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Mar. Drugs 2014, 12(5), 2731-2747; doi:10.3390/md12052731
Article

Overexpression and Characterization of a Novel Thermostable β-Agarase YM01-3, from Marine Bacterium Catenovulum agarivorans YM01T

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Received: 26 November 2013; in revised form: 21 April 2014 / Accepted: 22 April 2014 / Published: 12 May 2014
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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 β-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 Km of 3.78 mg mL−1 for agarose and a Vmax of 1.14 × 104 U mg−1. YM01-3 hydrolyzed the β-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.
Keywords: Catenovulum agarivorans YM01T; expression; characterization; YM01-3; β-agarase; thermostable Catenovulum agarivorans YM01T; expression; characterization; YM01-3; β-agarase; thermostable
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Cui, F.; Dong, S.; Shi, X.; Zhao, X.; Zhang, X.-H. Overexpression and Characterization of a Novel Thermostable β-Agarase YM01-3, from Marine Bacterium Catenovulum agarivorans YM01T. Mar. Drugs 2014, 12, 2731-2747.

AMA Style

Cui F, Dong S, Shi X, Zhao X, Zhang X-H. Overexpression and Characterization of a Novel Thermostable β-Agarase YM01-3, from Marine Bacterium Catenovulum agarivorans YM01T. Marine Drugs. 2014; 12(5):2731-2747.

Chicago/Turabian Style

Cui, Fangyuan; Dong, Sujie; Shi, Xiaochong; Zhao, Xia; Zhang, Xiao-Hua. 2014. "Overexpression and Characterization of a Novel Thermostable β-Agarase YM01-3, from Marine Bacterium Catenovulum agarivorans YM01T." Mar. Drugs 12, no. 5: 2731-2747.



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