Literature summary for 3.2.1.B44 extracted from

Xu, Y.; Feng, X.; Jia, J.; Chen, X.; Jiang, T.; Rasool, A.; Lv, B.; Qu, L.; Li, C.
A novel beta-glucuronidase from Talaromyces pinophilus Li-93 precisely hydrolyzes glycyrrhizin into glycyrrhetinic acid 3-O-mono-bea-D-glucuronide (2018), Appl. Environ. Microbiol., 84, e00755-18 .

Search for more literature for 3.2.1.B44

Activating Compound

Activating Compound	Comment	Organism
dithiothreitol	10 mM, about 1.1fold activation	Talaromyces pinophilus
EDTA	10 mM, about 1.1fold activation	Talaromyces pinophilus
glutathione	10 mM, about 1.2fold activation	Talaromyces pinophilus

Application

Application	Comment	Organism
synthesis	the enzyme is potentially a powerful biocatalyst for environmentally friendly and cost-effective production of glycyrrhetinic acid 3-O-mono-beta-D-glucuronide	Talaromyces pinophilus

Cloned(Commentary)

Cloned (Comment)	Organism
heterologous expression in Pichia pastoris	Talaromyces pinophilus

Protein Variants

Protein Variants	Comment	Organism
C331A	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at less than 5% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
C331D	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 25% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
C331H	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 30% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
C331K	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 25% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
C331L	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 65% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
C331S	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 30% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
H365A	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 40% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
H365C	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 50% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
H365D	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 20% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
H365K	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 60% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
H365L	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 70% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
H365S	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 50% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y330A	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 5% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y330C	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 40% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y330D	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 45% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y330F	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 40% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y330N	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 55% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y330R	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 55% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y372A	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at less than 5% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y372C	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 60% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y372D	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 20% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y372K	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 20% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y372L	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 70% of the activity compared to the wild-type enzyme	Talaromyces pinophilus
Y372S	the mutant enzyme catalyzes the hydrolysis of glycyrrhizin at about 55% of the activity compared to the wild-type enzyme	Talaromyces pinophilus

Inhibitors

Inhibitors	Comment	Organism
Al3+	10 mM, more than 90% loss of activity	Talaromyces pinophilus
Ca2+	10 mM, about 35% loss of activity	Talaromyces pinophilus
Cu2+	10 mM, more than 90% loss of activity	Talaromyces pinophilus
Fe2+	10 mM, more than 90% loss of activity	Talaromyces pinophilus
Mn2+	10 mM, about 40% loss of activity	Talaromyces pinophilus
Ni2+	10 mM, about 35% loss of activity	Talaromyces pinophilus
SDS	5 mM, about 70% loss of activity	Talaromyces pinophilus
Triton X-100	5 mM, about 40% loss of activity	Talaromyces pinophilus
Zn2+	10 mM, about 60% loss of activity	Talaromyces pinophilus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.54	-	glycyrrhizin	pH 4.5, 45°C	Talaromyces pinophilus
0.76	-	quercetin 3-O-beta-D-glucuronide	pH 4.5, 45°C	Talaromyces pinophilus
6.88	-	Baicalin	pH 4.5, 45°C	Talaromyces pinophilus
25.56	-	4-nitrophenyl beta-D-glucuronopyranoside	pH 4.5, 45°C	Talaromyces pinophilus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	-	Talaromyces pinophilus	-	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
47200	-	calculated from sequence	Talaromyces pinophilus
85000	-	SDS-PAGE	Talaromyces pinophilus

Organism

Organism	UniProt	Comment	Textmining
Talaromyces pinophilus	-	-	-
Talaromyces pinophilus Li-93	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	highly N-glycosylated protein	Talaromyces pinophilus

Purification (Commentary)

