Akademik Veri Yönetim Sistemi
Biotechnology and Biotechnological Equipment, cilt.20, sa.3, ss.76-82, 2006 (Scopus)
Geranylgeranyl pyrophosphate synthase (GGPPS) is a key enzyme in carotenoid biosynthetic pathway, catalyzing the synthesis of its C20 precursor. The gene encoding GGPPS from bell pepper (Capsicum annuum) has been cloned in Schizosaccharomyces pombe and its heterologous expression analyzed in this model organism for eukaryotes. We aim to partially direct the ergosterol pathway to carotenoid biosynthesis in this yeast. This yeast is not able to synthesize carotenoids naturally, but it produces farnesyl pyrophosphate (FPP), an intermediate metabolite for both ergosterol and geranylgeranyl pyrophosphate (GGPP) biosynthesis. S. pombe transformants carrying the pepper GGPPS cDNA were named KTG. In one of the transformant (KTG1), heterologous expression of GGPPS gene has been analyzed. The GGPPS gene was shown by dot and Northern hybridization to be successfully transcribed. When water soluble proteins isolated from the transformant cells have been analyzed on SDS-denaturating gels, a ~37 kDaprotein band has been detected which was not found in the host. In order to understand whether this heterologous protein in KTG1 transformant is an active GGPPS or not, changes in ergosterol level have been analyzed by HPLC. 50–60% decrease in ergosterol content was found in the KTG1 transformant when compared to the host. These results suggest that production of GGPP derivatives can now be attempted, for instance carotenoids, through heterologous expression of carotenoid biosynthetic genes in a non-carotenogenic yeast, S. pombe. © 2006 Taylor and Francis Group, LLC.