Cloned (Comment) | Organism |
---|---|
gene man5, DNA and amino acid sequence determination and analysis | Neurospora sitophila |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion method, mixing of 6.5 mg/ml protein in 20 mM Tris-HCl, pH 7.6, with reservoir solution containing 0.1 M magnesium chloride, 0.1 M sodium acetate pH 4.5, 23% w/v PEG 3350, 20°C, method optimization, X-ray diffraction structure determination and analysis at 1.40 A resolution, molecular replacement | Neurospora sitophila |
Protein Variants | Comment | Organism |
---|---|---|
additional information | mutagenesis of Trp271 at the +1 subsite plays a crucial role in the enhanced transglycosylation activity | Neurospora sitophila |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Neurospora sitophila | the native wild-type enzyme shows bifunctional activity as both an endo-glucanase and an endo-mannanase, substrate specificity, overview | ? | - |
? | |
additional information | Neurospora sitophila DSM 16514 | the native wild-type enzyme shows bifunctional activity as both an endo-glucanase and an endo-mannanase, substrate specificity, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Neurospora sitophila | I4IY26 | i.e. Neurospora sitophila, gene man5 | - |
Neurospora sitophila DSM 16514 | I4IY26 | i.e. Neurospora sitophila, gene man5 | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | the enzyme shows two different N-glycosylation sites. The first site Asn63 contains two linked N-acetylglucosamine residues. This glycosylation may have a structural role in stabilizing structural elements, as there are hydrogen bonds between the glucosamine residues and Ile67 and Leu71, which both belong to a very long loop (residues 64-84), and Val136 located on helix 4. The other site Asn319 is not fully occupied, suggesting disorder or perhaps different populations of glycosylation | Neurospora sitophila |
Purification (Comment) | Organism |
---|---|
native enzyme by ultrafiltration and anion exchange chromatography | Neurospora sitophila |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | solid state fermentation on medium containing microcrystalline cellulose | Neurospora sitophila | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl beta-D-mannopyranoside + H2O | - |
Neurospora sitophila | 4-nitrophenol + beta-D-mannopyranose | - |
? | |
4-nitrophenyl beta-D-mannopyranoside + H2O | - |
Neurospora sitophila DSM 16514 | 4-nitrophenol + beta-D-mannopyranose | - |
? | |
azo-carob galactomannan + H2O | - |
Neurospora sitophila | ? | - |
? | |
azo-carob galactomannan + H2O | - |
Neurospora sitophila DSM 16514 | ? | - |
? | |
additional information | the native wild-type enzyme shows bifunctional activity as both an endo-glucanase and an endo-mannanase, substrate specificity, overview | Neurospora sitophila | ? | - |
? | |
additional information | the enzyme also show transglycosylation activity | Neurospora sitophila | ? | - |
? | |
additional information | the native wild-type enzyme shows bifunctional activity as both an endo-glucanase and an endo-mannanase, substrate specificity, overview | Neurospora sitophila DSM 16514 | ? | - |
? | |
additional information | the enzyme also show transglycosylation activity | Neurospora sitophila DSM 16514 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme adopts the (beta/alpha)8-barrel fold | Neurospora sitophila |
Synonyms | Comment | Organism |
---|---|---|
CsMan5 | - |
Neurospora sitophila |
endo-beta-D-1,4-mannanase | - |
Neurospora sitophila |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycosyl hydrolase family 5, GH5, structure comparisons, the overall fold of the enzyme is strongly conserved, overview. The enzyme displays the typical (beta/alpha)8-barrel fold and a unique structural arrangement of three surface loops that stretch over the active centre, promoting an altered topography of the binding cleft | Neurospora sitophila |
additional information | tertiary structure, active site and substrate binding site structures analysis, detailed overview. Two tryptophan residues that provide the hydrophobic stacking of the +1 subsite Trp125 and Trp271,is 9.5 A, making bulkier branched substrates difficult to accommodate | Neurospora sitophila |