Isozymes from the herbivorous gecarcinid land crab, Gecarcoidea natalis that possess both lichenase and endo-β-1,4-glucanase activity

Linton, Stuart M.; Shirley, Alicia J.

doi:10.1016/j.cbpb.2011.05.007

Cited by 15 publications

(6 citation statements)

References 37 publications

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“…To do this, a β-1,3-glucanase sample, previously purified from the midgut gland of G. natalis, was run on a 12% polyacrylamide gel and stained with Coomassie blue as per the method of Allardyce and Linton(2008) and Linton and Shirley(2011). A protein band corresponding to the β-1,3-glucanase was excised from the gel and processed by in-gel tryptic digestion.…”

Section: Can the Putative Protein Be Translated Into A Digestive Enzyme?mentioning

confidence: 99%

A glycosyl hydrolase family 16 gene is responsible for the endogenous production of β-1,3-glucanases within decapod crustaceans

Linton

Cameron

Gray

et al. 2015

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To identify the gene responsible for the production of a β-1,3-glucanase (laminarinase) within crustacea, a glycosyl hydrolase family 16 (GHF16) gene was sequenced from the midgut glands of the gecarcinid land crab, Gecarcoidea natalis and the freshwater crayfish, Cherax destructor. An open reading frame of 1098 bp for G. natalis and 1095 bp for C. destructor was sequenced from cDNA. For G. natalis and C. destructor respectively, this encoded putative proteins of 365 and 364 amino acids with molecular masses of 41.4 and 41.5 kDa. mRNA for an identical GHF16 protein was also expressed in the haemolymph of C. destructor. These putative proteins contained binding and catalytic domains that are characteristic of a β-1,3-glucanase from glycosyl hydrolase family 16. The amino acid sequences of two short 8-9 amino acid residue peptides from a previously purified β-1,3-glucanase from G. natalis matched exactly that of the putative protein sequence. This plus the molecular masses of the putative proteins matching that of the purified proteins strongly suggests that the sequences obtained encode for a catalytically active β-1,3-glucanase. A glycosyl hydrolase family 16 cDNA was also partially sequenced from the midgut glands of other amphibious (Mictyris platycheles and Paragrapsus laevis) and terrestrial decapod species (Coenobita rugosus, Coenobita perlatus, Coenobita brevimanus and Birgus latro) to confirm that the gene is widely expressed within this group. There are three possible hypothesised functions and thus evolutionary routes for the β-1,3-glucanase: 1) a digestive enzyme which hydrolyses β-1,3-glucans, 2) an enzyme which cleaves β-1,3-glycosidic bonds within cell walls to release cell contents or 3) an immune protein which can hydrolyse the cell walls of potentially pathogenic micro-organisms.

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Section: Can the Putative Protein Be Translated Into A Digestive Enzyme?mentioning

confidence: 99%

A glycosyl hydrolase family 16 gene is responsible for the endogenous production of β-1,3-glucanases within decapod crustaceans

Linton

Cameron

Gray

et al. 2015

Gene

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“…This substrate is hydrolysed by enzymes that hydrolyse β-1,4-glycosidic bonds; this includes β-glucohydrolyses and endo-β-1,4-glucanases/lichenases (allardyce et al 2010). The large fluorescent band at 119-126 kDa is thought to be due to β-glucohydrolase, while the three smaller activity bands (32, 28 and 25 kDa) are thought to be endo-β-1,4-glucanase/lichenase isozymes that have been characterised previously (linton and Shirley 2011;allardyce and linton 2008). These enzymes when run on SDS-Page gels without heating or reducing the sample with β-mercaptoethanol, the conditions used to create the zymograms, run further than samples which were heated and reduced.…”

Section: Activity Staining For Enzymes That Hydrolyse β-14-glycosidimentioning

confidence: 99%

Digestive enzymes of two brachyuran and two anomuran land crabs from Christmas Island, Indian Ocean

et al. 2014

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higher endo-β-1,4-glucanase, lichenase and laminarinase activities compared to that of the other species. Thus, C. perlatus may be efficient at digestion of cellulose and hemicellulose within plant material. Zymography indicated that the majority of protease, lipase, phosphatase, amylase, endo-β-1,4-glucanase, β-glucohydrolase and N-acetyl-β-d-glucosaminidase isozymes were common to all species, and hence were inherited from a common aquatic ancestor. Differences were observed for the phosphatase, lipase and endo-β-1,4-glucanase isozymes. These differences are discussed in relation to phylogeny and possible evolution to cope with the adoption of a terrestrial diet.

