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Characterization of intracellular esterase A from Bacillus subtilis
Biochimica Et Biophysica Acta
|March 11, 1976
Summary
This study purified Esterase A from Bacillus subtilis, finding it effectively hydrolyzes acetate esters but not amino acid esters. Its activity is unaffected by alcohols but inhibited by heavy metals and specific chemical agents.
Area of Science:
- Biochemistry
- Enzymology
- Microbial biochemistry
Background:
- Esterases are crucial enzymes involved in various biological processes.
- Understanding the specific properties of microbial esterases can lead to novel biotechnological applications.
Purpose of the Study:
- To purify and characterize Esterase A from Bacillus subtilis SR22.
- To determine the substrate specificity and inhibition patterns of the purified enzyme.
Main Methods:
- Enzyme purification using differential precipitation, ion-exchange chromatography (DEAE-cellulose), and gel filtration (Bio-Rad P-150).
- Enzyme activity assays with various ester substrates.
- Inhibition studies using different chemical agents and alcohols.
Main Results:
- Esterase A was purified approximately 400-fold with a 59% yield.
- The enzyme efficiently hydrolyzed aliphatic and aromatic acetate esters but not amino acid esters.
- Aliphatic alcohols did not inhibit hydrolysis of p-nitrophenyl acetate; mercuric chloride, diisopropylfluorophosphate, eserine, and sodium fluoride were potent inhibitors.
Conclusions:
- Bacillus subtilis SR22 Esterase A exhibits specific activity towards acetate esters.
- The enzyme's inhibition profile suggests a serine hydrolase mechanism and sensitivity to heavy metals.