Sakaue Ryoichi, Hiruma Minoru, Kajiyama Naoki, Koyama Yasuji
Research and Development Division, Kikkoman Corporation, Noda, Chiba, Japan.
Biosci Biotechnol Biochem. 2002 Jun;66(6):1256-61. doi: 10.1271/bbb.66.1256.
The gene encoding the fructosyl-amino acid oxidase (fructosyl-alpha-L-amino acid: oxygen oxidoreductase (defructosylating); EC 1.5.3) of Corynebacterium sp. 2-4-1 was cloned and expressed in Escherichia coli. The gene consists of 1,116 nucleotides and encodes a protein of 372 amino acids with a predicted molecular mass of 39,042. The open reading frame was confirmed as the gene of the fructosyl-amino acid oxidase by comparison with the N-terminal amino acid sequence of the purified fructosyl-amino acid oxidase from Corynebacterium sp. 2-4-1. The sequence of the AMP-binding motif, GXGXXG, was found in the deduced N-terminal region. The amino acid sequence of the fructosyl-amino acid oxidase showed no similarity to that of fungal fructosyl-amino acid oxidases. In addition, substrate specificities of this fructosyl-amino acid oxidase were different from those of other fructosyl-amino acid oxidases. The fructosyl-amino acid oxidase of Corynebacterium sp. 2-4-1 is an enzyme that has unique substrate specificity and primary structure in comparison with fungal fructosyl-amino acid oxidases.
克隆了棒状杆菌属2-4-1编码果糖基氨基酸氧化酶（果糖基-α-L-氨基酸：氧氧化还原酶（去果糖基化）；EC 1.5.3）的基因，并在大肠杆菌中进行表达。该基因由1116个核苷酸组成，编码一个含有372个氨基酸的蛋白质，预测分子量为39042。通过与从棒状杆菌属2-4-1纯化的果糖基氨基酸氧化酶的N端氨基酸序列进行比较，确认该开放阅读框为果糖基氨基酸氧化酶的基因。在推导的N端区域发现了AMP结合基序GXGXXG的序列。果糖基氨基酸氧化酶的氨基酸序列与真菌果糖基氨基酸氧化酶的序列没有相似性。此外，这种果糖基氨基酸氧化酶的底物特异性与其他果糖基氨基酸氧化酶不同。与真菌果糖基氨基酸氧化酶相比，棒状杆菌属2-4-1的果糖基氨基酸氧化酶是一种具有独特底物特异性和一级结构的酶。
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