Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Improvement in the Catalytic Activity of ${\beta}$-Agarase AgaA from Zobellia galactanivorans by Site-Directed Mutagenesis

  • Lee, Seung-Woo (Department of Pharmaceutical Engineering, College of Medical Life Sciences, Silla University) ;
  • Lee, Dong-Geun (Department of Pharmaceutical Engineering, College of Medical Life Sciences, Silla University) ;
  • Jang, Min-Kyung (Department of Pharmaceutical Engineering, College of Medical Life Sciences, Silla University) ;
  • Jeon, Myong-Je (Department of Pharmaceutical Engineering, College of Medical Life Sciences, Silla University) ;
  • Jang, Hye-Ji (Department of Pharmaceutical Engineering, College of Medical Life Sciences, Silla University) ;
  • Lee, Sang-Hyeon (Department of Pharmaceutical Engineering, College of Medical Life Sciences, Silla University)
  • Received : 2011.07.04
  • Accepted : 2011.07.25
  • Published : 2011.11.28
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Abstract

In this study, site-directed mutagenesis was performed on the ${\beta}$-agarase AgaA gene from Zobellia galactanivorans to improve its catalytic activity and thermostability. The activities of three mutant enzymes, S63K, C253I, and S63K-C253I, were 126% (1,757.78 U/mg), 2.4% (33.47 U/mg), and 0.57% (8.01 U/mg), respectively, relative to the wild-type ${\beta}$-agarase AgaA (1,392.61 U/mg) at $40^{\circ}C$. The stability of the mutant S63K enzyme was 125% of the wild-type up to $45^{\circ}C$, where agar is in a sol state. The mutant S63K enzyme produced 166%, 257%, and 220% more neoagarohexaose, and 230%, 427%, and 350% more neoagarotetraose than the wild-type in sol, gel, and nonmelted powder agar, respectively, at $45^{\circ}C$ over 24 h. The mutant S63K enzyme produced 50% more neoagarooligosaccharides from agar than the wild-type ${\beta}$-agarase AgaA from agarose under the same conditions. Thus, mutant S63K ${\beta}$-agarase AgaA may be useful for the production of functional neoagarooligosaccharides.

Keywords

References

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