An eleven amino acid residue deletion expands the substrate specificity of acetyl xylan esterase II (AXE II) from Penicillium purpurogenum

Marcela Colombres, José A. Garate, Carlos F. Lagos, Raúl Araya-Secchi, Patricia Norambuena, Soledad Quiroz, Luis Larrondo, Tomas Pérez-Acle, Jaime Eyzaguirre

Resultado de la investigación: Article

6 Citas (Scopus)

Resumen

The soft-rot fungus Penicillium purpurogenum secretes to the culture medium a variety of enzymes related to xylan biodegradation, among them three acetyl xylan esterases (AXE I, II and III). AXE II has 207 amino acids; it belongs to family 5 of the carbohydrate esterases and its structure has been determined by X-ray crystallography at 0.9 Å resolution (PDB 1G66). The enzyme possesses the α/β hydrolase fold and the catalytic triad typical of serine esterases (Ser90, His187 and Asp175). AXE II can hydrolyze esters of a large variety of alcohols, but it is restricted to short chain fatty acids. An analysis of its three-dimensional structure shows that a loop that covers the active site may be responsible for this strict specificity. Cutinase, an enzyme that hydrolyzes esters of long chain fatty acids and shows a structure similar to AXE II, lacks this loop. In order to generate an AXE II with this broader specificity, the preparation of a mutant lacking residues involving this loop (Gly104 to Ala114) was proposed. A set of molecular simulation experiments based on a comparative model of the mutant enzyme predicted a stable structure. Using site-directed mutagenesis, the loop's residues have been eliminated from the AXE II cDNA. The mutant protein has been expressed in Aspergillus nidulans A722 and Pichia pastoris, and it is active towards a range of fatty acid esters of up to at least 14 carbons. The availability of an esterase with broader specificity may have biotechnological applications for the synthesis of sugar esters.

Idioma originalEnglish
Páginas (desde-hasta)19-28
Número de páginas10
PublicaciónJournal of Computer-Aided Molecular Design
Volumen22
N.º1
DOI
EstadoPublished - 1 ene 2008

Huella dactilar

acetylxylan esterase
deletion
Penicillium
Substrate Specificity
amino acids
Amino acids
enzymes
esters
fatty acids
Amino Acids
Esters
Substrates
Esterases
Enzymes
Aspergillus
Fatty acids
mutagenesis
biodegradation
culture media
fungi

ASJC Scopus subject areas

  • Molecular Medicine

Citar esto

Colombres, Marcela ; Garate, José A. ; Lagos, Carlos F. ; Araya-Secchi, Raúl ; Norambuena, Patricia ; Quiroz, Soledad ; Larrondo, Luis ; Pérez-Acle, Tomas ; Eyzaguirre, Jaime. / An eleven amino acid residue deletion expands the substrate specificity of acetyl xylan esterase II (AXE II) from Penicillium purpurogenum. En: Journal of Computer-Aided Molecular Design. 2008 ; Vol. 22, N.º 1. pp. 19-28.
@article{3d7f02c228d149a28debfcfffff85891,
title = "An eleven amino acid residue deletion expands the substrate specificity of acetyl xylan esterase II (AXE II) from Penicillium purpurogenum",
abstract = "The soft-rot fungus Penicillium purpurogenum secretes to the culture medium a variety of enzymes related to xylan biodegradation, among them three acetyl xylan esterases (AXE I, II and III). AXE II has 207 amino acids; it belongs to family 5 of the carbohydrate esterases and its structure has been determined by X-ray crystallography at 0.9 {\AA} resolution (PDB 1G66). The enzyme possesses the α/β hydrolase fold and the catalytic triad typical of serine esterases (Ser90, His187 and Asp175). AXE II can hydrolyze esters of a large variety of alcohols, but it is restricted to short chain fatty acids. An analysis of its three-dimensional structure shows that a loop that covers the active site may be responsible for this strict specificity. Cutinase, an enzyme that hydrolyzes esters of long chain fatty acids and shows a structure similar to AXE II, lacks this loop. In order to generate an AXE II with this broader specificity, the preparation of a mutant lacking residues involving this loop (Gly104 to Ala114) was proposed. A set of molecular simulation experiments based on a comparative model of the mutant enzyme predicted a stable structure. Using site-directed mutagenesis, the loop's residues have been eliminated from the AXE II cDNA. The mutant protein has been expressed in Aspergillus nidulans A722 and Pichia pastoris, and it is active towards a range of fatty acid esters of up to at least 14 carbons. The availability of an esterase with broader specificity may have biotechnological applications for the synthesis of sugar esters.",
keywords = "Acetyl xylan esterase, Comparative modeling, Cutinase, Mutagenesis, Penicillium purpurogenum",
author = "Marcela Colombres and Garate, {Jos{\'e} A.} and Lagos, {Carlos F.} and Ra{\'u}l Araya-Secchi and Patricia Norambuena and Soledad Quiroz and Luis Larrondo and Tomas P{\'e}rez-Acle and Jaime Eyzaguirre",
year = "2008",
month = "1",
day = "1",
doi = "10.1007/s10822-007-9149-4",
language = "English",
volume = "22",
pages = "19--28",
journal = "Journal of Computer-Aided Molecular Design",
issn = "0920-654X",
publisher = "Springer Netherlands",
number = "1",

