3wx5

From Proteopedia

Crystal structure of metagenome-derived glycoside hydrolase family 12 endoglucanase

Structural highlights

3wx5 is a 2 chain structure with sequence from Uncultured bacterium. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 1.85Å
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

W8PWF3_9BACT

Publication Abstract from PubMed

Ten genes encoding novel cellulases with putative signal peptides at the N-terminus, termed pre-LC-CelA-J, were isolated from a fosmid library of a leaf-branch compost metagenome by functional screening using agar plates containing carboxymethyl cellulose and trypan blue. All the cellulases except pre-LC-CelG have a 14-29 residue long flexible linker (FL) between the signal peptide and the catalytic domain. LC-CelA without a signal peptide (residues 20-261), which shows 76% amino acid sequence identity to Cel12A from Rhodothermus marinus (RmCel12A), was overproduced in Escherichia coli, purified and characterized. LC-CelA exhibited its highest activity across a broad pH range (pH 5-9) and at 90 degrees C, indicating that LC-CelA is a highly thermostable cellulase, like RmCel12A. The crystal structure of LC-CelA was determined at 1.85 A resolution and is nearly identical to that of RmCel12A determined in a form without the FL. Both proteins contain two disulfide bonds. LC-CelA has a 16-residue FL (residues 20-35), most of which is not visible in the electron density map, probably due to structural disorder. However, Glu34 and Pro35 form hydrogen bonds with the central region of the protein. DeltaFL-LC-CelA (residues 36-261) and E34A-LC-CelA with a single Glu34 --> Ala mutation were therefore constructed and characterized. DeltaFL-LC-CelA and E34A-LC-CelA had lower melting temperatures (T m) than LC-CelA by 14.7 and 12.0 degrees C respectively. The T m of LC-CelA was also decreased by 28.0 degrees C in the presence of dithiothreitol. These results suggest that Glu34-mediated hydrogen bonds and the two disulfide bonds contribute to the stabilization of LC-CelA.

Structure and stability of metagenome-derived glycoside hydrolase family 12 cellulase (LC-CelA) a homolog of Cel12A from Rhodothermus marinus.,Okano H, Ozaki M, Kanaya E, Kim JJ, Angkawidjaja C, Koga Y, Kanaya S FEBS Open Bio. 2014 Oct 31;4:936-46. doi: 10.1016/j.fob.2014.10.013. eCollection , 2014. PMID:25426413^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Okano H, Ozaki M, Kanaya E, Kim JJ, Angkawidjaja C, Koga Y, Kanaya S. Structure and stability of metagenome-derived glycoside hydrolase family 12 cellulase (LC-CelA) a homolog of Cel12A from Rhodothermus marinus. FEBS Open Bio. 2014 Oct 31;4:936-46. doi: 10.1016/j.fob.2014.10.013. eCollection , 2014. PMID:25426413 doi:http://dx.doi.org/10.1016/j.fob.2014.10.013