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Some properties of monkey intestinal sucrase
1Department of Life Sciences, University of Bombay, Vidyanagari, Santacruz.
Indian Journal of Biochemistry & Biophysics
|April 1, 1990
Summary
Researchers purified monkey small intestine sucrase, revealing a hetero-dimeric enzyme with distinct maltase and sucrase active sites. This purification provides a homogeneous enzyme for further biochemical studies.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Biology
Background:
- Sucrase is a key enzyme in carbohydrate digestion found in the small intestine.
- Understanding sucrase structure and function is crucial for comprehending nutrient absorption and related disorders.
Purpose of the Study:
- To purify and characterize sucrase from monkey small intestine.
- To investigate the catalytic properties and subunit composition of the purified enzyme.
Main Methods:
- Enzyme purification using detergent solubilization and gel electrophoresis.
- Determination of molecular weight via gel filtration.
- Analysis of enzyme subunits using SDS-PAGE.
- Enzyme kinetics and inhibition studies (mixed substrate, PCMB, Tris).
- Immunological characterization using polyclonal antiserum.
Main Results:
- Sucrase was purified 388-fold to homogeneity with 36% recovery.
- The enzyme's molecular weight was determined to be 263 kDa.
- SDS-PAGE indicated a hetero-dimeric structure.
- Kinetic studies suggested two active sites: one for maltase and one for sucrase, with shared isomaltase activity.
- Antiserum confirmed the purity and antigenicity of the enzyme.
Conclusions:
- Monkey small intestine sucrase is a hetero-dimer with distinct catalytic sites.
- The purified enzyme serves as a valuable tool for studying carbohydrate digestion and enzyme mechanisms.