Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Comparison of Properties of the C-Terminus Truncated Agarase of Pseudomonas sp. W7

  • Yoon, Soo-Cheol (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Lee, Jong-Hee (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Ahn, Sun-Hee (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Lee, Eun-Mi (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Park, Eun-Mi (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kong, In-Soo (Department of Biotechnology and Bioengineering, Pukyong National University)
  • Published : 2003.10.01
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Abstract

Three plasmids derived from the ${\beta}-agarase$ gene (PjaA) of Pseudomonas sp. W7 were expressed in Escherichia coli AD494(DE3) pLysS with lactose as an inducer. These products corresponded to the complete (PjaA) and the two C-terminal truncated (PjaAI and PjaAII) forms of ${\beta}-agarase$. The PjaAI and the PjaAII were originated from exonuclease L treatment from PjaA by deleting 127 and 182 amino acid residues-encoded nucleic acids at 3' region, respectively. The molecular weights of the purified proteins were 71 kDa, 58 kDa, and 50 kDa on SDS-PAGE, respectively. The $K_m$ value of PjaAI was lower than that of the PjaA, and the catalytic efficiency ($k_{cat}/K_m$) of PjaAI was increased to 5 times. The enzyme of PjaAI retained more than 90% activity at $50^{\circ}C$. In contrast to the PjaAI, the remaining activity of the PjaA was only 20% at the same temperature.

Keywords

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