AXE2_GEOSE
ID AXE2_GEOSE Reviewed; 219 AA.
AC Q09LX1;
DT 14-OCT-2015, integrated into UniProtKB/Swiss-Prot.
DT 17-OCT-2006, sequence version 1.
DT 25-MAY-2022, entry version 51.
DE RecName: Full=Acetylxylan esterase {ECO:0000303|PubMed:21994937};
DE EC=3.1.1.72 {ECO:0000269|PubMed:21994937};
DE AltName: Full=Acetyl-xylooligosaccharide esterase {ECO:0000303|PubMed:24531461};
GN Name=axe2 {ECO:0000303|PubMed:21994937};
OS Geobacillus stearothermophilus (Bacillus stearothermophilus).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Geobacillus.
OX NCBI_TaxID=1422 {ECO:0000312|EMBL:ABI49953.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=T-6;
RA Shoham Y., Gat O., Shulami S., Zaide G., Zolotnitsky G., Langut Y.;
RT "Hemicellulose utilization cluster in Geobacillus stearothermophilus strain
RT T-6.";
RL Submitted (JUL-2006) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, INDUCTION, PATHWAY, MUTAGENESIS OF SER-15; ASP-191 AND HIS-194,
RP CATALYTIC MECHANISM, AND ACTIVE SITE.
RC STRAIN=T-6;
RX PubMed=21994937; DOI=10.1074/jbc.m111.301051;
RA Alalouf O., Balazs Y., Volkinshtein M., Grimpel Y., Shoham G., Shoham Y.;
RT "A new family of carbohydrate esterases is represented by a GDSL
RT hydrolase/acetylxylan esterase from Geobacillus stearothermophilus.";
RL J. Biol. Chem. 286:41993-42001(2011).
RN [3]
RP CRYSTALLIZATION.
RC STRAIN=T-6;
RX PubMed=23545652; DOI=10.1107/s1744309113004260;
RA Lansky S., Alalouf O., Solomon V., Alhassid A., Govada L., Chayen N.E.,
RA Chayan N.E., Belrhali H., Shoham Y., Shoham G.;
RT "Crystallization and preliminary crystallographic analysis of Axe2, an
RT acetylxylan esterase from Geobacillus stearothermophilus.";
RL Acta Crystallogr. F 69:430-434(2013).
RN [4]
RP CRYSTALLIZATION, AND MUTAGENESIS OF TYR-184 AND TRP-190.
RC STRAIN=T-6;
RX PubMed=24699743; DOI=10.1107/s2053230x14004129;
RA Lansky S., Alalouf O., Salama R., Dvir H., Shoham Y., Shoham G.;
RT "Preliminary crystallographic analysis of a double mutant of the acetyl
RT xylo-oligosaccharide esterase Axe2 in its dimeric form.";
RL Acta Crystallogr. F 70:476-481(2014).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF MUTANTS ALA-191 AND ALA-194.
RC STRAIN=T-6;
RA Lansky S., Alalouf O., Solomon V., Alhassid A., Belrahli H., Govada L.,
RA Chayan N.E., Shoham Y., Shoham G.;
RT "To be published.";
RL Submitted (MAR-2013) to the PDB data bank.
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF WILD-TYPE AND MUTANT ALA-15,
RP SUBUNIT, AND ACTIVE SITE.
RC STRAIN=T-6;
RX PubMed=24531461; DOI=10.1107/s139900471302840x;
RA Lansky S., Alalouf O., Solomon H.V., Alhassid A., Govada L., Chayen N.E.,
RA Belrhali H., Shoham Y., Shoham G.;
RT "A unique octameric structure of Axe2, an intracellular acetyl-
RT xylooligosaccharide esterase from Geobacillus stearothermophilus.";
RL Acta Crystallogr. D 70:261-278(2014).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.93 ANGSTROMS) OF MUTANTS ALA-55 AND ILE-190.
