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Title: Cloning and characterization of farnesyl diphosphate synthase from the rubber-producing mushroom Lactarius chrysorrheus
Authors: Dararat Mekkriengkrai
Tomoki Sando
Kazutake Hirooka
Jitladda Sakdapipanich
Yasuyuki Tanaka
Ei Ichiro Fukusaki
Akio Kobayashi
Mahidol University
The Institute of Science and Technology for Research and Development, Mahidol University
Osaka University
Fukuyama University
Keywords: Biochemistry, Genetics and Molecular Biology;Chemistry;Immunology and Microbiology
Issue Date: 1-Nov-2004
Citation: Bioscience, Biotechnology and Biochemistry. Vol.68, No.11 (2004), 2360-2368
Abstract: Farnesyl diphosphate is involved in rubber biosynthesis as an initiating substrate for both polyprenol and mushroom rubber. So far, we have isolated the cDNA of a farnesyl diphosphate synthase (FPS) for the first time from a rare rubber-producing mushroom, Lactarius chrysorrheus, by the degenerate RT-PCR technique based on sequence information of FPS genes from fungi and yeasts. The open reading frame was clarified to encode a protein of 381 amino acid residues with a calculated molecular weight of 42.9 kDa. The deduced amino acid sequence of L. chrysorrheus FPS showed about 50% identity with those of other fungi and yeasts as well as plants. We expressed the cDNA of L. chrysorrheus FPS in Escherichia coli as a glutathione-S-transferase (GST)-fusion protein. The purified obtained protein showed FPS activity in which geranyl diphosphate (GPP) served as primary substrate, with a 2.4-fold higher kcat/Kmvalue for GPP than for dimethylallyl diphosphate (DMAPP).
ISSN: 09168451
Appears in Collections:Scopus 2001-2005

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