PDBsum entry 1v6y

Hydrolase

PDB id: 1v6y

Contents

Protein chain

316 a.a. *

Waters ×240

* Residue conservation analysis

PDB id:

1v6y

Name:

Hydrolase

Title:

Crystal structure of chimeric xylanase between streptomyces olivaceoviridis e-86 fxyn and cellulomonas fimi cex

Structure:

Beta-xylanase,exoglucanase/xylanase. Chain: a. Engineered: yes. Other_details: chimeric enzyme from two xylanases,chimeric enzyme from two xylanases

Source:

Streptomyces olivaceoviridis, cellulomonas fimi. Organism_taxid: 1921, 1708. Gene: cex, xynb. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.20Å R-factor: 0.153 R-free: 0.198

Authors:

S.Kaneko,H.Ichinose,Z.Fujimoto,A.Kuno,K.Yura,M.Go,H.Mizuno, I.Kusakabe,H.Kobayashi

Key ref:

S.Kaneko et al. (2004). Structure and function of a family 10 beta-xylanase chimera of Streptomyces olivaceoviridis E-86 FXYN and Cellulomonas fimi Cex. J Biol Chem, 279, 26619-26626. PubMed id: 15078885 DOI: 10.1074/jbc.M308899200

Date:

04-Dec-03 Release date: 07-Sep-04

Protein chain

Q7SI98  (Q7SI98_STROI) -  Beta-xylanase from Streptomyces olivaceoviridis

Seq: 436 a.a.

Struc: 316 a.a.*

* PDB and UniProt seqs differ at 69 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.2.1.8 - endo-1,4-beta-xylanase.
• Reaction: Endohydrolysis of 1,4-beta-D-xylosidic linkages in xylans.

DOI no: 10.1074/jbc.M308899200

J Biol Chem 279:26619-26626 (2004)

PubMed id: 15078885

Structure and function of a family 10 beta-xylanase chimera of Streptomyces olivaceoviridis E-86 FXYN and Cellulomonas fimi Cex.

S.Kaneko, H.Ichinose, Z.Fujimoto, A.Kuno, K.Yura, M.Go, H.Mizuno, I.Kusakabe, H.Kobayashi.

ABSTRACT

The catalytic domain of xylanases belonging to glycoside hydrolase family 10 (GH10) can be divided into 22 modules (M1 to M22; Sato, Y., Niimura, Y., Yura, K., and Go, M. (1999) Gene (Amst.) 238, 93-101). Inspection of the crystal structure of a GH10 xylanase from Streptomyces olivaceoviridis E-86 (SoXyn10A) revealed that the catalytic domain of GH10 xylanases can be dissected into two parts, an N-terminal larger region and C-terminal smaller region, by the substrate binding cleft, corresponding to the module border between M14 and M15. It has been suggested that the topology of the substrate binding clefts of GH10 xylanases are not conserved (Charnock, S. J., Spurway, T. D., Xie, H., Beylot, M. H., Virden, R., Warren, R. A. J., Hazlewood, G. P., and Gilbert, H. J. (1998) J. Biol. Chem. 273, 32187-32199). To facilitate a greater understanding of the structure-function relationship of the substrate binding cleft of GH10 xylanases, a chimeric xylanase between SoXyn10A and Xyn10A from Cellulomonas fimi (CfXyn10A) was constructed, and the topology of the hybrid substrate binding cleft established. At the three-dimensional level, SoXyn10A and CfXyn10A appear to possess 5 subsites, with the amino acid residues comprising subsites -3 to +1 being well conserved, although the +2 subsites are quite different. Biochemical analyses of the chimeric enzyme along with SoXyn10A and CfXyn10A indicated that differences in the structure of subsite +2 influence bond cleavage frequencies and the catalytic efficiency of xylooligosaccharide hydrolysis. The hybrid enzyme constructed in this study displays fascinating biochemistry, with an interesting combination of properties from the parent enzymes, resulting in a low production of xylose.

Selected figure(s)

Figure 5.
FIG. 5. Bond cleavage frequencies of xylooligosaccharides by CfXyn10A, SoXyn10A, and FC-14-15.

Figure 7.
FIG. 7. HPAEC-PAD analysis of soluble birchwood xylan hydrolysis by CfXyn10A, SoXyn10A, and FC-14-15. Birchwood xylan hydrolysate by SoXyn10A (A), CfXyn10A (B), and FC-14-15 (C) were applied to the HPAEC-PAD system. The positions at which xylose (a), xylobiose (b), xylotriose (c), xylotetraose (d), and xylooligosaccharides substituted by 4-O-methyl glucuronic acid (e) were eluted from the HPAEC column are indicated.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2004, 279, 26619-26626) copyright 2004.

Figures were selected by an automated process.