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beta-D-Mannosidase from human placenta: properties and partial purification.
Hoppe-Seyler'S Zeitschrift Fur Physiologische Chemie
|December 1, 1983
Summary
Human placental beta-D-mannosidase was purified 108-fold. Enzyme stability and kinetic parameters were investigated, with purification hampered by activity loss using conventional methods.
Area of Science:
- Biochemistry
- Enzymology
- Human Physiology
Background:
- Beta-D-mannosidase is a key enzyme involved in glycoprotein metabolism.
- Understanding its properties is crucial for diagnosing and potentially treating certain genetic disorders.
- Human placenta is a rich source of various enzymes, including beta-D-mannosidase.
Purpose of the Study:
- To partially purify beta-D-mannosidase from human placenta.
- To investigate the enzyme's stability and the effects of various compounds on its activity.
- To determine key physico-chemical and kinetic parameters of the purified enzyme.
Main Methods:
- Partial purification using ammonium sulfate precipitation.
- Concanavalin A-Sepharose affinity chromatography for enzyme isolation.
- Gel filtration and hydroxylapatite chromatography for further purification.
- Activity assays to monitor enzyme levels and stability.
Main Results:
- Achieved a 108-fold purification of beta-D-mannosidase activity.
- Identified challenges in further purification due to enzyme activity loss with conventional methods.
- Initiated characterization of enzyme stability and kinetic properties.
Conclusions:
- Partial purification of human placental beta-D-mannosidase was successful.
- Enzyme stability and sensitivity to certain compounds were noted.
- Further characterization of physico-chemical and kinetic parameters is warranted.