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“…Investigation of sea hare digestion may therefore provide useful clues for establishment of an artificial process for saccharifying polysaccharides in sea lettuce. Although individual ß-1,4-endoglucanases have been purified from various marine animals, including crustaceans [13], [14], mollusks [15]–[18], and sea urchins [19], entire glucose production systems from polysaccharides in algae are yet to be clarified in higher animals, unlike in bacteria [20] and fungi [6]. Genome analysis and PCR cloning of glucanases using primers corresponding to conserved amino acid sequences show the existence of mRNA for a novel enzyme and elucidates the genetic evolution of glucanase [21]–[23].…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Enzymatic Analysis of the Cellulolytic System in Digestive Fluid of the Sea Hare Aplysia kurodai. Efficient Glucose Release from Sea Lettuce by Synergistic Action of 45 kDa Endoglucanase and 210 kDa ß-Glucosidase

et al. 2013

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Although many endo-ß-1,4-glucanases have been isolated in invertebrates, their cellulolytic systems are not fully understood. In particular, gastropod feeding on seaweed is considered an excellent model system for production of bioethanol and renewable bioenergy from third-generation feedstocks (microalgae and seaweeds). In this study, enzymes involved in the conversion of cellulose and other polysaccharides to glucose in digestive fluids of the sea hare (Aplysia kurodai) were screened and characterized to determine how the sea hare obtains glucose from sea lettuce (Ulva pertusa). Four endo-ß-1,4-glucanases (21K, 45K, 65K, and 95K cellulase) and 2 ß-glucosidases (110K and 210K) were purified to a homogeneous state, and the synergistic action of these enzymes during cellulose digestion was analyzed. All cellulases exhibited cellulase and lichenase activities and showed distinct cleavage specificities against cellooligosaccharides and filter paper. Filter paper was digested to cellobiose, cellotriose, and cellotetraose by 21K cellulase, whereas 45K and 65K enzymes hydrolyzed the filter paper to cellobiose and glucose. 210K ß-glucosidase showed unique substrate specificity against synthetic and natural substrates, and 4-methylumbelliferyl (4MU)-ß-glucoside, 4MU–ß-galactoside, cello-oligosaccharides, laminarin, and lichenan were suitable substrates. Furthermore, 210K ß-glucosidase possesses lactase activity. Although ß-glucosidase and cellulase are necessary for efficient hydrolysis of carboxymethylcellulose to glucose, laminarin is hydrolyzed to glucose only by 210K ß-glucosidase. Kinetic analysis of the inhibition of 210K ß-glucosidase by D-glucono-1,5-lactone suggested the presence of 2 active sites similar to those of mammalian lactase-phlorizin hydrolase. Saccharification of sea lettuce was considerably stimulated by the synergistic action of 45K cellulase and 210K ß-glucosidase. Our results indicate that 45K cellulase and 210K ß-glucosidase are the core components of the sea hare digestive system for efficient production of glucose from sea lettuce. These findings contribute important new insights into the development of biofuel processing biotechnologies from seaweed.

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Isozymes from the herbivorous gecarcinid land crab, Gecarcoidea natalis that possess both lichenase and endo-β-1,4-glucanase activity

Cited by 15 publications

References 37 publications

A glycosyl hydrolase family 16 gene is responsible for the endogenous production of β-1,3-glucanases within decapod crustaceans

A glycosyl hydrolase family 16 gene is responsible for the endogenous production of β-1,3-glucanases within decapod crustaceans

Digestive enzymes of two brachyuran and two anomuran land crabs from Christmas Island, Indian Ocean

Comprehensive Enzymatic Analysis of the Cellulolytic System in Digestive Fluid of the Sea Hare Aplysia kurodai. Efficient Glucose Release from Sea Lettuce by Synergistic Action of 45 kDa Endoglucanase and 210 kDa ß-Glucosidase

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