ALPHA,ALPHA-TREHALOSE GLUCOHYDROLASE

Uses

Trehalase has been used:

• in a study to assess changes in carbohydrate metabolism in Plasmopara viticola-infected grapevine leaves
• in a study to investigate growth arrest by trehalose-6-phosphate
• to quantify the intracellular trehalose in Lactobacillus casei cells

General Description

Trehalase is a glycosidase that occurs in many forms and is found in all tissues. It belongs to the glycoside hydrolase 37 (GH37) family of the carbohydrate-active enzyme (CAZy) classification.

Biochem/physiol Actions

Trehalase catalyzes the hydrolysis of trehalose to produce two glucose units. It mediates flight metabolism, molting induced chitin production and tolerance towards cold temperature.

Purification Methods

This trehalase is purified by solubilising in Triton X-100 and sodium deoxycholate, and submitting to gel filtration, ion-exchange chromatography, conA-Sepharose chromatography, phenyl-Sepharose CL-4B hydrophobic interaction chromatography, Tris-Sepharose 6B affinity and hydrolyapatite chromatography. Activity is increased 3000-fold. [Yoneyama Arch Biochem Biophys 255 168 1987.] T4-RNA ligase (from bacteriophage-infected E.coli ) Mr 43,500, [EC 6.5.1.3 for RNA lyase]. This ligase is purified by differential centrifugation and separation on a Sephadex A-25 column, then through hydroxylapatite and DEAE-glycerol using Aff-Gel Blue to remove DNAase activity. (Greater than 90% of the protein in the enzyme preparation migrated as a single band on gradient polyacrylamide gels containing SDS during electrophoresis.) [McCoy et al. Biochim Biophys Acta 562 149 1979.]