Bacillus subtilis ypgA gene is fni, a nonessential gene encoding type 2 isopentenyl diphosphate isomerase.
We previously identified the fni gene of Streptomyces sp. strain CL190 as type 2 isopentenyl diphosphate (IPP) isomerase, which needs both FMN and NADPH for enzyme activity. An fni gene homolog, ypgA, was detected in the database of the Bacillus subtilis genome. However, the ypgA product was about 140 amino acids shorter in the N-terminal than the Streptomyces fni gene product. A database search found three new putative start codons in 129, 225, and 411 bases upstream of the original start codon of the ypgA gene. The longest gene product, which was named ypgA3, showed the most significant homology to the Streptomyces fni gene product. The ypgA3 gene was expressed with an N-terminal His-tag in Escherichia coli and the purified soluble protein was characterized in detail. The ypgA3 protein converted IPP to its isomer dimethylallyl diphosphate in the presence of both FMN and NADPH. The enzyme also catalyzed the reverse reaction in the presence of both the cofactors. Disruption of the ypgA3 gene was not lethal to B. subtilis. These results indicate that Bacillus ypgA3 gene is fni, a nonessential gene encoding type 2 IPP isomerase.
Takagi
Motoki
M
Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Kaneda
Kazuhide
K
Shimizu
Tomohiro
T
Hayakawa
Yoichi
Y
Seto
Haruo
H
Kuzuyama
Tomohisa
T
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D019748:Carbon-Carbon Double Bond Isomerases / Q000235:genetics* / Q000378:metabolism; D002455:Cell Division / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005486:Flavin Mononucleotide; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D045782:Hemiterpenes; D008969:Molecular Sequence Data; D009154:Mutation; D009249:NADP / Q000378:metabolism; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D045782:Hemiterpenes; D011994:Recombinant Proteins; D009249:NADP; D005486:Flavin Mononucleotide; D019748:Carbon-Carbon Double Bond Isomerases; C021453:isopentenyldiphosphate delta-isomerase