RRC ID 55159
著者 Wang B, Ji SQ, Ma XQ, Lu M, Wang LS, Li FL.
タイトル Substitution of one calcium-binding amino acid strengthens substrate binding in a thermophilic alginate lyase.
ジャーナル FEBS Lett
Abstract Ligand binding is sensitive to temperatures since noncovalent bonds between the binding site and ligand could be broken by heat. How metal ion-binding amino acids in alginate lyase evolve to achieve tight substrate binding in a hostile environment remains unknown. An endolytic alginate lyase AlgAT0 specifically cleaved the M-G glycosidic bond and released disaccharides as the main end product. Four conserved calcium-binding sites were predicted and the supplement of Ca2+ led to enhanced substrate binding and protein stability. Among the four conserved calcium-binding sites, one substitution of aspartate for glutamate in AlgAT0 was proved to stimulate Ca2+ affinity. This study suggested that substrate affinity of polysaccharide lyases could be improved by tight binding to Ca2+ via one amino acid substitution.
巻・号 592(3)
ページ 369-379
公開日 2018-2-1
DOI 10.1002/1873-3468.12965
PMID 29292503
MeSH Amino Acid Substitution* Binding Sites Calcium / metabolism* Calorimetry, Differential Scanning Circular Dichroism Disaccharides / metabolism Models, Molecular Polysaccharide-Lyases / chemistry* Polysaccharide-Lyases / genetics Polysaccharide-Lyases / metabolism* Protein Conformation Substrate Specificity Thermodynamics
IF 2.675
引用数 2
リソース情報
一般微生物 JCM 30481