Purification and characterization of acid trehalase from muscle of Ascaris suum (Nematoda).
Acid trehalase (EC 3.2.1.28) was isolated from muscle of Ascaris suum by fractionating with ammonium sulfate, acetone and column chromatography on DEAE-cellulose and phenyl sepharose CL-4B. The purified homogeneous preparation of native acid trehalase exhibited a molecular mass of 76 kDa and of 38 kDa on SDS-PAGE. The enzyme has the optimum pH 4.9, pI 4.3, Km of 6.6 mM and Vmax=34.5 nM min(-1) x mg(-1). Besides trehalose, it hydrolyses sucrose, isomaltose and maltose and, to a lesser degree melezitose, and it does not act on cellobiose and lactose. Acid trehalase was activated by MgCl2, KNO3, NaCl, CaCl2, CH2ICOOH and p-chloromercuribenzoate and inhibited by EDTA, ZnSO4 and FeCl3.
Dmitryjuk
Małgorzata
M
Department of Biochemistry, Faculty of Biology, University of Warmia and Mazury, Oczapowskiego 1A, Olsztyn 10-957, Poland. dmitm@matman.uwm.edu.pl
Zółtowska
Krystyna
K
D000818:Animals; D017165:Ascaris suum / Q000201:enzymology*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors; D005260:Female; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D012492:Salts; D013379:Substrate Specificity; D013696:Temperature; D014198:Trehalase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D004791:Enzyme Inhibitors; D012492:Salts; D014198:Trehalase