9, and 32.6 kDa) with lipase-like activity were also identified. These results show that C. viridis ontogeny entails an indirect development of its digestive system with a functional stomach on day 23, and characterized by increases in pepsin activity. Indeed, white snook ontogeny shows a similar developmental trajectory to other carnivorous marine fish, showing a diversity of alkaline proteases that work in a wide range of pH and temperatures. Thus, the identification of these enzymes provides advantages to formulate white snook aqua-feeds.