Partial purification and characterization of trehalase from axenically grown myxamoebae of Dictyostelium discoideum

Jyothi Gupta; David Allen Cotter

doi:10.1016/0304-4165(90)90085-B

Partial purification and characterization of trehalase from axenically grown myxamoebae of Dictyostelium discoideum

Jyothi Gupta, David Allen Cotter

Research output: Contribution to journal › Article › peer-review

Abstract

Lysosomal trehalase from the myxamoebae of Dictyostelium discoideum has been partially purified. The behavior of the enzyme under different chromatographic and electrophoretic conditions reveals its close similarities to other lysosomal enzymes that have been studied earlier. The cellular trehalase, which is electrophoretically homogenous, appears as two peaks of activity when subjected to hydroxypatite and gel filtration chromatography. The enzyme has isoelectric points of 4.0 and < 2.5. Among natural disaccharides tested, the purified trehalase showed absolute specificity for trehalose with an apparent K_m of 1.15 mM. However, the enzyme efficiently utilized the synthetic sugar α-d-glucosyl fluoride as a substrate. Various methods were employed to estimate the apparent molecular weight, which was found to lie in the range of 30-162 kDa.

Original language	English (US)
Pages (from-to)	243-248
Number of pages	6
Journal	BBA - General Subjects
Volume	1035
Issue number	3
DOIs	https://doi.org/10.1016/0304-4165(90)90085-B
State	Published - Sep 14 1990
Externally published	Yes

Keywords

(D. discoideum)
Glycoprotein
Lysosomal enzmye
Trehalase

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/0304-4165(90)90085-B

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@article{3023c8328f474c3a84be5f276f07ffb6,

title = "Partial purification and characterization of trehalase from axenically grown myxamoebae of Dictyostelium discoideum",

abstract = "Lysosomal trehalase from the myxamoebae of Dictyostelium discoideum has been partially purified. The behavior of the enzyme under different chromatographic and electrophoretic conditions reveals its close similarities to other lysosomal enzymes that have been studied earlier. The cellular trehalase, which is electrophoretically homogenous, appears as two peaks of activity when subjected to hydroxypatite and gel filtration chromatography. The enzyme has isoelectric points of 4.0 and < 2.5. Among natural disaccharides tested, the purified trehalase showed absolute specificity for trehalose with an apparent Km of 1.15 mM. However, the enzyme efficiently utilized the synthetic sugar α-d-glucosyl fluoride as a substrate. Various methods were employed to estimate the apparent molecular weight, which was found to lie in the range of 30-162 kDa.",

keywords = "(D. discoideum), Glycoprotein, Lysosomal enzmye, Trehalase",

author = "Jyothi Gupta and Cotter, {David Allen}",

note = "Funding Information: We thank Dr. E.J. Hehre, Department of Microbiology and Immunology, Albert Einstein School of Medicine, for the geneorus gift of a-D-glucosyl fluoride and Dr. K.E. Taylor of the University of Windsor for his help with NMR and kinetic studies. This work was supported, in part, by grants from the Natural Sciences and Engineering Research Council of Canada.",

year = "1990",

month = sep,

day = "14",

doi = "10.1016/0304-4165(90)90085-B",

language = "English (US)",

volume = "1035",

pages = "243--248",

journal = "BBA - General Subjects",

issn = "0304-4165",

publisher = "Elsevier B.V.",

number = "3",

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TY - JOUR

T1 - Partial purification and characterization of trehalase from axenically grown myxamoebae of Dictyostelium discoideum

AU - Gupta, Jyothi

AU - Cotter, David Allen

N1 - Funding Information: We thank Dr. E.J. Hehre, Department of Microbiology and Immunology, Albert Einstein School of Medicine, for the geneorus gift of a-D-glucosyl fluoride and Dr. K.E. Taylor of the University of Windsor for his help with NMR and kinetic studies. This work was supported, in part, by grants from the Natural Sciences and Engineering Research Council of Canada.

PY - 1990/9/14

Y1 - 1990/9/14

N2 - Lysosomal trehalase from the myxamoebae of Dictyostelium discoideum has been partially purified. The behavior of the enzyme under different chromatographic and electrophoretic conditions reveals its close similarities to other lysosomal enzymes that have been studied earlier. The cellular trehalase, which is electrophoretically homogenous, appears as two peaks of activity when subjected to hydroxypatite and gel filtration chromatography. The enzyme has isoelectric points of 4.0 and < 2.5. Among natural disaccharides tested, the purified trehalase showed absolute specificity for trehalose with an apparent Km of 1.15 mM. However, the enzyme efficiently utilized the synthetic sugar α-d-glucosyl fluoride as a substrate. Various methods were employed to estimate the apparent molecular weight, which was found to lie in the range of 30-162 kDa.

AB - Lysosomal trehalase from the myxamoebae of Dictyostelium discoideum has been partially purified. The behavior of the enzyme under different chromatographic and electrophoretic conditions reveals its close similarities to other lysosomal enzymes that have been studied earlier. The cellular trehalase, which is electrophoretically homogenous, appears as two peaks of activity when subjected to hydroxypatite and gel filtration chromatography. The enzyme has isoelectric points of 4.0 and < 2.5. Among natural disaccharides tested, the purified trehalase showed absolute specificity for trehalose with an apparent Km of 1.15 mM. However, the enzyme efficiently utilized the synthetic sugar α-d-glucosyl fluoride as a substrate. Various methods were employed to estimate the apparent molecular weight, which was found to lie in the range of 30-162 kDa.

KW - (D. discoideum)

KW - Glycoprotein

KW - Lysosomal enzmye

KW - Trehalase

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SN - 0304-4165

VL - 1035

SP - 243

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JF - BBA - General Subjects

IS - 3

ER -

Partial purification and characterization of trehalase from axenically grown myxamoebae of Dictyostelium discoideum

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Keywords

ASJC Scopus subject areas

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