Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Overexpression and Characterization of Lycopene Cyclase (CrtY) from Marine Bacterium Paracoccus haeundaensis

  • Jeong, Tae Hyug (Department of Microbiology, Pukyong National University) ;
  • Ji, Keunho (Department of Microbiology, Pukyong National University) ;
  • Kim, Young Tae (Department of Microbiology, Pukyong National University)
  • Received : 2012.08.27
  • Accepted : 2012.10.02
  • Published : 2013.02.28
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Abstract

Lycopene cyclase converts lycopene to ${\beta}$-carotene by catalyzing the formation of two beta-rings at each end of the linear carotene structure. This reaction takes place as a two-step reaction in which both sides of of the lycopene molecule are cyclized into ${\beta}$-carotene rings via the monocyclic ${\gamma}$-carotene as an intermediate. The crtY gene coding for lycopene cyclase from Paracoccus haeundaensis consists of 1,158 base pairs encoding 386 amino acids residues. An expression plasmid containing the crtY gene (pET44a-CrtY) was constructed and expressed in Escherichia coli, and produced a recombinant protein of approximately 43 kDa, corresponding to the molecular mass of lycopene cyclase. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to lycopene cyclase. We also determined the lycopene substrate specificity and NADPH cofactor requirements of the purified protein. The $K_m$ values for lycopene and NADPH were 3.5 ${\mu}M$ and 2 mM, respectively. The results obtained from this study will provide a wider base of knowledge on the enzyme characterization of lycopene cyclase at the molecular level.

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References

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