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KMID : 0613820120220081024
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Journal of Life Science
2012 Volume.22 No. 8 p.1024 ~ p.1033
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Cloning of Geranylgeranyl Pyrophosphate Synthase (CrtE) Gene from Kocuria gwangalliensis and Its Functional Co-expression in Escherichia coli
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Seo Yong-Bae
Kim Gun-Do
Lee Jae-Hyung
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Abstract
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A gene encoding a novel geranylgeranyl pyrophosphate (GGPP) synthase from Kocuria gwangalliensis has been cloned and expressed in Escherichia coli. The deduced amino acid sequence showed 59.6% identity with a putative GGPP synthase (CrtE) from K. rhizophila. An expression plasmid containing the crtE gene was constructed, and E. coli cells containing this plasmid produced a recombinant protein with a theoretical molecular mass of 41 kDa, corresponding to the molecular weight of GGPP synthase. Due to the lack of crtE, crtB, and crtI in E. coli, the biosynthesis of lycopene was only obtained when the plasmid pCcrtE was co-transformed into E. coli expressing the pRScrtBI-carrying carotenoid biosynthesis crtB and crtI genes, which were sub-cloned from Paracoccus haeundaensis. The biochemical studies on the expressed proteins were performed via HPLC. The results obtained from this study will provide a wider base of knowledge regarding the primary structure of CrtE cloned from K. gwangalliensis at the molecular level.
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KEYWORD
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Kocuria gwangalliensis, carotenoid, geranylgeranyl pyrophosphate (GGPP), lycopene, geranylgeranyl pyrophosphate synthase (CrtE)
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