Purification (Comment)	Organism
-	Talaromyces pinophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
11.97	-	pH 4.5, 45°C	Talaromyces pinophilus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-nitrophenyl beta-D-glucuronopyranoside + H2O	at 5.8% as compared to hydrolysis of glycyrrhizin	Talaromyces pinophilus	4-nitrophenol + D-glucuronate	-	?
4-nitrophenyl beta-D-glucuronopyranoside + H2O	at 5.8% as compared to hydrolysis of glycyrrhizin	Talaromyces pinophilus Li-93	4-nitrophenol + D-glucuronate	-	?
baicalin + H2O	at 13.2% as compared to hydrolysis of glycyrrhizin	Talaromyces pinophilus	5,6,7-trihydroxyflavone + D-glucuronate	-	?
baicalin + H2O	at 13.2% as compared to hydrolysis of glycyrrhizin	Talaromyces pinophilus Li-93	5,6,7-trihydroxyflavone + D-glucuronate	-	?
glycyrrhizin + H2O	the enzyme recognizes and hydrolyzed the distal glucuronic bond of glycyrrhizin but can not cleave the glucuronic bond in glycyrrhetinic acid 3-O-mono-beta-D-glucuronide	Talaromyces pinophilus	glycyrrhetinate 3-O-beta-D-glucuronide + D-glucuronate	-	?
glycyrrhizin + H2O	the enzyme recognizes and hydrolyzed the distal glucuronic bond of glycyrrhizin but can not cleave the glucuronic bond in glycyrrhetinic acid 3-O-mono-beta-D-glucuronide	Talaromyces pinophilus Li-93	glycyrrhetinate 3-O-beta-D-glucuronide + D-glucuronate	-	?
quercetin 3-O-beta-D-glucuronide + H2O	at 51.2% as compared to hydrolysis of glycyrrhizin	Talaromyces pinophilus	quercetin + D-glucuronate	-	?
quercetin 3-O-beta-D-glucuronide + H2O	at 51.2% as compared to hydrolysis of glycyrrhizin	Talaromyces pinophilus Li-93	quercetin + D-glucuronate	-	?

Subunits

Subunits	Comment	Organism
monomer	1 * 85000, SDS-PAGE, N-glycosylated protein	Talaromyces pinophilus

Synonyms

Synonyms	Comment	Organism
glycyrrhizin glucuronohydrolase (glycyrrhetinic acid 3-O-mono-beta-D-glucuronide-forming)	-	Talaromyces pinophilus
TCE0_044r16781	-	Talaromyces pinophilus
TpGUS79A	-	Talaromyces pinophilus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	-	Talaromyces pinophilus

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
35	60	35°C: about 50% of maximal activity, 60°C: about 40% of maximal activity	Talaromyces pinophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
40	-	30 min, stable	Talaromyces pinophilus
60	-	30 min, the enzyme loses most of its activity	Talaromyces pinophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.26	-	4-nitrophenyl beta-D-glucuronopyranoside	pH 4.5, 45°C	Talaromyces pinophilus
2.9	-	Baicalin	pH 4.5, 45°C	Talaromyces pinophilus
2.9	-	quercetin 3-O-beta-D-glucuronide	pH 4.5, 45°C	Talaromyces pinophilus
6	-	glycyrrhizin	pH 4.5, 45°C	Talaromyces pinophilus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4.5	-	-	Talaromyces pinophilus

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	5.5	pH 4.0: about 30% of maximal activity, pH 5.5: about 30% of maximal activity	Talaromyces pinophilus

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
2	7.5	the enzyme is stable in the pH range of 2.0 to 7.5 at 4°C for 12 h	Talaromyces pinophilus

Expression

Organism	Comment	Expression
Talaromyces pinophilus	induced by glycyrrhizin	up

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.01	-	4-nitrophenyl beta-D-glucuronopyranoside	pH 4.5, 45°C	Talaromyces pinophilus
0.29	-	Baicalin	pH 4.5, 45°C	Talaromyces pinophilus
3.83	-	quercetin 3-O-beta-D-glucuronide	pH 4.5, 45°C	Talaromyces pinophilus
11.14	-	glycyrrhizin	pH 4.5, 45°C	Talaromyces pinophilus