Determination of trehalose in biological samples by a simple and stable trehalase preparation.
A three step purification procedure for trehalase from Saccharomyces cerevisiae with a recovery of 76% of the original activity is presented. The enzyme was activated by a heat shock treatment prior to homogenization of the cells. A mutant strain deleted in SUC genes was used to avoid contamination by invertase. The lyophylized enzyme was stable for, at least, 5 months and could be used to determine trehalose in the range 25 to 500 nmol. The preparation was free of inspecific phosphatases allowing for trehalose determinations in yeast cell free extracts and in insect hemolymph.
Araujo
P S
PS
Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Brazil.
Panek
A C
AC
Ferreira
R
R
Panek
A D
AD
D002845:Chromatography; D004187:Disaccharides / Q000032:analysis*; D004789:Enzyme Activation; D006360:Heat-Shock Proteins / Q000302:isolation & purification; D011094:Polyethyleneimine; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D014198:Trehalase / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D014199:Trehalose / Q000032:analysis*
D004187:Disaccharides; D006360:Heat-Shock Proteins; D011094:Polyethyleneimine; D014199:Trehalose; D014198:Trehalase