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Purification and characterization of two agarases from Agarivorans albus OAY02

Paper ID Volume ID Publish Year Pages File Format Full-Text
34536 45032 2014 8 PDF Available
Title
Purification and characterization of two agarases from Agarivorans albus OAY02
Abstract

•Two β-agarases were purified from Agarivorans albus OAY2 isolated from seaweed.•Fermentation process was optimized with enzyme activity improving to 2.65 U/ml.•Purification resulted high specific activity of 2715 and 1338 U/mg, respectively.•Hydrolysis products of agarose by agarase-a included diverse NAOS: NA2, NA4, NA6.•The products of agarose by agarase-a were different from those of the agarases of GH118 family.

Two extracellular β-agarases were purified from Agarivorans albus OAY2, which was isolated from seaweed collected in Qingdao, Shandong, China. The fermentation process was optimized with enzyme activity improving from 1.06 U/ml to 2.65 U/ml. Agarases were purified by ammonium sulfate fractionation and ion-exchange chromatography. Purification resulted in a 107-fold increase in agarase-a activity and a 52-fold increase in case of agarase-b, with high specific activities of 2715 and 1338 U/mg, respectively. The molecular masses of the agarases were estimated to be 50 kDa and 107 kDa, respectively. The optimum temperatures for the activities of agarase-a and agarase-b were 40 °C and 50 °C, respectively. Agarase-a was stable at 30 °C, while agarase-b was stable at 50 °C. The optimum pH values for agarase-a and agarase-b were both 9.0. Agarase-a was inhibited by Cu2+, Mn2+, Co2+ and EDTA, while agarase-b was inhibited by Cu2+, Mn2+ and Fe2+. The results of 13C NMR and TLC showed that hydrolysis of agarose by agarase-a produced neoagarobiose (NA2), neoagarotetraose (NA4) and neoagarohexaose (NA6). Agarase-b hydrolyzed agarose to yield neoagarobiose (NA2) and neoagarotetraose (NA4). Evaluation of matrix-assisted laser desorption ionization/time-of-flight MS (MALDI-TOF-TOF/MS) and the enzymatic product results indicated that agarase-a and agarase-b might be novel agarases.

Keywords
Purification; Characterization; β-Agarase; Agarivorans; Neoagaro-oligosaccharide
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Purification and characterization of two agarases from Agarivorans albus OAY02
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 49, Issue 5, May 2014, Pages 905–912
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
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