Trehalose catabolism enzymes in L3 and L4 larvae of Anisakis simplex.
The presence of trehalase and trehalose phosphorylase in L3 and L4 larvae of Anisakis simplex was demonstrated. The activity of trehalase and trehalose phosphorylase in L3 larvae was 6 and 10 times higher, respectively, than in L4 larvae. This suggests that trehalose metabolism is more important for L3 than LA larvae. Trehalases of L3 and L4 differ in their characteristics. The enzyme of L3 was present mainly in the lysosomes and cytosol, whereas in L4 the highest enzyme activity was measured in the lysosomal fraction. Trehalase activity was increased by 29% in L3 and 55% in L4 with the addition of Mg2+ (0.1 mmol). Tris inhibited trehalase in L3 larvae by 42% and in L4 by 25%. The enzymes differed in their reaction to EDTA, CaCl2, ZnCl2, and CH2ICOOH (all 0.1 mmol). High activity of trehalase from L3 larvae was measured within the pH range of 5.0 to 6.5, with an optimum pH of 6.1. The trehalase was a thermally tolerant enzyme from 25 C to 60 C. The enzyme lost half of its activity after preincubation without substrate above 75 C. The paper also discusses the similarities and differences in characteristics of trehalase from A. simplex larvae and presents the comparison to enzymes from other nematodes.
Lopieńska-Biernat
E
E
University Warmia and Mazury, Faculty of Biology, Department of Biochemistry, Olsztyn 10-957, Oczapowskiego 1A, Poland. ela.lopienska@uwm.edu.pl
Zółtowska
K
K
Rokicki
J
J
D000818:Animals; D017163:Anisakis / Q000201:enzymology* / Q000648:ultrastructure; D002122:Calcium Chloride / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D019807:Iodoacetic Acid / Q000494:pharmacology; D007814:Larva / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D015636:Magnesium Chloride / Q000494:pharmacology; D013696:Temperature; D014198:Trehalase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014199:Trehalose / Q000378:metabolism*; D014325:Tromethamine / Q000494:pharmacology; D017967:Zinc Compounds / Q000494:pharmacology
D002712:Chlorides; D004791:Enzyme Inhibitors; D017967:Zinc Compounds; D014325:Tromethamine; D015636:Magnesium Chloride; C016837:zinc chloride; D004492:Edetic Acid; D014199:Trehalose; D005964:Glucosyltransferases; C023181:alpha,alpha-trehalose phosphorylase; D014198:Trehalase; D002122:Calcium Chloride; D019807:Iodoacetic Acid