Overexpression and molecular characterization of Aga50D from Saccharophagus degradans 2-40: An exo-type β-agarase producing neoagarobiose

Hee Taek Kim, Saeyoung Lee, Dongho Lee, Hyun Soo Kim, Won Gi Bang, Kyoung Heon Kim, In Geol Choi

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

β-Agarases are mostly categorized into three glycoside hydrolase (GH) families 16, 50, and 86. Recent genomic analysis of Saccharophagus degradans 2-40 revealed the presence of five agarase genes belonging to these GH families. Among the five agarases, Aga50D (a member of GH50) had neither been functionally characterized nor overexpressed. In this report, we present soluble overexpression and molecular characterization of Aga50D. Aga50D was expressed in an active form resulting in a single major product from agarose without intermediates. While known GH50 agarases have both endo-lytic and exo-lytic activities, which produce neoagarobiose as a final product through the intermediate, neoagaro-oligosaccharides, identification and analysis of the reaction product by mass spectrometry and 13C NMR showed that Aga50D had unique exo-lytic activity and was able to produce neoagarobiose directly from agarose. The optimum pH and temperature for the activity were 7.0 and 30°C, respectively. The K m and V max for agarose were 41.9 mg/ml (4.2 mM) and 17.9 U/mg, respectively.

Original languageEnglish
Pages (from-to)227-234
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume86
Issue number1
DOIs
Publication statusPublished - 2010 Mar

Keywords

  • Exo-lytic activity
  • GH50
  • Neoagarobiose
  • Saccharification
  • β-Agarase

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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