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Purification and Characterization of Agarase from Marine Bacteria Acinetobacter sp. PS12B and Its Use for Preparing Bioactive Hydrolysate from Agarophyte Red Seaweed Gracilaria verrucosa.

Leema Roseline, T. and Sachindra, N. M. (2018) Purification and Characterization of Agarase from Marine Bacteria Acinetobacter sp. PS12B and Its Use for Preparing Bioactive Hydrolysate from Agarophyte Red Seaweed Gracilaria verrucosa. Applied Biochemistry and Biotechnology, 186. pp. 66-84. ISSN 0273-2289

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Abstract

Acinetobacter strain PS12B was isolated from marine sediment and was found to be a good candidate to degrade agar and produce agarase enzyme. The extracellular agarase enzyme from strain PS12B was purified by ammonium sulfate precipitation followed by DEAE-cellulose ion-exchange chromatography. The specific activity of the crude enzyme which was 1.52 U increased to 45.76 U, after two-stage purification, with an enzyme yield of 9.76%. Purified enzyme had a molecular mass of 24 kDa. The optimum pH and temperature for activity of purified agarase were found to be 8.0 and 40 °C, respectively. The Km and Vmax values for agarase were 4.69 mg/ml and 0.5 μmol/min, respectively. Treatment with EDTA reduced the agarase activity by 58% at 5 mM concentration. The enzyme activity was stimulated by the presence of Fe2+, Mn2+, and Ca2+ ions while reducing reagents (β-mercaptoethanol and dithiothreitol, DTT) enhanced its activity by 30–40%. The purified agarase exhibited tolerance to both detergents and organic solvents. Major hydrolysis products of agar were DP4 and also a mixture of longer oligosaccharides DP6 and DP7. The enzyme hydrolysed seaweed (Gracilaria verrucosa) exhibited strong antioxidant activity in vitro. Successful hydrolysis of seaweed indicates the potential use of the enzyme to produce seaweed hydrolysate having health benefits as well as the industrial application like the production of biofuels.

Item Type: Article
Uncontrolled Keywords: Acinetobacter . Agarase . Degree of polymerization (DP) . Gracilaria verrucosa . Oligosaccharide . Antioxidant activity
Subjects: 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology > 01 Algae
600 Technology > 08 Food technology > 16 Nutritive value > 10 Antioxidant Property
Divisions: Meat Fish and Poultry Technology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 02 Jan 2019 11:15
Last Modified: 02 Jan 2019 11:15
URI: http://ir.cftri.com/id/eprint/13876

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