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[NAD-dependent formate dehydrogenase from methylotrophic bacteria. Isolation and properties]
Biokhimiia (Moscow, Russia)
|October 1, 1977
Summary
NAD-dependent formate dehydrogenase (FDH) was purified from Bacterium sp 1. The enzyme is stable in the presence of reducing agents like mercaptoethanol.
Area of Science:
- Biochemistry
- Enzymology
- Microbiology
Context:
- Methylotrophic bacteria utilize C1 compounds.
- Formate dehydrogenase (FDH) plays a key role in C1 metabolism.
- Enzyme purification is crucial for biochemical characterization.
Purpose:
- To isolate and purify NAD-dependent formate dehydrogenase (FDH) from a methylotrophic bacterial strain.
- To characterize the homogeneity and stability of the purified enzyme.
Summary:
- NAD-dependent formate dehydrogenase (FDH) was purified from Bacterium sp 1 using ammonium sulfate fractionation, ion-exchange chromatography, and preparative isotachophoresis.
- The purified FDH exhibited homogeneity under analytical polyacrylamide gel electrophoresis and ultracentrifugation, with a sedimentation coefficient of 4.9S.
- Michaelis constants were determined as 1.1 x 10(-4) M for NAD and 1.5 x 10(-2) M for formate. The enzyme is unstable without sulfhydryl compounds but stable in the presence of mercaptoethanol or dithiothreitol.
Impact:
- Provides a purified enzyme for further mechanistic studies.
- Characterization data aids in understanding FDH function in methylotrophs.
- Establishes optimal conditions for enzyme stability and handling.