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Contents |
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* Residue conservation analysis
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PDB id:
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Hydrolase
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Title:
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Crystal structure of a glycoside hydrolase family 44 endoglu produced by clostridium acetobutylium atcc 824
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Structure:
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Endoglucanase a. Chain: a. Fragment: unp residues 34-541(fragment excluding signal pep dockerin domain). Synonym: endo-1,4-beta-glucanase, cellulase a. Engineered: yes
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Source:
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Clostridium acetobutylicum. Organism_taxid: 272562. Strain: atcc 824. Gene: cac0915, ca_c0915. Expressed in: escherichia coli. Expression_system_taxid: 469008.
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Resolution:
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2.20Å
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R-factor:
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0.155
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R-free:
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0.238
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Authors:
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C.D.Warner,J.A.Hoy,C.F.Ford,R.B.Honzatko,P.J.Reilly
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Key ref:
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C.D.Warner
et al.
(2010).
Tertiary structure and characterization of a glycoside hydrolase family 44 endoglucanase from Clostridium acetobutylicum.
Appl Environ Microbiol,
76,
338-346.
PubMed id:
DOI:
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Date:
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05-Aug-09
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Release date:
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18-Aug-09
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PROCHECK
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Headers
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References
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Q977Y3
(Q977Y3_CLOAB) -
Endoglucanase A (Endo-1,4-beta-glucanase) (Cellulase A), secreted; dockerin domain
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Seq:
Struc:
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606 a.a.
512 a.a.*
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Key: |
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PfamA domain |
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Secondary structure |
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*
PDB and UniProt seqs differ
at 4 residue positions (black
crosses)
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DOI no:
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Appl Environ Microbiol
76:338-346
(2010)
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PubMed id:
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Tertiary structure and characterization of a glycoside hydrolase family 44 endoglucanase from Clostridium acetobutylicum.
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C.D.Warner,
J.A.Hoy,
T.C.Shilling,
M.J.Linnen,
N.D.Ginder,
C.F.Ford,
R.B.Honzatko,
P.J.Reilly.
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ABSTRACT
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A gene encoding a glycoside hydrolase family 44 (GH44) protein from Clostridium
acetobutylicum ATCC 824 was synthesized and transformed into Escherichia coli.
The previously uncharacterized protein was expressed with a C-terminal His tag
and purified by nickel-nitrilotriacetic acid affinity chromatography.
Crystallization and X-ray diffraction to a 2.2-A resolution revealed a triose
phosphate isomerase (TIM) barrel-like structure with additional Greek key and
beta-sandwich folds, similar to other GH44 crystal structures. The enzyme
hydrolyzes cellotetraose and larger cellooligosaccharides, yielding an
unbalanced product distribution, including some glucose. It attacks
carboxymethylcellulose and xylan at approximately the same rates. Its activity
on carboxymethylcellulose is much higher than that of the isolated C.
acetobutylicum cellulosome. It also extensively converts lichenan to
oligosaccharides of intermediate size and attacks Avicel to a limited extent.
The enzyme has an optimal temperature in a 10-min assay of 55 degrees C and an
optimal pH of 5.0.
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Links
