Molecular, Biochemical, and Dietary Regulation Features of alpha-Amylase in a Carnivorous Crustacean, the Spiny Lobster Panulirus argus

作者:Rodriguez Viera Leandro*; Perera Erick*; Antonio Martos Sitcha Juan; Perdomo Morales Rolando; Casuso Antonio; Montero Alejo Vivian; Garcia Galano Tsai; Martinez Rodriguez Gonzalo; Miguel Mancera Juan
来源:PLos One, 2016, 11(7): e0158919.
DOI:10.1371/journal.pone.0158919

摘要

Alpha-amylases are ubiquitously distributed throughout microbials, plants and animals. It is widely accepted that omnivorous crustaceans have higher alpha-amylase activity and number of isoforms than carnivorous, but contradictory results have been obtained in some species, and carnivorous crustaceans have been less studied. In addition, the physiological meaning of alpha-amylase polymorphism in crustaceans is not well understood. In this work we studied alpha-amylase in a carnivorous lobster at the gene, transcript, and protein levels. It was showed that alpha-amylase isoenzyme composition (i.e., phenotype) in lobster determines carbohydrate digestion efficiency. Most frequent alpha-amylase phenotype has the lowest digestion efficiency, suggesting this is a favoured trait. We revealed that gene and intron loss have occurred in lobster alpha-amylase, thus lobsters express a single 1830 bp cDNA encoding a highly conserved protein with 513 amino acids. This protein gives rise to two isoenzymes in some individuals by glycosylation but not by limited proteolysis. Only the glycosylated isoenzyme could be purified by chromatography, with biochemical features similar to other animal amylases. High carbohydrate content in diet down-regulates a-amylase gene expression in lobster. However, high alpha-amylase activity occurs in lobster gastric juice irrespective of diet and was proposed to function as an early sensor of the carbohydrate content of diet to regulate further gene expression. We concluded that gene/isoenzyme simplicity, post-translational modifications and low Km, coupled with a tight regulation of gene expression, have arose during evolution of alpha-amylase in the carnivorous lobster to control excessive carbohydrate digestion in the presence of an active alpha-amylase.