Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged wild-type and selenomethionine-labeled enzyme in Escherichia coli | Rhizomucor miehei |
expression in Escherichia coli | Rhizomucor miehei |
Crystallization (Comment) | Organism |
---|---|
purified recombinant detagged wild-type and selenomethionine-labeled enzymes, sitting drop vapour diffusion method, mixing of 0.001 ml of 20 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 100 mM NaCl, with 0.001 ml of precipitant solution containing 200 mM Li2SO4, 30% w/v PEG 4000, 100 mM Tris-HCl, pH 8.5, or 160 mM Li2SO4, 24% w/v PEG 4000, 80 mM Tris-HCl, pH 8.5, 6% v/v 2-methyl-2,4-pentanediol, 2-4 weeks, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution | Rhizomucor miehei |
the overall structure of the Lam81A monomer consists of an N-terminal beta-sandwich domain, a C-terminal (alpha/alpha)6 domain and an additional domain between them. Residues Glu553 and Glu557 are proposed to serve as the proton donor and basic catalyst, respectively, in a single-displacement mechanism. In addition, Tyr386, Tyr482 and Ser554 possibly contribute to both the position or the ionization state of the basic catalyst Glu557 | Rhizomucor miehei |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhizomucor miehei | A0A023I7E1 | gene RmLam81A | - |
Rhizomucor miehei | U5HK45 | - |
- |
Rhizomucor miehei CAU432 | A0A023I7E1 | gene RmLam81A | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and selenomethionine-labeled enzymes from Escherichia coli by nickel affinity chromatography, ultrafiltration, tag cleavage by Bacillus subtilis proteases, and gel filtration | Rhizomucor miehei |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme monomer consists of an N-terminal beta-sandwich domain, a C-terminal (alpha/alpha)6 domain and an additional domain between them, three-dimensional structure of enzyme RmLam81A domain A, domain B and domain C, overview | Rhizomucor miehei |
Synonyms | Comment | Organism |
---|---|---|
endo-beta-1,3-glucanase | - |
Rhizomucor miehei |
GH family 81 beta-1,3-glucanase | - |
Rhizomucor miehei |
Lam81A | - |
Rhizomucor miehei |
laminarinase | - |
Rhizomucor miehei |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycosyl hydrolase family 81, GH81, but has a unique structure | Rhizomucor miehei |
additional information | the enzyme monomer consists of an N-terminal beta-sandwich domain, a C-terminal (alpha/alpha)6 domain and an additional domain between them. Glu553 and Glu557 are proposed to serve as the proton donor and basic catalyst, respectively, in a single-displacement mechanism. In addition, Tyr386, Tyr482 and Ser554 possibly contribute to both the position or the ionization state of the basic catalyst Glu557, catalytic cleft and the active site structure, overview | Rhizomucor miehei |