2drs
From Proteopedia
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{{STRUCTURE_2drs| PDB=2drs | SCENE= }} | {{STRUCTURE_2drs| PDB=2drs | SCENE= }} | ||
| - | + | ===Crystal structure of reducing-end-xylose releasing exo-oligoxylanase D263S mutant=== | |
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| - | ==Overview== | ||
| - | Reducing end xylose-releasing exooligoxylanase (Rex, EC 3.2.1.156) is an inverting GH that hydrolyzes xylooligosaccharides (> or = X3) to release X1 at their reducing end. The wild-type enzyme exhibited the Hehre resynthesis hydrolysis mechanism, in which alpha-X2F was hydrolyzed to X2 and HF in the presence of X1 as an acceptor molecule. However, the transglycosidation product (X3) was not detectable in the reaction. To convert reducing end xylose-releasing exooligoxylanase to glycosynthase, derivatives with mutations in the catalytic base (Asp-263) were constructed by saturation random mutagenesis. Nine amino acid residue mutants (Asp-263 to Gly, Ala, Val, Thr, Leu, Asn, Cys, Pro, or Ser) were found to possess glycosynthase activity forming X3 from alpha-X2F and X1. Among them, D263C showed the highest level of X3 production, and D263N exhibited the fastest consumption of alpha-X2F. The D263C mutant showed 10-fold lower hydrolytic activity than D263N, resulting in the highest yield of X3. X2 was formed from the early stage of the reaction of the D263C mutant, indicating that a portion of the X3 formed by condensation was hydrolyzed before its release from the enzyme. To acquire glycosynthase activity from inverting enzymes, it is important to minimize the decrease in F(-)-releasing activity while maximizing the decrease in the hydrolytic activity. The present study expands the possibility of conversion of glycosynthases from inverting enzymes. | ||
==About this Structure== | ==About this Structure== | ||
| - | 2DRS is a | + | 2DRS is a 1 chain structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_halodurans Bacillus halodurans]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2DRS OCA]. |
==Reference== | ==Reference== | ||
| - | + | <ref group="xtra">PMID:16301312</ref><ref group="xtra">PMID:15718242</ref><ref group="xtra">PMID:16511021</ref><ref group="xtra">PMID:15491996</ref><references group="xtra"/> | |
[[Category: Bacillus halodurans]] | [[Category: Bacillus halodurans]] | ||
[[Category: Oligosaccharide reducing-end xylanase]] | [[Category: Oligosaccharide reducing-end xylanase]] | ||
| - | [[Category: Single protein]] | ||
[[Category: Fushinobu, S.]] | [[Category: Fushinobu, S.]] | ||
[[Category: Hidaka, M.]] | [[Category: Hidaka, M.]] | ||
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[[Category: Nppsfa]] | [[Category: Nppsfa]] | ||
[[Category: Structural genomic]] | [[Category: Structural genomic]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | |
| + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Feb 18 04:20:13 2009'' | ||
Revision as of 02:20, 18 February 2009
Crystal structure of reducing-end-xylose releasing exo-oligoxylanase D263S mutant
About this Structure
2DRS is a 1 chain structure of sequence from Bacillus halodurans. Full crystallographic information is available from OCA.
Reference
- Honda Y, Kitaoka M. The first glycosynthase derived from an inverting glycoside hydrolase. J Biol Chem. 2006 Jan 20;281(3):1426-31. Epub 2005 Nov 21. PMID:16301312 doi:10.1074/jbc.M511202200
- Fushinobu S, Hidaka M, Honda Y, Wakagi T, Shoun H, Kitaoka M. Structural basis for the specificity of the reducing end xylose-releasing exo-oligoxylanase from Bacillus halodurans C-125. J Biol Chem. 2005 Apr 29;280(17):17180-6. Epub 2005 Feb 17. PMID:15718242 doi:10.1074/jbc.M413693200
- Honda Y, Fushinobu S, Hidaka M, Wakagi T, Shoun H, Kitaoka M. Crystallization and preliminary X-ray analysis of reducing-end xylose-releasing exo-oligoxylanase from Bacillus halodurans C-125. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Mar 1;61(Pt, 3):291-2. Epub 2005 Feb 12. PMID:16511021 doi:10.1107/S1744309105003635
- Honda Y, Kitaoka M. A family 8 glycoside hydrolase from Bacillus halodurans C-125 (BH2105) is a reducing end xylose-releasing exo-oligoxylanase. J Biol Chem. 2004 Dec 31;279(53):55097-103. Epub 2004 Oct 18. PMID:15491996 doi:10.1074/jbc.M409832200
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