Discovery of a Kojibiose Hydrolase by Analysis of Specificity-Determining Correlated Positions in Glycoside Hydrolase Family 65
| Autor: | Emma De Beul, Jorick Franceus, Tom Desmet, Alana Jongbloet |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Kojibiose
BIOCHEMICAL-CHARACTERIZATION Amino Acid Motifs Pharmaceutical Science CMA Disaccharides medicine.disease_cause MALTOSE PHOSPHORYLASE Substrate Specificity Analytical Chemistry chemistry.chemical_compound QD241-441 SUBSITE AFFINITIES Drug Discovery Glycoside hydrolase Clade Phylogeny chemistry.chemical_classification Phylogenetic tree Glycoside Hydrolase Family 65 kojibiose glucohydrolase SUBSTRATE-SPECIFICITY Chemistry Chemistry (miscellaneous) correlated positions Molecular Medicine ASPERGILLUS-NIDULANS glycoside phosphorylases glycoside Phosphorylases specificity determinants Computational biology hydrolases VACUOLAR ACID TREHALASE Biology Article Hydrolase Escherichia coli medicine BACILLUS-STEAROTHERMOPHILUS SK-1 Physical and Theoretical Chemistry Mucilaginibacter mallensis Bacteroidetes LACTOBACILLUS-ACIDOPHILUS NCFM Organic Chemistry kojibiase Substrate (chemistry) Biology and Life Sciences ENZYMATIC-PROPERTIES CRYSTALLINE ALPHA-GLUCOSIDASE Enzyme chemistry glycoside hydrolases |
| Zdroj: | Molecules Volume 26 Issue 20 MOLECULES Molecules, Vol 26, Iss 6321, p 6321 (2021) |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules26206321 |
| Popis: | The Glycoside Hydrolase Family 65 (GH65) is an enzyme family of inverting α-glucoside phosphorylases and hydrolases that currently contains 10 characterized enzyme specificities. However, its sequence diversity has never been studied in detail. Here, an in-silico analysis of correlated mutations was performed, revealing specificity-determining positions that facilitate annotation of the family’s phylogenetic tree. By searching these positions for amino acid motifs that do not match those found in previously characterized enzymes from GH65, several clades that may harbor new functions could be identified. Three enzymes from across these regions were expressed in E. coli and their substrate profile was mapped. One of those enzymes, originating from the bacterium Mucilaginibacter mallensis, was found to hydrolyze kojibiose and α-1,2-oligoglucans with high specificity. We propose kojibiose glucohydrolase as the systematic name and kojibiose hydrolase or kojibiase as the short name for this new enzyme. This work illustrates a convenient strategy for mapping the natural diversity of enzyme families and smartly mining the ever-growing number of available sequences in the quest for novel specificities. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|