25023-94-3Relevant articles and documents
Overexpression and characterization of a novel thermostable β-agarase YM01-3, from marine bacterium Catenovulum agarivorans YM01T
Cui, Fangyuan,Dong, Sujie,Shi, Xiaochong,Zhao, Xia,Zhang, Xiao-Hua
, p. 2731 - 2747 (2014/06/10)
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.
Substrate recognition and hydrolysis by a family 50 exo-β-agarase, aga50D, from the marine bacterium Saccharophagus degradans
Pluvinage, Benjamin,Hehemann, Jan-Hendrik,Boraston, Alisdair B.
, p. 28078 - 28088 (2013/10/08)
Background: The catalytic mechanism and substrate recognition required for exo-β-agarase activity are unclear. Results: Structural analysis of a family 50 glycoside hydrolase details substrate recognition and supports a retaining mechanism. Conclusion: The active site architecture dictates exo-activity, whereas substrate distortion aids glycosidic bond hydrolysis. Significance: This structural work offers the first snapshots of Michaelis complexes formed during hydrolysis of the β-linkages in agarose.