Literature summary for 3.2.1.21 extracted from

Toonkool, P.; Metheenukul, P.; Sujiwattanarat, P.; Paiboon, P.; Tongtubtim, N.; Ketudat-Cairns, M.; Ketudat-Cairns, J.; Svasti, J.
Expression and purification of dalcochinase, a beta-glucosidase from Dalbergia cochinchinensis Pierre, in yeast and bacterial hosts (2006), Protein Expr. Purif., 48, 195-204.

Cloned(Commentary)

Cloned (Comment)	Organism
enzyme with an N-terminal truncation following the propeptide and eight histidine residues enabling its purification by immobilized metal-ion affnity chromatography, functional, methanol-inducible enzyme expression in Pichia pastoris strain GS115 and Saccharomyces cerevisiae strains DY150 and DBY746, addition of 0.5% casamino acids to the culture medium stabilizes the pH of the culture and increaseds the protein yield by 3-5folds, insertion of a polyhistidine-tag either after the N-terminal alpha-factor signal sequence or at the C-terminus fails to assist in purification by immobilized metal-ion affnity chromatography due to post-translational processing at both termini, expression in Escherichia coli strain BL21(DE3) as insoluble and inactive protein, which can be made soluble by co-expression with bacterial chaperonin GroESL	Dalbergia cochinchinensis

Cloned (Comment)

Organism

enzyme with an N-terminal truncation following the propeptide and eight histidine residues enabling its purification by immobilized metal-ion affnity chromatography, functional, methanol-inducible enzyme expression in Pichia pastoris strain GS115 and Saccharomyces cerevisiae strains DY150 and DBY746, addition of 0.5% casamino acids to the culture medium stabilizes the pH of the culture and increaseds the protein yield by 3-5folds, insertion of a polyhistidine-tag either after the N-terminal alpha-factor signal sequence or at the C-terminus fails to assist in purification by immobilized metal-ion affnity chromatography due to post-translational processing at both termini, expression in Escherichia coli strain BL21(DE3) as insoluble and inactive protein, which can be made soluble by co-expression with bacterial chaperonin GroESL

Dalbergia cochinchinensis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.54	-	4-nitrophenyl-beta-D-fucoside	pH 5.0, 30°C, native enzyme	Dalbergia cochinchinensis
0.57	-	4-nitrophenyl-beta-D-fucoside	pH 5.0, 30°C, recombinant enzyme	Dalbergia cochinchinensis
1.48	-	dalcochinin-8'-O-beta-D-glucoside	pH 5.0, 30°C, recombinant enzyme	Dalbergia cochinchinensis
1.68	-	dalcochinin-8'-O-beta-D-glucoside	pH 5.0, 30°C, native enzyme	Dalbergia cochinchinensis
5.37	-	4-nitrophenyl-beta-D-glucopyranoside	pH 5.0, 30°C, native enzyme	Dalbergia cochinchinensis
5.78	-	4-nitrophenyl-beta-D-glucopyranoside	pH 5.0, 30°C, recombinant enzyme	Dalbergia cochinchinensis

Organism

Organism	UniProt	Comment	Textmining
Dalbergia cochinchinensis	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
additional information	recombinant dalcochinase is apparently composed of differently post-translationally modified forms with similar properties compared to the native enzyme	Dalbergia cochinchinensis

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme 23.3fold from yeast by immobilized metal-ion affnity chromatography, following hydrophobic interaction chromatography, insertion of a polyhistidine-tag either after the N-terminal alpha-factor signal sequence or at the C-terminus fails to assist in purification by immobilized metal-ion affnity chromatography due to post-translational processing at both termini	Dalbergia cochinchinensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
13.8	-	purified recombinant, substrate 4-nitrophenyl-beta-D-glucopyranoside	Dalbergia cochinchinensis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-methylumbelliferyl-beta-D-glucopyranoside + H2O	-	Dalbergia cochinchinensis	4-methylumbelliferol + beta-D-glucose	-	?
4-nitrophenyl beta-D-fucoside + H2O	-	Dalbergia cochinchinensis	4-nitrophenol + beta-D-fucose	-	?
4-nitrophenyl beta-D-glucopyranoside + H2O	-	Dalbergia cochinchinensis	4-nitrophenol + beta-D-glucose	-	?
dalcochinin-8'-O-beta-D-glucoside + H2O	-	Dalbergia cochinchinensis	dalcochinin + beta-D-glucose	-	?

Subunits

Subunits	Comment	Organism
?	x * 63000-64000, native enzyme, SDS-PAGE, x * 66000-69000, recombinant dalcochinase with N-terminal truncation and polyhistidine tag, SDS-PAGE	Dalbergia cochinchinensis

Synonyms

Synonyms	Comment	Organism
dalcochinase	-	Dalbergia cochinchinensis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	60	-	Dalbergia cochinchinensis

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
20	70	-	Dalbergia cochinchinensis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
60	-	purified recombinant and native enzymes, stable up to for 50 min	Dalbergia cochinchinensis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5	6	-	Dalbergia cochinchinensis