Molecular and physiological role of the trehalose-hydrolyzing alpha-glucosidase from Thermus thermophilus HB27.
Trehalose supports the growth of Thermus thermophilus strain HB27, but the absence of obvious genes for the hydrolysis of this disaccharide in the genome led us to search for enzymes for such a purpose. We expressed a putative alpha-glucosidase gene (TTC0107), characterized the recombinant enzyme, and found that the preferred substrate was alpha,alpha-1,1-trehalose, a new feature among alpha-glucosidases. The enzyme could also hydrolyze the disaccharides kojibiose and sucrose (alpha-1,2 linkage), nigerose and turanose (alpha-1,3), leucrose (alpha-1,5), isomaltose and palatinose (alpha-1,6), and maltose (alpha-1,4) to a lesser extent. Trehalose was not, however, a substrate for the highly homologous alpha-glucosidase from T. thermophilus strain GK24. The reciprocal replacement of a peptide containing eight amino acids in the alpha-glucosidases from strains HB27 (LGEHNLPP) and GK24 (EPTAYHTL) reduced the ability of the former to hydrolyze trehalose and provided trehalose-hydrolytic activity to the latter, showing that LGEHNLPP is necessary for trehalose recognition. Furthermore, disruption of the alpha-glucosidase gene significantly affected the growth of T. thermophilus HB27 in minimal medium supplemented with trehalose, isomaltose, sucrose, or palatinose, to a lesser extent with maltose, but not with cellobiose (not a substrate for the alpha-glucosidase), indicating that the alpha-glucosidase is important for the assimilation of those four disaccharides but that it is also implicated in maltose catabolism.
Alarico
Susana
S
Centro de Neurociências e Biologia Celular, Departamento de Zoologia, Universidade de Coimbra, 3004-517 Coimbra, Portugal.
da Costa
Milton S
MS
Empadinhas
Nuno
N
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D004187:Disaccharides / Q000378:metabolism; D007534:Isomaltose / Q000031:analogs & derivatives / Q000378:metabolism; D007700:Kinetics; D008320:Maltose / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics; D014199:Trehalose / Q000378:metabolism*; D000520:alpha-Glucosidases / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004187:Disaccharides; D011994:Recombinant Proteins; C014637:laminaribiose; D013395:Sucrose; D007534:Isomaltose; D008320:Maltose; C029454:turanose; C008237:kojibiose; D014199:Trehalose; D000520:alpha-Glucosidases; C008189:isomaltulose; C060525:leucrose