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Literature summary for 3.1.1.20 extracted from

  • Boeer, E.; Bode, R.; Mock, H.P.; Piontek, M.; Kunze, G.
    Atan1p-an extracellular tannase from the dimorphic yeast Arxula adeninivorans: molecular cloning of the ATAN1 gene and characterization of the recombinant enzyme (2009), Yeast, 26, 323-337.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant ATAN1 gene overexpressed in the auxotrophic mutant strain Arxula adeninivorans G1212 [DELTAatrp1] under the control of the strong, constitutive TEF1 promoter. Expression module assembled in the plasmid pGH101-TEF1-ATAN1 and used to construct YIEC101-TEF1-ATAN1 for transformation of Arxula host cells Blastobotrys adeninivorans

Inhibitors

Inhibitors Comment Organism Structure
Cd2+ 45% inhibition, at 30°C in sodium citrate buffer, pH 6.0 Blastobotrys adeninivorans
Cu2+ 51% inhibition, at 30°C in sodium citrate buffer, pH 6.0 Blastobotrys adeninivorans
Fe3+ 35% inhibition, at 30°C in sodium citrate buffer, pH 6.0 Blastobotrys adeninivorans
gallic acid competitive inhibition using gallic acid as substrate Blastobotrys adeninivorans
additional information EDTA, Mg2+, Mn2+, Ca2+, Zn2+ and EDTA have no or only weak effects Blastobotrys adeninivorans
phenyl methyl sulphonyl fluoride 95% inhibition, at 30°C in sodium citrate buffer, pH 6.0 Blastobotrys adeninivorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
Tannic acid endogenous tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans
0.17
-
Tannic acid recombinant tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans
3.5
-
methyl gallate recombinant tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans
4.4
-
methyl gallate endogenous tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
4 * 80000, denatured endogenous and recombinant tannases, SDS-PAGE. 4 * 55000, deglycosylated endogenous and recombinant tannases, SDS-PAGE Blastobotrys adeninivorans
61000
-
mature enzyme, calculated from the ORF Blastobotrys adeninivorans
80000
-
4 * 80000, denatured endogenous and recombinant tannases, SDS-PAGE. 4 * 55000, deglycosylated endogenous and recombinant tannases, SDS-PAGE Blastobotrys adeninivorans
320000
-
native endogenous and recombinant tannases, gel filtration Blastobotrys adeninivorans

Organism

Organism UniProt Comment Textmining
Blastobotrys adeninivorans B7VFD0
-
-
Blastobotrys adeninivorans LS3 B7VFD0
-
-

Purification (Commentary)

Purification (Comment) Organism
endogenous and recombinant tannases purified by ion-exchange chromatography and gel filtration Blastobotrys adeninivorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
catechin gallate + H2O best substrate Blastobotrys adeninivorans gallic acid + catechin
-
?
catechin gallate + H2O best substrate Blastobotrys adeninivorans LS3 gallic acid + catechin
-
?
epicatechin gallate + H2O
-
Blastobotrys adeninivorans gallic acid + epicatechin
-
?
epicatechin gallate + H2O
-
Blastobotrys adeninivorans LS3 gallic acid + epicatechin
-
?
epigallocatechin gallate + H2O
-
Blastobotrys adeninivorans gallic acid + epigallocatechin
-
?
epigallocatechin gallate + H2O
-
Blastobotrys adeninivorans LS3 gallic acid + epigallocatechin
-
?
gallic acid ethyl ester + H2O
-
Blastobotrys adeninivorans gallic acid + ethanol
-
?
gallic acid propyl ester + H2O
-
Blastobotrys adeninivorans gallic acid + propanol
-
?
gallocatechin gallate + H2O
-
Blastobotrys adeninivorans gallic acid + gallocatechin
-
?
gallocatechin gallate + H2O
-
Blastobotrys adeninivorans LS3 gallic acid + gallocatechin
-
?
methyl gallate + H2O
-
Blastobotrys adeninivorans gallate + methanol
-
?
methyl gallate + H2O
-
Blastobotrys adeninivorans LS3 gallate + methanol
-
?
tannic acid + H2O
-
Blastobotrys adeninivorans gallic acid + D-glucose
-
?

Subunits

Subunits Comment Organism
homotetramer 4 * 80000, denatured endogenous and recombinant tannases, SDS-PAGE. 4 * 55000, deglycosylated endogenous and recombinant tannases, SDS-PAGE Blastobotrys adeninivorans

Synonyms

Synonyms Comment Organism
ATAN1
-
Blastobotrys adeninivorans
tannase
-
Blastobotrys adeninivorans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35 40 endogenous and recombinant tannases Blastobotrys adeninivorans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 50 endogenous and recombinant tannases Blastobotrys adeninivorans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.7 6 endogenous and recombinant tannases Blastobotrys adeninivorans

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 7 endogenous and recombinant tannases Blastobotrys adeninivorans

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.45
-
gallic acid endogenous tannase Blastobotrys adeninivorans

Expression

Organism Comment Expression
Blastobotrys adeninivorans in the presence of tannic acid, ATAN1 transcript declines after 12 h of incubation, by 24 h titres of the ATAN1 transcript drop below the detection limit down
Blastobotrys adeninivorans ATAN1 is expressed only in the presence of tannic acid or gallic acid. Transcripts are first detected 2 h after the shift from glucose to tannic acid medium, levels then increase, reaching a maximum after 4-12 h. ATAN1 transcript is detectable within 30 min in the presence of gallic acid up

General Information

General Information Comment Organism
physiological function enzyme titre with the recombinant strain (390 U/l) is approximately 10times higher than that in the control strain without the TEF1 promoter-ATAN1 gene expression cassette. The Atan1 protein contains at positions 210-214 the canonical Gly-X-Ser-X-Gly motif found in the serine hydrolases as the catalytic triad for nucleophilic serine Blastobotrys adeninivorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.064
-
Tannic acid recombinant tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans
0.078
-
Tannic acid endogenous tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans
0.112
-
methyl gallate recombinant tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans
0.127
-
methyl gallate endogenous tannase, at 30°C, pH 6.0 Blastobotrys adeninivorans