Journal of Microbiology and Biotechnology

  • 제20권6호
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  • Pages.995-1000
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  • 2010
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Expression and Characterization of a Novel Deoxyribose 5-Phosphate Aldolase from Paenibacillus sp. EA001

  • Kim, Yong-Mo (Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institutee, Korea Research Institute of Bioscience and Biotechnology) ;
  • Choi, Nack-Shick (Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institutee, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Yong-Ook (Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institutee, Korea Research Institute of Bioscience and Biotechnology) ;
  • Son, Dong-Ho (Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institutee, Korea Research Institute of Bioscience and Biotechnology) ;
  • Chang, Young-Hyo (Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Song, Jae-Jun (Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institutee, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Joong-Su (Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institutee, Korea Research Institute of Bioscience and Biotechnology)
  • 투고 : 2009.12.01
  • 심사 : 2010.02.03
  • 발행 : 2010.06.28
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초록

A novel deoC gene was identified from Paenibacillus sp. EA001 isolated from soil. The gene had an open reading frame (ORF) of 663 base pairs encoding a protein of 220 amino acids with a molecular mass of 24.5 kDa. The amino acid sequence was 79% identical to that of deoxyribose 5-phosphate aldolase (DERA) from Geobacillus sp. Y412MC10. The deoC gene encoding DERA was cloned into an expression vector and the protein was expressed in Escherichia coli. The recombinant DERA was purified using Ni-NTA affinity chromatography and then characterized. The optimum temperature and pH of the enzyme were $50^{\circ}C$ and 6.0, respectively. The specific activity for the substrate deoxyribose 5-phosphate (DR5P) was $62\;{\mu}mol/min/mg$. The $K_m$ value for DR5P was determined to be 145 mM with the $k_{cat}$ value of $3.2{\times}10^2/s$ from Lineweaver-Burk plots. The EA001 DERA showed stability toward a high concentration of acetaldehyde (100 mM).

키워드

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  3. Characterization and application of a newly synthesized 2-deoxyribose-5-phosphate aldolase vol.40, pp.1, 2010, https://doi.org/10.1007/s10295-012-1213-y
  4. Biocatalytic Methods for CC Bond Formation vol.5, pp.6, 2010, https://doi.org/10.1002/cctc.201200709
  5. Mechanism-based inhibition of an aldolase at high concentrations of its natural substrate acetaldehyde: structural insights and protective strategies vol.7, pp.7, 2010, https://doi.org/10.1039/c5sc04574f
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  7. 2-Deoxy- D -ribose-5-phosphate aldolase (DERA): applications and modifications vol.102, pp.23, 2018, https://doi.org/10.1007/s00253-018-9392-8