3WPY: Microbacterium Saccharophilum K-1 Beta-fructofuranosidase Mutant T47s/s200t/f447v/p500s

Citation:
Abstract
A beta-fructofuranosidase from Microbacterium saccharophilum K-1 (formerly known as Arthrobacter sp. K-1) is useful for producing the sweetener lactosucrose (4(G)-beta-D-galactosylsucrose). Thermostability of the beta-fructofuranosidase was enhanced by random mutagenesis and saturation mutagenesis. Clones with enhanced thermostability included mutations at residues Thr47, Ser200, Phe447, Phe470, and Pro500. In the highest stability mutant, T47S/S200T/F447P/F470Y/P500S, the half-life at 60 degrees C was 182 min, 16.5-fold longer than the wild-type enzyme. A comparison of the crystal structures of the full-length wild-type enzyme and three mutants showed that various mechanisms appear to be involved in thermostability enhancement. In particular, the replacement of Phe447 with Val or Pro induced a conformational change in an adjacent residue His477, which results in the formation of a new hydrogen bond in the enzyme. Although the thermostabilization mechanisms of the five residue mutations were explicable on the basis of the crystal structures, it appears to be difficult to predict which amino acid residues should be modified to obtain thermostabilized enzymes.
PDB ID: 3WPYDownload
MMDB ID: 118174
PDB Deposition Date: 2014/1/17
Updated in MMDB: 2014/08
Experimental Method:
x-ray diffraction
Resolution: 2  Å
Source Organism:
Similar Structures:
Biological Unit for 3WPY: monomeric; determined by author and by software (PISA)
Molecular Components in 3WPY
Label Count Molecule
Protein (1 molecule)
1
Beta-fructofuranosidase
Molecule annotation
* Click molecule labels to explore molecular sequence information.

Citing MMDB
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