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Extracellular poly(alpha-L-guluronate)lyase from Corynebacterium sp.: purification, characteristics, and
Y Matsubara1, R Kawada, K Iwasaki
1Kagawa Prefectural Fermentation and Food Experimental Station, Uchinomi, Japan.
Journal of Protein Chemistry
|March 10, 1998
Summary
This study purified alginate lyase from Corynebacterium sp., revealing its structure-function relationship. The enzyme, a poly(alpha-L-guluronate)lyase, showed optimal activity at pH 7.0 and 55°C, with beta-sheets as its common conformation.
Area of Science:
- Biochemistry
- Enzymology
- Marine Biotechnology
Background:
- Alginate lyases are crucial enzymes for alginate degradation.
- Understanding their structure-function relationship is key for biotechnological applications.
- Marine microorganisms are a rich source of novel alginate lyases.
Purpose of the Study:
- To purify and characterize extracellular alginate lyase from Corynebacterium sp.
- To elucidate the structure-function relationship of the purified alginate lyase.
- To investigate the conformational properties and stability of the enzyme.
Main Methods:
- Enzyme purification from culture supernatant.
- Biochemical characterization (molecular mass, isoelectric point, optimal pH/temperature).
- Spectroscopic analysis (SDS-PAGE, gel filtration, circular dichroism).
Main Results:
- Purified alginate lyase (27 kDa) with an isoelectric point of 7.3.
- Optimal activity at pH 7.0 and 55°C; activity enhanced by MnCl2 and NiCl2.
- Identified as endolytic poly(alpha-L-guluronate)lyase, showing beta-form conformation via circular dichroism.
Conclusions:
- The purified alginate lyase exhibits characteristics of poly(alpha-L-guluronate)lyase.
- Beta-sheets appear to be a common structural feature in alginate lyases.
- Enzyme activity and structure are sensitive to denaturants like urea but can be restored.