Purification and partial characterization of an extracellular alginate lyase from Aspergillus oryzae isolated from brown seaweed
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The extracellular enzyme alginate lyase produced from marine fungus Aspergillus oryzae isolated from brown alga Dictyota dichotoma was purified, partially characterized, and evaluated for its sodium alginate depolymerization abilities. The enzyme characterization studies have revealed that alginate lyase consisted of two polypeptides with about 45 and 50 kDa each on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and showed 140-fold higher activity than crude enzyme under optimized pH (6.5) and temperature (35°C) conditions. Zn2+, Mn2+, Cu2+, Mg2+, Co2+ and NaCl were found to enhance the enzyme activity while (Ca2+, Cd2+, Fe2+, Hg2+, Sr2+, Ni2+), glutathione, and metal chelators (ethylenediaminetetraacetic acid and ethylene glycol tetraacetic acid) suppressed the activity. Fourier transform infrared and thin-layer chromatography analysis of depolymerized sodium alginate indicated the enzyme specificity for cleaving at the β-1,4 glycosidic bond between polyM and polyG blocks of sodium alginate and therefore resulted in estimation of relatively higher polyM content than polyG. Comparison of chemical shifts in 13C nuclear magnetic resonance spectra of both polyM and polyG from that of sodium alginate also showed further evidence for enzymatic depolymerization of sodium alginate.
KeywordsAlginate lyase Aspergillus oryzae Fungus PolyM PolyG Sodium alginate
The financial support received from the Council of Scientific and Industrial Research (NWP 018), New Delhi, is gratefully acknowledged. We would also like to thank Mr. Vishal J. Gohel for his assistance in the preparation of the manuscript.
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