Dataset Information

Overexpression and characterization of a novel thermostable ?-agarase YM01-3, from marine bacterium Catenovulum agarivorans YM01(T).

ABSTRACT: Genome sequencing of Catenovulum agarivorans YM01T reveals 15 open-reading frames (ORFs) encoding various agarases. In this study, extracellular proteins of YM01T were precipitated by ammonium sulfate and separated by one-dimensional gel electrophoresis. The results of in-gel agarase activity assay and mass spectrometry analysis revealed that the protein, YM01-3, was an agarase with the most evident agarolytic activity. Agarase YM01-3, encoded by the YM01-3 gene, consisted of 420 amino acids with a calculated molecular mass of 46.9 kDa and contained a glycoside hydrolase family 16 ?-agarase module followed by a RICIN superfamily in the C-terminal region. The YM01-3 gene was cloned and expressed in Escherichia coli. The recombinant agarase, YM01-3, showed optimum activity at pH 6.0 and 60 °C and had a K(m) of 3.78 mg mL?¹ for agarose and a Vmax of 1.14 × 10? U mg?¹. YM01-3 hydrolyzed the ?-1,4-glycosidic linkages of agarose, yielding neoagarotetraose and neoagarohexaose as the main products. Notably, YM01-3 was stable below 50 °C and retained 13% activity after incubation at 80 °C for 1 h, characteristics much different from other agarases. The present study highlights a thermostable agarase with great potential application value in industrial production.

SUBMITTER: Cui F

PROVIDER: S-EPMC4052312 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

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Overexpression and characterization of a novel thermostable β-agarase YM01-3, from marine bacterium Catenovulum agarivorans YM01(T).

Cui Fangyuan F Dong Sujie S Shi Xiaochong X Zhao Xia X Zhang Xiao-Hua XH

Marine drugs 20140512 5

Genome sequencing of Catenovulum agarivorans YM01T reveals 15 open-reading frames (ORFs) encoding various agarases. In this study, extracellular proteins of YM01T were precipitated by ammonium sulfate and separated by one-dimensional gel electrophoresis. The results of in-gel agarase activity assay and mass spectrometry analysis revealed that the protein, YM01-3, was an agarase with the most evident agarolytic activity. Agarase YM01-3, encoded by the YM01-3 gene, consisted of 420 amino acids wit ...[more]

PMID: 24824021

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Dataset Information

Overexpression and characterization of a novel thermostable ?-agarase YM01-3, from marine bacterium Catenovulum agarivorans YM01(T).

Publications

Overexpression and characterization of a novel thermostable β-agarase YM01-3, from marine bacterium Catenovulum agarivorans YM01(T).

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