The First Thermophilic α-Oxoamine Synthase Family Enzyme That Has Activities of 2-Amino-3-ketobutyrate CoA Ligase and 7-Keto-8-aminopelargonic Acid Synthase: Cloning and Overexpression of the Gene from an Extreme Thermophile,Thermus thermophilus, and Characterization of Its Gene Product

Detection and Characterization of a Thermophilic Biotin Biosynthetic Enzyme, 7-Keto-8-aminopelargonic Acid Synthase, from Various Thermophiles

“…[26][27][28][29] SytBioF was also found in this study to catalyze the reaction of KBL with acetyl-CoA and glycine as substrates, like the TTHA1582 protein from T. thermophilus, as described previously. 18) In contrast, the other purified BioFs, AqBioF and GkBioF, did not show any activities when we used acyl-CoAs and amino acids other than pimeloyl-CoA and L-alanine as substrates. Thus the gene products annotated as BioF, AqBioF, CltBioF, GkBioF, and TseBioF, have the activity of BioF with narrow substrate specificities, while the other gene products annotated KBL, SytBioF and the TTHA1582 protein, have the activities of both BioF and KBL with broad substrate specificities.…”

“…KAPA synthase activity was measured mainly by a bioassay described previously. 18) The standard assay mixture, consisting of 100 mM potassium phosphate buffer (pH 6.0), 50 mM L-alanine, 0.2 mM pimeloyl-CoA, 0.1 mM PLP, and enzyme solution in a final volume of 50 mL, was incubated at each temperature for 1 h. The amount of KAPA synthesized was determined micro-…”

“…SytbioF is annotated as a KBL gene in the genomic database, and the gene product shares high identity with the T. thermophilus TTHA1582 protein (60% amino acid sequence identity), which has enzyme activity of BioF, as described in a previous report. 18) As shown in Fig. 2, an amino acid sequence alignment of the gene products of the five putative bioFs and E. coli bioF, revealed highly conserved residues, such as the cofactor PLP binding-site (Lys236 in E. coli BioF), the residues co-ordinating the phosphate group of PLP (Ser107, Gly108, and Thr233), and active site residues (Glu175, Ser179, His133, and His207).…”

“…In our previous study, 18) we identified a thermophilic enzyme that has the activity of BioF, from an extreme thermophile, Thermus thermophilus HB8, in searching for homologs of E. coli genes in a genomic database. We found that the TTHA1582 gene of T. thermophilus HB8 codes putative BioF and KBL.…”

“…a In the reactions with acyl-CoA, no activities were detected for the following amino acids: D-alanine, -alanine, L-valine, L-leucine, L-isoleucine, Lglutamic acid, L-aspartic acid, L-arginine, L-threonine, L-cysteine, Lmethionine, L-proline, L-phenylalanine, L-tyrosine, L-tryptophan, and Lhistidine. b Data from previous study 18…”