RC STRAIN=T-6;
RA Lansky S., Alalouf O., Solomon H.V., Belrhali H., Shoham Y., Shoham G.;
RL Submitted (JAN-2014) to the PDB data bank.
CC -!- FUNCTION: Acetylxylan esterase involved in the degradation of xylan, a
CC major structural heterogeneous polysaccharide found in plant biomass
CC representing the second most abundant polysaccharide in the biosphere,
CC after cellulose. Cleaves acetyl side groups from the xylose backbone
CC units of the hemicellulolytic polymer xylan and xylo-oligosaccharides.
CC Hydrolyzes about 20%-30% of the available acetyl groups on fully
CC acetylated birch wood xylan. Completely deacetylates xylobiose
CC peracetate (fully acetylated), and is active on both the alpha- and
CC beta-forms of the sugar. Also hydrolyzes fully acetylated methyl-beta-
CC D-xylopyranoside and methyl-beta-D-glucopyranoside, and the synthetic
CC substrates 2-naphthyl acetate, 4-nitrophenyl acetate, 4-
CC methylumbelliferyl acetate, and phenyl acetate.
CC {ECO:0000269|PubMed:21994937}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Deacetylation of xylans and xylo-oligosaccharides.;
CC EC=3.1.1.72; Evidence={ECO:0000269|PubMed:21994937};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=7 mM for phenyl acetate (at pH 6.0 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:21994937};
CC KM=9 mM for 2-naphtyl acetate (at pH 6.0 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:21994937};
CC KM=3 mM for 4-methylumbelliferyl acetate (at pH 6.0 and 30 degrees
CC Celsius) {ECO:0000269|PubMed:21994937};
CC KM=27 mM for 4-nitrophenyl acetate (at pH 6.0 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:21994937};
CC Note=kcat is 77 sec(-1) with phenyl acetate as substrate. kcat is 190
CC sec(-1) with 2-naphtyl acetate as substrate. kcat is 123 sec(-1) with
CC 4-methylumbelliferyl acetate as substrate. kcat is 31 sec(-1) with 4-
CC nitrophenyl acetate as substrate (at pH 6.0 and 30 degrees Celsius).
CC {ECO:0000269|PubMed:21994937};
CC pH dependence:
CC Optimum pH is 7.1-9.2. {ECO:0000269|PubMed:21994937};
CC Temperature dependence:
CC Optimum temperature is 50-60 degrees Celsius. Is stable at 60 degrees
CC Celsius and loses most of its activity at 70 degrees Celsius.
CC {ECO:0000269|PubMed:21994937};
CC -!- PATHWAY: Glycan degradation; xylan degradation.
CC {ECO:0000269|PubMed:21994937}.
CC -!- SUBUNIT: Homooctamer, presenting a unique donut-shaped quaternary
CC structure built of two staggered tetrameric rings. The eight active
CC sites are organized in four closely situated pairs, which face the
CC relatively wide internal cavity. {ECO:0000269|PubMed:24531461}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:21994937}.
CC -!- INDUCTION: Up-regulated by xylose. {ECO:0000269|PubMed:21994937}.
CC -!- SIMILARITY: Belongs to the 'GDSL' lipolytic enzyme family.
CC {ECO:0000305}.
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DR EMBL; DQ868502; ABI49953.1; -; Genomic_DNA.
DR PDB; 3W7V; X-ray; 1.85 A; A/B=1-219.
DR PDB; 4JHL; X-ray; 1.70 A; A/B=1-219.
DR PDB; 4JJ4; X-ray; 2.13 A; A/B=1-219.
DR PDB; 4JJ6; X-ray; 1.80 A; A/B=1-219.
DR PDB; 4JKO; X-ray; 1.90 A; A/B=1-219.
DR PDB; 4OAO; X-ray; 2.05 A; A/B=1-219.
DR PDB; 4OAP; X-ray; 1.93 A; A/B=1-219.
