d -氨基酸的生理作用及d -氨基酸氧化酶的生物分析潜力

Vladimir I. Tishkov, Michail D. Shelomov, Anastaiya A. Pometun, Svyatoslav S. Savin, Denis L. Atroshenko
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摘要

d -氨基酸氧化酶(DAAO)在原核生物和真核生物的功能中都起着重要的作用。DAAO在实践中得到越来越多的应用,包括用于测定复杂样品(包括人体组织和液体)中的d -氨基酸。在所有生物体中,DAAO通常有两种类型。第一类是d -天冬氨酸高度特异性的酶,有自己的名字d -天冬氨酸氧化酶(DASPO)。第二种类型的DAAO具有广泛的底物特异性,对一种或另一种d -氨基酸的偏好因来源而异。DAAO与大量底物的活性极大地复杂化了特定d -氨基酸的选择性测定。这个问题通常是通过选择一种酶来解决的,在分析条件下,对样品中存在的其他d -氨基酸具有低活性或没有活性。为了方便选择特定的酶,我们收集并分析了已知daao与最重要的d -氨基酸的催化参数的文献数据。此外,来自甲基营养酵母Ogataea parapolymorpha DL-1的新型重组daao也得到了类似的数据。数据分析表明,在d -氨基酸系中,新的OpaDASPO和OpaDAAO具有最高的催化参数。
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PHYSIOLOGICAL ROLE OF D-AMINO ACIDS AND BIOANALYTICAL POTENTIAL OF D-AMINO ACID OXIDASES
D-amino acid oxidase (DAAO) plays an important role in the functioning of both prokaryotes and eukaryotes. DAAO is increasingly being used in practice, including for the determination of D-amino acids in complex samples, including human tissues and fl uids. There are generally two types of DAAO in all organisms. The fi rst type is an enzyme highly specifi c for D-aspartate and has its own name D-aspartate oxidase (DASPO). DAAO of the second type is characterized by a wide spectrum of substrate specificity, with preference for one or another D-amino acid varying from source to source. The activity of DAAO with a large number of substrates greatly complicates the selective determination of a particular D-amino acid. The problem is often solved by choosing an enzyme that, under the conditions of analysis, has low or no activity with other D-amino acids present in the sample. For the convenience of selecting a particular enzyme, we have collected and analyzed literature data on the catalytic parameters of known DAAOs with the most important D-amino acids. In addition, similar data are presented for novel recombinant DAAOs from the methylotrophic yeast Ogataea parapolymorpha DL-1. Analysis of the data shows that, with the D-amino acid series, the new OpaDASPO and OpaDAAO have the highest catalytic parameters.
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