cDNA sequences of GHF9 endo-β-1,4-glucanases in terrestrial Crustacea
| Autor: | Michael C. Gray, Stuart M. Linton, Benjamin J. Allardyce |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Signal peptide Coenobita brevimanus Brachyura 010603 evolutionary biology 01 natural sciences Isozyme Arthropod Proteins Evolution Molecular Open Reading Frames 03 medical and health sciences Cellulase Catalytic Domain Gene Duplication Complementary DNA Genetics Animals Cloning Molecular Gene Phylogeny biology Gene Expression Profiling Sequence Analysis DNA General Medicine Glucanase biology.organism_classification Open reading frame 030104 developmental biology Biochemistry Mictyris platycheles |
| Zdroj: | Gene. 642:408-422 |
| ISSN: | 0378-1119 |
| Popis: | This study aimed to sequence and identify a glycosyl hydrolase family 9 (GHF9) endo-β-1,4-glucanase expressed in the midgut gland of the herbivorous gecarcinid land crab, Gecarcoidea natalis. Hence this would explain the gene responsible for the production of previously purified and characterised endo-β-1,4-glucanases. Three different transcripts, two complete and one partial were sequenced from cDNA and an open reading frame of 1383bp was produced. Translated, this would produce a putative protein of 460 amino acid residues, including a 16 amino acid residue signal peptide. The mature protein (without signal peptide) is predicted to have a molecular mass of 47.6-47.7kDa; this closely matches the molecular mass (47.4kDa) of one of the three endo-β-1,4-glucanase/lichenase enzymes purified previously from G. natalis. It is therefore proposed that the gene described here encodes one of the previously characterised enzymes. The presence of multiple transcripts suggests gene duplication. To confirm that the gene is widely expressed within the Crustacea, cDNA encoding a GHF9 endo-β-1,4-glucanase was also sequenced in diverse crustaceans, the deposit feeding soldier crab, Mictyris platycheles and the terrestrial hermit crabs, Coenobita purlatus and C. brevimanus. An open reading frame of 1356bp was sequence from M. platycheles, while an incomplete open reading frames of 1384 and 1523bp were respectively sequenced from Coenobita brevimanus and C. perlatus. The midgut gland of M. platycheles contained activity (0.704±0.218μmol reducing sugars produced. min-1·mg-1 tissue wet weight) of a 26.3±0.3(5) endo-β-1,4-glucanase isozyme (determined from activity staining). These species, particularly M. platycheles does not consume and digest significant amounts of plant cellulose. This implies that the ancestral enzyme is not a cellulase, but rather it may be involved in hydrolysing cellulose like polysaccharides within other organisms such as algae. |
| Databáze: | OpenAIRE |
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