Summary
A soluble trehalase was purified more than 200-fold from the male accessory gland of the American cockroach,Periplaneta americana, by CM-cellulose, hydrophobic chromatography, and Sephacryl S-200 gel filtration. The final preparation was homogeneous as judged by polyacryl-amide gel electrophoresis in the absence and presence of SDS, isoelectric focusing, and immuno-diffusion tests. The purified enzyme was maximally active at pH 5.2, and showed high specificity for trehalose with aK m of 0.98 mM. The isoelectric point was 4.7. The molecular weight of the enzyme (75,000) was determined by molecular sieve chromatography and SDS-polyacrylamide gel electrophoresis. The amino acid composition was determined and compared with those of trehalases purified from other sources. The trehalase could be stained for carbohydrate with the periodic acid-Schiff's reagent following SDS-polyacrylamide gel electrophoresis, indicating that it was a glycoprotein. Another soluble trehalase and two types of fat body trehalases could be highly purified by the method described. A comparison of the properties of trehalases from the accessory gland and the fat body showed some resemblance.
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Abbreviations
- CM :
-
carboxymethyl
- DEAE :
-
diethylaminoethyl
- SDS :
-
sodium dodecyl sulfate
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Ogiso, M., Shinohara, Y., Hanaoka, K. et al. Further purification and characterization of trehalases from the American cockroach,Periplaneta americana . J Comp Physiol B 155, 553–560 (1985). https://doi.org/10.1007/BF00694444
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DOI: https://doi.org/10.1007/BF00694444