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Pactacin is a novel digestive enzyme in teleosts.

ABSTRACT: Generally, animals extract nutrients from food by degradation using digestive enzymes. Trypsin and chymotrypsin, one of the major digestive enzymes in vertebrates, are pancreatic proenzymes secreted into the intestines. In this investigation, we report the identification of a digestive teleost enzyme, a pancreatic astacin that we termed pactacin. Pactacin, which belongs to the astacin metalloprotease family, emerged during the evolution of teleosts through gene duplication of astacin family enzymes containing six cysteine residues (C6astacin, or C6AST). In this study, we first cloned C6AST genes from pot-bellied seahorse (Hippocampus abdominalis) and analyzed their phylogenetic relationships using over 100 C6AST genes. Nearly all these genes belong to one of three clades: pactacin, nephrosin, and patristacin. Genes of the pactacin clade were further divided into three subclades. To compare the localization and functions of the three pactacin subclades, we studied pactacin enzymes in pot-bellied seahorse and medaka (Oryzias latipes). In situ hybridization revealed that genes of all three subclades were commonly expressed in the pancreas. Western blot analysis indicated storage of pactacin pro-enzyme form in the pancreas, and conversion to the active forms in the intestine. Finally, we partially purified the pactacin from digestive fluid, and found that pactacin is novel digestive enzyme that is specific in teleosts.

SUBMITTER: Kawaguchi M

PROVIDER: S-EPMC8012361 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Pactacin is a novel digestive enzyme in teleosts.

Kawaguchi Mari M Okazawa Yohei Y Imafuku Aiko A Nakano Yuko Y Shimizu Risa R Ishizuka Reiji R Jiang Tianlong T Nagasawa Tatsuki T Hiroi Junya J Yasumasu Shigeki S

Scientific reports 20210331 1

Generally, animals extract nutrients from food by degradation using digestive enzymes. Trypsin and chymotrypsin, one of the major digestive enzymes in vertebrates, are pancreatic proenzymes secreted into the intestines. In this investigation, we report the identification of a digestive teleost enzyme, a pancreatic astacin that we termed pactacin. Pactacin, which belongs to the astacin metalloprotease family, emerged during the evolution of teleosts through gene duplication of astacin family enzy ...[more]

PMID: 33790319

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Project description:Adaptation to a hypertonic marine environment is one of the major topics in animal physiology research. Marine teleosts lose water osmotically from the gills and compensate for this loss by drinking surrounding seawater and absorbing water from the intestine. This situation is in contrast to that in mammals, which experience a net osmotic loss of water after drinking seawater. Water absorption in fishes is made possible by (1) removal of monovalent ions (desalinization) by the esophagus, (2) removal of divalent ions as carbonate (Mg/CaCO3) precipitates promoted by HCO3- secretion, and (3) facilitation of NaCl and water absorption from diluted seawater by the intestine using a suite of unique transporters. As a result, 70-85% of ingested seawater is absorbed during its passage through the digestive tract. Thus, the digestive tract is an essential organ for marine teleost survival in the hypertonic seawater environment. The eel is a species that has been frequently used for osmoregulation research in laboratories worldwide. The eel possesses many advantages as an experimental animal for osmoregulation studies, one of which is its outstanding euryhalinity, which enables researchers to examine changes in the structure and function of the digestive tract after direct transfer from freshwater to seawater. In recent years, the molecular mechanisms of ion and water transport across epithelial cells (the transcellular route) and through tight junctions (the paracellular route) have been elucidated for the esophagus and intestine. Thanks to the rapid progress in analytical methods for genome databases on teleosts, including the eel, the molecular identities of transporters, channels, pumps and junctional proteins have been clarified at the isoform level. As 10 y have passed since the previous reviews on this subject, it seems relevant and timely to summarize recent progress in research on the molecular mechanisms of water and ion transport in the digestive tract in eels and to compare the mechanisms with those of other teleosts and mammals from comparative and evolutionary viewpoints. We also propose future directions for this research field to achieve integrative understanding of the role of the digestive tract in adaptation to seawater with regard to pathways/mechanisms including the paracellular route, divalent ion absorption, metabolon formation and cellular trafficking of transporters. Notably, some of these have already attracted practical attention in laboratories.

Project description:This study investigated the effects of multi-enzyme supplementation on various aspects of laying hens, including laying performance, egg quality, intestinal health and digestive function. In total, 384 Jingfen No.6 laying hens at 65-week-age were randomly assigned to four distinct dietary treatments: a basal diet (CON), CON supplemented with 150 g/t multi-enzyme (T1), CON with 300 g/t multi-enzyme (T2), and 600 g/t multi-enzyme (T3). A significant linear (P = 0.044) and quadratic (P = 0.014) increase was observed in the laying rate, while the feed/egg ratio exhibited a linear (P = 0.001) and quadratic (P < 0.001) decrease with increasing multi-enzyme supplementation. Additionally, linear (P < 0.05) and quadratic (P < 0.05) increases were observed in yolk rate and haugh unit with increasing levels of multi-enzyme supplementation. The trypsin activity in the duodenum and crude protein digestibility showed linear (P < 0.05) and quadratic (P < 0.05) increase with the addition of multi-enzyme. Furthermore, lipase and amylase activities in the duodenum increased quadratically (P = 0.041) and linearly (P = 0.040), respectively. Both jejunal and ileal digesta viscosities showed linear (P < 0.05) and quadratic (P < 0.05) decrease with the increasing addition of multi-enzyme. Moreover, multi-enzyme supplementation significantly increased (P < 0.05) the number of goblet cells in the intestinal of the treatment groups. The mRNA expression of Occludin-1, mucin 2 (MUC-2), large neutral amino acids transporter small subunit 1 (LAT-1) in the jejunum were significantly increased (P < 0.05) in the treatment groups (T1, T2 and T3) compared to the CON group. Additionally, the mRNA expression of solute carrier family 6-member 19 (B0AT-1) and large neutral amino acids transporter small subunit 4 (LAT-4) were significantly evaluated (P < 0.05) in the T2 and T3 groups, respectively. In conclusion, multi-enzyme supplementation enhanced digestive enzyme activities and intestinal barrier function, reduced intestinal digesta viscosity, and regulated mRNA expression of intestinal amino acid and lipid transporter genes, thereby improving crude protein digestibility and positively affecting laying performance and egg quality in hens.

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Pactacin is a novel digestive enzyme in teleosts.

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Pactacin is a novel digestive enzyme in teleosts.

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