Primary structures of fungal fructosyl amino acid oxidases and their application to the measurement of glycated proteins.
Fructosyl amino acid oxidase (FAOD), which is active toward model compounds of the glycated proteins in blood, N epsilon-fructosyl N sigma-Z-lysine and N-fructosyl valine, was purified to homogeneity from Aspergillus terreus GP1. Though the enzyme did not use glycated proteins directly as its substrate, it used glycated human serum albumin (HSA) when HSA was treated with a protease. Linear relationships between both the concentration and the increase in absorbance and the glycation rate of glycated HSA and the increase in absorbance were observed. cDNAs coding for FAODs were cloned from cDNA libraries of A. terreus GP1 and Penicillium janthinellum AKU 3413. The coding region for both fungal FAODs consisted of 1314 bp encoding 437 amino acids. The sequence of a dinucleotide-binding motif, GXGXXG, was in the deduced N-terminal region and a similar sequence to that the active site of bacterial sarcosine oxidases was found near the C-terminal region of FAOD. The of C-terminal tripeptides SKL and AKL of FAODs from A. terreus and P. janthinellum, respectively, represent typical peroxisomal-targeting signals. Finally, FAOD protein was produced in Escherichia coli transformants in an active form, and at the same level as in the original fungi.
Yoshida
N
N
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.
Sakai
Y
Y
Isogai
A
A
Fukuya
H
H
Yagi
M
M
Tani
Y
Y
Kato
N
N
D000594:Amino Acid Oxidoreductases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology; D001665:Binding Sites; D005800:Genes, Fungal; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010407:Penicillium / Q000201:enzymology*; D015183:Restriction Mapping; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012709:Serum Albumin / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D017127:Glycation End Products, Advanced; D012709:Serum Albumin; D000594:Amino Acid Oxidoreductases; C103012:amadoriase