Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Cloning and Expression of Escherichia coli K13 Phytase Gene (appA13) Isolated from Seawater

  • Kim Young-Ok (Biotechnology Research Center, National Fisheries Research and Development Institute) ;
  • Kim Han-Woo (Biotechnology Research Center, National Fisheries Research and Development Institute) ;
  • Lee Jung-Ho (Biotechnology Research Center, National Fisheries Research and Development Institute) ;
  • Kim Kyung-Kil (Biotechnology Research Center, National Fisheries Research and Development Institute) ;
  • Lee Jong-Yun (Biotechnology Research Center, National Fisheries Research and Development Institute)
  • Published : 2003.03.01
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Abstract

A bacterial strain was isolated from seawater to screen for phytase activities. A colony had the highest activity and was identified as an Escherichia coli strain. Using primers derived from E. coli acid phosphatase appA sequence, we cloned a 1,495 bp DNA fragment connected with the pGEM-T vector. It was over-expressed under lac promoter combined with its native promoter in E. coli $DH5\alpha$. The expression of the phytase gene occurred during late exponential growth and the intracellular phytase production was 16.9 units/ml. The yield of recombinant phytate was 412-fold higher than that of wild type E. coli K13.

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References

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