3WFZ: Crystal Structure Of Galacto-n-biose/lacto-n-biose I Phosphorylase C236y Mutant

Citation:
Abstract
Galacto-N-biose/lacto-N-biose I phosphorylase (GLNBP) is the key enzyme in the enzymatic production of lacto-N-biose I. For the purpose of industrial use, we improved the thermostability of GLNBP by evolutionary engineering in which five substitutions in the amino acid sequence were selected from a random mutagenesis GLNBP library constructed using error-prone polymerase chain reaction. Among them, C236Y and D576V mutants showed considerably improved thermostability. Structural analysis of C236Y revealed that the hydroxyl group of Tyr236 forms a hydrogen bond with the carboxyl group of E319. The C236Y and D576V mutations together contributed to the thermostability. The C236Y/D576V mutant exhibited 20 degrees C higher thermostability than the wild type.
PDB ID: 3WFZDownload
MMDB ID: 114387
PDB Deposition Date: 2013/7/25
Updated in MMDB: 2013/10
Experimental Method:
x-ray diffraction
Resolution: 2.6  Å
Source Organism:
Similar Structures:
Biological Unit for 3WFZ: dimeric; determined by author and by software (PISA)
Molecular Components in 3WFZ
Label Count Molecule
Proteins (2 molecules)
2
Lacto-n-biose Phosphorylase
Molecule annotation
Chemicals (2 molecules)
1
2
* Click molecule labels to explore molecular sequence information.

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