Literature summary for 3.2.1.78 extracted from

dos Santos, C.; Paiva, J.; Meza, A.; Cota, J.; Alvarez, T.; Ruller, R.; Prade, R.; Squina, F.; Murakami, M.
Molecular insights into substrate specificity and thermal stability of a bacterial GH5-CBM27 endo-1,4-beta-D-mannanase (2012), J. Struct. Biol., 177, 469-476.

Cloned(Commentary)

Cloned (Comment)	Organism
gene Tpet_1542, recombinant expression of His-tagged full-length enzyme and isolated catalytic domain in Escerichia coli strain BL21(DE3)DELTASlyD	Thermotoga petrophila

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant isolated His-tagged catalytic domain in apoform and complexed with iodine, glucose, maltose, and maltose + gycerol, sitting drop vapor diffusion method, mixing of 0.0005 ml of 12 mg/ml protein in 25 mM Tris-HCl, pH 7.5, with 0.0005 ml of precipitant solution containing 0.1 M citrate, pH 5.5, 1 M ammonium phosphate, and 0.2 M sodium chloride, 20°C, soaking of crystals in ligand solutions, X-ray diffraction structure determination and analysis at 1.40-1,92 A resolution	Thermotoga petrophila

Crystallization (Comment)

Organism

purified recombinant isolated His-tagged catalytic domain in apoform and complexed with iodine, glucose, maltose, and maltose + gycerol, sitting drop vapor diffusion method, mixing of 0.0005 ml of 12 mg/ml protein in 25 mM Tris-HCl, pH 7.5, with 0.0005 ml of precipitant solution containing 0.1 M citrate, pH 5.5, 1 M ammonium phosphate, and 0.2 M sodium chloride, 20°C, soaking of crystals in ligand solutions, X-ray diffraction structure determination and analysis at 1.40-1,92 A resolution

Thermotoga petrophila

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Thermotoga petrophila	the enzyme catalyzes the hydrolysis of beta-1,4-mannoside linkages in various mannan-containing polysaccharides, such as glucomannans and galactomannans. Protein-ligand interactions from crystal structure analysis, circular dichroism spectroscopy, overview	?	-	?
additional information	Thermotoga petrophila RKU-1	the enzyme catalyzes the hydrolysis of beta-1,4-mannoside linkages in various mannan-containing polysaccharides, such as glucomannans and galactomannans. Protein-ligand interactions from crystal structure analysis, circular dichroism spectroscopy, overview	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermotoga petrophila	A5IMX7	gene Tpet_1542	-
Thermotoga petrophila RKU-1	A5IMX7	gene Tpet_1542	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged full-length enzyme and isolated catalytic domain from Escerichia coli strain BL21(DE3)DELTASlyD by nickel affinity chromatography, gel filtration, and ultrafiltration	Thermotoga petrophila

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the enzyme catalyzes the hydrolysis of beta-1,4-mannoside linkages in various mannan-containing polysaccharides, such as glucomannans and galactomannans. Protein-ligand interactions from crystal structure analysis, circular dichroism spectroscopy, overview	Thermotoga petrophila	?	-	?
additional information	the enzyme catalyzes the hydrolysis of beta-1,4-mannoside linkages in various mannan-containing polysaccharides, such as glucomannans and galactomannans. Protein-ligand interactions from crystal structure analysis, circular dichroism spectroscopy, overview	Thermotoga petrophila RKU-1	?	-	?

Subunits

Subunits	Comment	Organism
More	the two-domain enzyme encompasses a GH5 catalytic domain with a C-terminal CBM27 accessory domain, structure analysis and comparisons, overview	Thermotoga petrophila

Synonyms

Synonyms	Comment	Organism
Beta-mannanase	-	Thermotoga petrophila
endo-1,4-beta-D-mannanase	-	Thermotoga petrophila
GH5-CBM27	-	Thermotoga petrophila
TpMan	-	Thermotoga petrophila

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	the CBM27 accessory domain is critical for thermal stability, it reduces the melting temperature from 100°C to 88°C	Thermotoga petrophila

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycosyl hydrolase family 5, GH5	Thermotoga petrophila
additional information	the structure of the catalytic domain reveals a canonical (alpha/beta)8-barrel scaffold surrounded by loops and short helices that form the catalytic interface, subsites forming the active-site cleft with residues W134, E198, R200, E235, H283 and W284 are directly involved in glucose binding, structure analysis of full-length enzyme and catalytic domain, overview	Thermotoga petrophila