}

An eleven amino acid residue deletion expands the substrate specificity of acetyl xylan esterase II (AXE II) from Penicillium purpurogenum. / Colombres, Marcela; Garate, José A.; Lagos, Carlos F.; Araya-Secchi, Raúl; Norambuena, Patricia; Quiroz, Soledad; Larrondo, Luis; Pérez-Acle, Tomas; Eyzaguirre, Jaime.

En: Journal of Computer-Aided Molecular Design, Vol. 22, N.º 1, 01.01.2008, p. 19-28.

Resultado de la investigación: Article

TY - JOUR

T1 - An eleven amino acid residue deletion expands the substrate specificity of acetyl xylan esterase II (AXE II) from Penicillium purpurogenum

AU - Colombres, Marcela

AU - Garate, José A.

AU - Lagos, Carlos F.

AU - Araya-Secchi, Raúl

AU - Norambuena, Patricia

AU - Quiroz, Soledad

AU - Larrondo, Luis

AU - Pérez-Acle, Tomas

AU - Eyzaguirre, Jaime

PY - 2008/1/1

Y1 - 2008/1/1

N2 - The soft-rot fungus Penicillium purpurogenum secretes to the culture medium a variety of enzymes related to xylan biodegradation, among them three acetyl xylan esterases (AXE I, II and III). AXE II has 207 amino acids; it belongs to family 5 of the carbohydrate esterases and its structure has been determined by X-ray crystallography at 0.9 Å resolution (PDB 1G66). The enzyme possesses the α/β hydrolase fold and the catalytic triad typical of serine esterases (Ser90, His187 and Asp175). AXE II can hydrolyze esters of a large variety of alcohols, but it is restricted to short chain fatty acids. An analysis of its three-dimensional structure shows that a loop that covers the active site may be responsible for this strict specificity. Cutinase, an enzyme that hydrolyzes esters of long chain fatty acids and shows a structure similar to AXE II, lacks this loop. In order to generate an AXE II with this broader specificity, the preparation of a mutant lacking residues involving this loop (Gly104 to Ala114) was proposed. A set of molecular simulation experiments based on a comparative model of the mutant enzyme predicted a stable structure. Using site-directed mutagenesis, the loop's residues have been eliminated from the AXE II cDNA. The mutant protein has been expressed in Aspergillus nidulans A722 and Pichia pastoris, and it is active towards a range of fatty acid esters of up to at least 14 carbons. The availability of an esterase with broader specificity may have biotechnological applications for the synthesis of sugar esters.

AB - The soft-rot fungus Penicillium purpurogenum secretes to the culture medium a variety of enzymes related to xylan biodegradation, among them three acetyl xylan esterases (AXE I, II and III). AXE II has 207 amino acids; it belongs to family 5 of the carbohydrate esterases and its structure has been determined by X-ray crystallography at 0.9 Å resolution (PDB 1G66). The enzyme possesses the α/β hydrolase fold and the catalytic triad typical of serine esterases (Ser90, His187 and Asp175). AXE II can hydrolyze esters of a large variety of alcohols, but it is restricted to short chain fatty acids. An analysis of its three-dimensional structure shows that a loop that covers the active site may be responsible for this strict specificity. Cutinase, an enzyme that hydrolyzes esters of long chain fatty acids and shows a structure similar to AXE II, lacks this loop. In order to generate an AXE II with this broader specificity, the preparation of a mutant lacking residues involving this loop (Gly104 to Ala114) was proposed. A set of molecular simulation experiments based on a comparative model of the mutant enzyme predicted a stable structure. Using site-directed mutagenesis, the loop's residues have been eliminated from the AXE II cDNA. The mutant protein has been expressed in Aspergillus nidulans A722 and Pichia pastoris, and it is active towards a range of fatty acid esters of up to at least 14 carbons. The availability of an esterase with broader specificity may have biotechnological applications for the synthesis of sugar esters.

KW - Acetyl xylan esterase

KW - Comparative modeling

KW - Cutinase

KW - Mutagenesis

KW - Penicillium purpurogenum

UR - http://www.scopus.com/inward/record.url?scp=38649094597&partnerID=8YFLogxK

U2 - 10.1007/s10822-007-9149-4

DO - 10.1007/s10822-007-9149-4

M3 - Article

C2 - 18060506

AN - SCOPUS:38649094597

VL - 22

SP - 19

EP - 28

JO - Journal of Computer-Aided Molecular Design

JF - Journal of Computer-Aided Molecular Design

SN - 0920-654X

IS - 1

ER -