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Literature summary for 3.1.1.1 extracted from

  • Rusnak, M.; Nieveler, J.M Schmid, R. D.; Petri, R.
    The putative lipase, AF1763, from Archaeoglobus fulgidus is a carboxylesterase with a very high pH optimum (2005), Biotechnol. Lett., 27, 743-748.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
-
Archaeoglobus fulgidus
expression in Escherichia coli Archaeoglobus fulgidus

Protein Variants

Protein Variants Comment Organism
D163N total loss of activity Archaeoglobus fulgidus
D163N inactive mutant enzyme Archaeoglobus fulgidus
D169N 45% loss of activity compared to wild-type enzyme Archaeoglobus fulgidus
D169N 50% of wild-type activity remains Archaeoglobus fulgidus
H210N total loss of activity Archaeoglobus fulgidus
H210N 95% loss of activity compared to wild-type enzyme Archaeoglobus fulgidus
S136C total loss of activity Archaeoglobus fulgidus
S136C inactive mutant enzyme Archaeoglobus fulgidus
Y244stop a truncated version of the enzyme covering the 244 N-terminal residues reveals measurable activity Archaeoglobus fulgidus
Y255stop 88% loss of activity compared to wild-type enzyme Archaeoglobus fulgidus

Inhibitors

Inhibitors Comment Organism Structure
additional information phenylmethansulfonfluoride has almost no influence on the activity up to 10 mM; PMSF has almost no influence on the activity of LipA up to 10 mM Archaeoglobus fulgidus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.876
-
4-nitrophenyl palmitate pH 7, 50°C Archaeoglobus fulgidus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52800
-
x * 52800, calculated from sequence Archaeoglobus fulgidus
52800
-
x * 52800, SDS-PAGE Archaeoglobus fulgidus

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus O28511
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O converted twice as fast as the corresponding palmitate Archaeoglobus fulgidus 4-nitrophenol + acetate
-
?
4-nitrophenyl palmitate + H2O
-
Archaeoglobus fulgidus 4-nitrophenol + palmitate
-
?
butyl acetate + H2O
-
Archaeoglobus fulgidus butanol + acetate
-
?
butyl acetate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus butanol + acetate
-
?
ethyl acetate + H2O
-
Archaeoglobus fulgidus ethanol + acetate
-
?
ethyl butyrate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus ethanol + butanoate
-
?
ethyl propionate + H2O
-
Archaeoglobus fulgidus ethanol + propionate
-
?
ethyl propionate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus ethanol + propionate
-
?
methyl acetate + H2O
-
Archaeoglobus fulgidus methanol + acetate
-
?
methyl acetate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus methanol + acetate
-
?
methyl butyrate + H2O preferred substrate. The enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus methanol + butanoate
-
?
methyl butyrate + H2O
-
Archaeoglobus fulgidus methanol + butyrate
-
?
methyl hexanoate + H2O
-
Archaeoglobus fulgidus methanol + hexanoate
-
?
methyl hexanoate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus methanol + hexanoate
-
?
methyl propionate + H2O
-
Archaeoglobus fulgidus methanol + propionate
-
?
methyl propionate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus methanol + propionate
-
?
additional information no activity detected with methyl myristate. Tributyrin is hydrolyzed by LipA at a lower rate than short and medium chain fatty acid esters and 4-nitrophenyl acetate is converted twice as fast as the corresponding palmitate Archaeoglobus fulgidus ?
-
?
additional information the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols. Methyl esters are hydrolyzed faster than the corresponding ethyl esters. Butyrate is the preferred chain length for the carboxylic acid. No activity with esters of hydroxy-substituted fatty acids such as 3-hydroxybutyrate Archaeoglobus fulgidus ?
-
?
phenyl acetate + H2O
-
Archaeoglobus fulgidus phenol + acetate
-
?
phenyl acetate + H2O the enzyme preferentially hydrolyzes esters of short-chain fatty acids (C2–C4) and especially those of medium chain (C5–C6) fatty acids with short chain alcohols Archaeoglobus fulgidus phenol + acetate
-
?
tributyrin + H2O hydrolyzed at a lower rate than short and medium chain fatty acid esters Archaeoglobus fulgidus ?
-
?

Subunits

Subunits Comment Organism
? x * 52800, calculated from sequence Archaeoglobus fulgidus
? x * 52800, SDS-PAGE Archaeoglobus fulgidus

Synonyms

Synonyms Comment Organism
AF1763
-
Archaeoglobus fulgidus
AF1763 locus name Archaeoglobus fulgidus
LipA
-
Archaeoglobus fulgidus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
-
Archaeoglobus fulgidus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 90 40°C: about 40% of maximal activity, 90°C: about 65% of maximal activity Archaeoglobus fulgidus
40 95 40°C: about 40% of maximal activity, 95°C: about 40% of maximal activity Archaeoglobus fulgidus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
about 60% loss of activity after 30 min, about 85% loss of activity after 90 min Archaeoglobus fulgidus
40
-
about 60% loss of activity after 30 min Archaeoglobus fulgidus
60
-
about 60% loss of activity after 30 min, about 85% loss of activity after 60 min Archaeoglobus fulgidus
60
-
about 60% loss of activity after 30 min Archaeoglobus fulgidus
80
-
30 min, complete loss of activity Archaeoglobus fulgidus
80
-
complete loss of activity after 30 min Archaeoglobus fulgidus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
47.5
-
4-nitrophenyl palmitate pH 7, 50°C Archaeoglobus fulgidus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
at 50°C Archaeoglobus fulgidus
10 11 at room temperature Archaeoglobus fulgidus

pH Range

pH Minimum pH Maximum Comment Organism
7 12 pH 7.0: about 40% of maximal activity, pH 12: about 30% of maximal activity, at room temperature Archaeoglobus fulgidus
8 12 pH 8: about 60% of maximal activity, pH 12: about 30% of maximal activity, at room temperature Archaeoglobus fulgidus
9 12 pH 9.0: about 70% of maximal activity, pH 12: about 60% of maximal activity, at 50°C Archaeoglobus fulgidus
9 12 pH 9: about 70% of maximal activity, pH 12: about 60% of maximal activity, at 50°C Archaeoglobus fulgidus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
54
-
4-nitrophenyl palmitate pH 7, 50°C Archaeoglobus fulgidus