DR PDB; 5BN1; X-ray; 1.60 A; A/B=1-219.
DR PDBsum; 3W7V; -.
DR PDBsum; 4JHL; -.
DR PDBsum; 4JJ4; -.
DR PDBsum; 4JJ6; -.
DR PDBsum; 4JKO; -.
DR PDBsum; 4OAO; -.
DR PDBsum; 4OAP; -.
DR PDBsum; 5BN1; -.
DR AlphaFoldDB; Q09LX1; -.
DR SMR; Q09LX1; -.
DR BRENDA; 3.1.1.72; 623.
DR UniPathway; UPA00114; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0046555; F:acetylxylan esterase activity; IDA:CACAO.
DR GO; GO:0045493; P:xylan catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.40.50.1110; -; 1.
DR InterPro; IPR013830; SGNH_hydro.
DR InterPro; IPR036514; SGNH_hydro_sf.
DR Pfam; PF13472; Lipase_GDSL_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cytoplasm; Hydrolase;
KW Polysaccharide degradation; Serine esterase.
FT CHAIN 1..219
FT /note="Acetylxylan esterase"
FT /id="PRO_0000434101"
FT ACT_SITE 15
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:21994937,
FT ECO:0000305|PubMed:24531461"
FT ACT_SITE 191
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:21994937,
FT ECO:0000305|PubMed:24531461"
FT ACT_SITE 194
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:21994937,
FT ECO:0000305|PubMed:24531461"
FT SITE 63
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:24531461"
FT SITE 92
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:24531461"
FT MUTAGEN 15
FT /note="S->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:21994937"
FT MUTAGEN 184
FT /note="Y->F: Significant reduction in catalytic activity
FT and modification of the quaternary structure as a
FT homodimer; when associated with P-190."
FT /evidence="ECO:0000269|PubMed:24699743"
FT MUTAGEN 190
FT /note="W->P: Significant reduction in catalytic activity
FT and modification of the quaternary structure as a
FT homodimer; when associated with F-184."
FT /evidence="ECO:0000269|PubMed:24699743"
FT MUTAGEN 191
FT /note="D->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:21994937"
FT MUTAGEN 194
FT /note="H->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:21994937"
FT STRAND 8..14
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 15..18
FT /evidence="ECO:0007829|PDB:5BN1"
FT TURN 23..25
FT /evidence="ECO:0007829|PDB:5BN1"
FT STRAND 28..30
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 31..33
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 37..48
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 50..52
FT /evidence="ECO:0007829|PDB:4JHL"
FT STRAND 55..58
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 66..76
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 78..80
FT /evidence="ECO:0007829|PDB:5BN1"
FT STRAND 83..88
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 91..99
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 104..106
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 110..124
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 125..127
FT /evidence="ECO:0007829|PDB:5BN1"
FT STRAND 128..134
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 146..165
FT /evidence="ECO:0007829|PDB:5BN1"
FT STRAND 168..170
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 172..180
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 185..187
FT /evidence="ECO:0007829|PDB:5BN1"
FT STRAND 192..194
FT /evidence="ECO:0007829|PDB:5BN1"
FT HELIX 197..211
FT /evidence="ECO:0007829|PDB:5BN1"
SQ SEQUENCE 219 AA; 24772 MW; A3F18C97F5F69E32 CRC64;
MKIGSGEKLL FIGDSITDCG RARPEGEGSF GALGTGYVAY VVGLLQAVYP ELGIRVVNKG
ISGNTVRDLK ARWEEDVIAQ KPDWVSIMIG INDVWRQYDL PFMKEKHVYL DEYEATLRSL
VLETKPLVKG IILMTPFYIE GNEQDPMRRT MDQYGRVVKQ IAEETNSLFV DTQAAFNEVL
KTLYPAALAW DRVHPSVAGH MILARAFLRE IGFEWVRSR
