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

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Journal of Industrial Microbiology & Biotechnology

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.

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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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Authors and Affiliations

  1. Department of Marine Life Science, Jeju National University, 66 Jejudaehakno, Ara-Dong, Jeju, 690-756, Republic of Korea

    Chulhong Oh, Chamilani Nikapitiya, Youngdeuk Lee & Jehee Lee

  2. Department of Life Science, Jeju National University, 66 Jejudaehakno, Ara-Dong, Jeju, 690-756, Republic of Korea

    Ilson Whang & Se-Jae Kim

  3. Marine and Environmental Institute, Jeju National University, Jeju, 690-814, Republic of Korea

    Jehee Lee

  4. Korea Ocean Research and Development Institute, Ansan, 426-744, Republic of Korea

    Chulhong Oh & Do-Hyung Kang

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  1. Chulhong Oh
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  2. Chamilani Nikapitiya
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Correspondence to Jehee Lee.

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Oh, C., Nikapitiya, C., Lee, Y. et al. Cloning, purification and biochemical characterization of beta agarase from the marine bacterium Pseudoalteromonas sp. AG4. J Ind Microbiol Biotechnol 37, 483–494 (2010). https://doi.org/10.1007/s10295-010-0694-9

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  • DOI: https://doi.org/10.1007/s10295-010-0694-9

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