Gene cloning and overexpression of a geranylgeranyl diphosphate synthase of an extremely thermophilic bacterium, Thermus thermophilus.
A geranylgeranyl diphosphate (GGPP) synthase gene of an extremely thermophilic bacterium, Thermus thermophilus, was cloned and sequenced. T. thermophilus GGPP synthase, overexpressed in Escherichia coli cells as a glutathione S-transferase fusion protein, was purified and characterized. The fusion protein, retaining thermostability, formed a homodimer, and showed higher specific activity than did a partially purified thermostable enzyme previously reported. Optimal reaction conditions and kinetic parameters were also examined. The deduced amino acid sequence indicated that T. thermophilus GGPP synthase was excluded from the group of bacterial type GGPP synthases and lacked the insertion amino acid residues in the first aspartate-rich motif as do archaeal and eukaryotic short-chain prenyltransferases.
Ohto
C
C
Bio Research Laboratory, Toyota Motor Corporation, Japan.
Ishida
C
C
Koike-Takeshita
A
A
Yokoyama
K
K
Muramatsu
M
M
Nishino
T
T
Obata
S
S
D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D015139:Blotting, Southern; D002846:Chromatography, Affinity; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D051231:Farnesyltranstransferase; D015964:Gene Expression Regulation, Bacterial; D005982:Glutathione Transferase / Q000737:chemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016963:Thermus thermophilus / Q000201:enzymology / Q000235:genetics*
D015342:DNA Probes; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D019883:Alkyl and Aryl Transferases; D005982:Glutathione Transferase; D051231:Farnesyltranstransferase