The effect of heating conditions on polyphenol oxidase, proteases and melanosis in pre-cooked Pacific white shrimp during refrigerated storage

Pre-cooked Pacific white shrimp (Litopenaeus vannamei) is an important shrimp product. However melanosis, especially in the cephalothorax including carapace and internal organs, is more likely caused by the remaining polyphenol oxidase (PPO) after pre-cooking. Thus, PPO from carapace and proteases from hepatopancreas of Pacific white shrimp were characterised and the remaining activities of both enzymes were monitored in pre-cooked shrimp during storage at 4 °C. Based on activity staining using L-β-(3,4 dihydroxylphenyl) alanine as a substrate, PPO consisted of two isoforms with apparent molecular weight of 210 and 220 kDa. No difference in activity band was observed when analysed under reducing and non-reducing condition. Proteases from hepatopancreas were able to activate PPO to some degree. For the in vitro study, both enzymes were quite stable when heated at temperature up to 70 °C but the loss in activities increased with increasing heating time (0-120 s). When Pacific white shrimp were pre-cooked to obtain different core temperatures (50-90 °C), different PPO and protease activities were retained. Higher core temperatures were associated with lower PPO and protease activities, but higher cooking loss. When the shrimp were pre-cooked at 80 °C, the residual PPO and protease activities were 3.9% and 5.4%, respectively and cooking yield of 95.6% was obtained. The resulting pre-cooked shrimp possessed lower melanosis score during 7 days of storage at 4 °C. Thus, pre-cooking of shrimp to obtain a core temperature of 80 °C, with a holding time of 30 s, could prevent the severe cooking loss and lower melanosis during subsequent storage.