Biochemical characterization, cloning and molecular modeling of a digestive lipase from red seabream (Pagrus major): Structural explanation of the interaction deficiency with colipase and lipidic interface

Abstract

Biochemical characterization, cloning and molecular modeling of a digestive lipase from red seabream (Pagrus major): Structural explanation of the interaction deficiency with colipase and lipidic interfaceRed seabream digestive lipase (RsDL) was purified from fresh pyloric caeca. Pure RsDL has an apparent molecular mass of 50 kDa. The RsDL is more active on shortchain triacylglycerols (TC 4 ), and enzymatic activity decreases when medium (TC 8 ) or long-chain (olive oil) triacylglycerols were used as substrates. The specific activities of RsDL are very weak as compared to those obtained with classical pancreatic lipases. No colipase was detected in the red seabream pyloric caeca. Furthermore, the RsDL was not activated by a mammal colipase. Similar results were reported for annular seabream lipase. In order to explain structurally the discrepancies between sparidae and mammal lipases, genes encoding mature RsDL and five other lipases from sparidae fish species were cloned and sequenced. Phylogenetic studies indicated the closest homology of sparidae lipases to bird pancreatic ones. Structural models were built for annular seabream and RsDL under their closed and open forms using mammal pancreatic lipases as templates. Several differences were noticed when analyzing the amino acids corresponding to those involved in HPL binding to colipase. This is likely to prevent interaction between the fish lipase and the mammalian colipase and may explain the fact that mammalian colipase is not effective in activating sparidae lipases. In addition, several hydrophobic residues, playing a key role in anchoring pancreatic lipase onto the lipid interface, are replaced by polar residues in fish lipases. This might explain the reason why the latter enzymes display weak activity levels when compared to mammalian pancreatic lipases.Abbreviations: AsDL, annular seabream digestive lipase; DL, digestive lipase; DPLRP1, dog pancreatic lipase related protein 1; GhDL, gilthead seabream digestive lipase; HPL, human pancreatic lipase; HPLRP1, human pancreatic lipase related protein 1; HPLRP2, human pancreatic lipase related protein 2; PCR, polymerase chain reaction; PL, pancreatic lipase; PPL, porcine pancreatic lipase; PPLRP2, porcine pancreatic lipase related protein 2; RP, related pancreatic lipase; RsDL, red seabream digestive lipase; RT, reverse transcription; SpDL, Saupe digestive lipase; SsDL, striped seabream digestive lipase; TPL, turkey pancreatic lipase; WsDL, white seabream digestive lipase

Comparative Study of the Molecular Characterization, Evolution, and Structure Modeling of Digestive Lipase Genes Reveals the Different Evolutionary Selection Between Mammals and Fishes

“…Similarly, the BSAL also did not occur in gene duplication in mammals. In addition, the amino acid of the lid domain in pancreatic lipase was conserved in these species, it also means that the function of pancreatic lipase might be fully-functioning ( Ollis et al, 1992 ; Verger 1997 ; Smichi et al, 2017 ).…”

“…Therefore, the copy number of lipase genes exhibited different models between mammals and fishes. Meanwhile, the amino acid sequence analysis showed that the Ile and Leu were replaced by Ser, Lys, or Arg in the lid domain of pancreatic lipase in most fishes species, it also might influence the lipolytic ability of pancreatic lipase in fish (Smichi et al, 2017). However, the bsal gene occurred the gene duplication and obtained two to five copies in the fish species in which the Ile and Leu were absent, the phylogenetic analysis also showed that these fish species were grouped together (Figure 3).…”

“…Although, most fishes had a plrp gene (a copy of pl ), according to some reports ( Sæle et al, 2010 ; Rønnestad et al, 2013 ) and annotation in NCBI, these plrp genes might produce inactive pancreatic lipases in teleosts, these pancreatic lipases might not possess the lipolytic function due to mutations in the domain ( Sæle et al, 2010 ; Sæle et al, 2018 ). Moreover, pancreatic lipase is a colipase-dependent lipase, colipase is required for lipid binding and allows the pancreatic lipase to carry out the function of hydrolyzing lipids ( Smichi et al, 2017 ; Anderson et al, 2018 ; González-Félix ML. et al, 2018 ; Achouri et al, 2020 ).…”

“…et al, 2018 ; Achouri et al, 2020 ). The absence of the colipase gene ( clps ) also can be found in zebrafish ( Danio rerio ), medaka ( Oryzias latipe ), Japanese pufferfish ( Takifugu rubripes ), and three-spined stickleback ( Gasterosteus aculeatus ) ( Sæle et al, 2010 ; Smichi et al, 2017 ; Anderson et al, 2018 ; Sæle et al, 2018 ).…”

Dissecting the Interaction Deficiency of a Cartilaginous Fish Digestive Lipase with Pancreatic Colipase: Biochemical and Structural Insights

“…The characterized enzyme shows distinct properties to TPL [8] and lipases from other marine species [19,21]. From the same family, a lipase from the red seabream has been isolated and purified and its functional properties have been performed to explain the differences with pancreatic lipases [25]. Yet, little information is available regarding structurefunction relationships of cartilaginous fish (chondrichthyans) lipases.…”