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Cloning, purification and biochemical characterization of beta agarase from the marine bacterium Pseudoalteromonas sp. AG4

  • Chulhong Oh
  • Chamilani Nikapitiya
  • Youngdeuk Lee
  • Ilson Whang
  • Se-Jae Kim
  • Do-Hyung Kang
  • Jehee Lee
Original Paper

Abstract

A gene (agrP) encoding a β-agarase from Pseudoalteromonas sp. AG4 was cloned and expressed in Escherichia coli. The agrP primary structure consists of an 870-bp open reading frame (ORF) encoding 290 amino acids (aa). The predicted molecular mass and isoelectric point were determined at 33 kDa and 5.9, respectively. The signal peptide was predicted to be 21 aa. The deduced aa sequence showed 98.6% identity to β-agarase from Pseudoalteromonas atlantica. The recombinant protein was purified as a fusion protein and biochemically characterized. The purified β-agarase (AgaP) had specific activity of 204.4 and 207.5 units/mg towards agar and agarose, respectively. The enzyme showed maximum activity at 55°C and pH 5.5. It was stable at pH 4.5 to 8.0 and below 55°C for 1 h. The enzyme produced neoagarohexaose and neoagarotetraose from agar and in addition to that neoagarobiose from the agarose. The neoagarooligosaccharides were biologically active. Hence, AgaP is a useful enzyme source for use by cosmetic and pharmaceutical industries.

Keywords

Beta agarase GHF-16 Neoagarooligosaccharides Pseudoalteromonas spp. 

Notes

Acknowledgment

This research was financially supported by the Ministry of Education, Science Technology (MEST) and Korea Institute for Advancement of Technology (KIAT) through the Human Resource Training Project for Regional Innovation, and by a research grant (PP00740) from Korea Ocean Research & Development Institute.

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Copyright information

© Society for Industrial Microbiology 2010

Authors and Affiliations

  • Chulhong Oh
    • 1
    • 4
  • Chamilani Nikapitiya
    • 1
  • Youngdeuk Lee
    • 1
  • Ilson Whang
    • 2
  • Se-Jae Kim
    • 2
  • Do-Hyung Kang
    • 4
  • Jehee Lee
    • 1
    • 3
  1. 1.Department of Marine Life ScienceJeju National UniversityJejuRepublic of Korea
  2. 2.Department of Life ScienceJeju National UniversityJejuRepublic of Korea
  3. 3.Marine and Environmental InstituteJeju National UniversityJejuRepublic of Korea
  4. 4.Korea Ocean Research and Development InstituteAnsanRepublic of Korea

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