Origin of D-amino acids detected in the acid hydrolysates of purified Escherichia coli ß-galactosidase.
Origin of D-amino acids detected in the acid hydrolysates of purified Escherichia coli ß-galactosidase.
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纯化大肠杆菌酸水解产物中检测到的 D-氨基酸来源
DOI：
10.1016/j.jpba.2015.04.022
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
H.
中科院分区：
文献类型：
--
作者：
Miyamoto;T.;Sekine;M.;Ogawa;T.;Hidaka;M.;Homma;H.;Masaki;H.
在以前的报道中，我们在纯化的重组β-半乳糖苷酶的酸水解产物中检测到了DD-氨基酸。在这里，我们采用了氘-氢交换方法来区分从那些水解孵育过程中产生的先天氨基酸。β-半乳糖苷酶在DCl/D2 O中水解后，氨基酸经NBD-F衍生化，在反相柱上分离，然后用配备手性柱的液相色谱-串联质谱法进行分离。我们的研究结果表明，在蛋白质中的先天氨基酸残基的情况下，并建议蛋白质经历异构化在水解孵育的非常早期的阶段。
In previous report, we detectedd-amino acids in the acid hydrolysates of purified recombinant β-galactosidase. Here, we employed a deuterium-hydrogen exchange method to discriminate innated-amino acids from those generated during hydrolytic incubation. After hydrolysis of β-galactosidase in DCl/D2O, amino acids were derivatized with NBD-F and separated on a reverse-phase column, followed by liquid chromatography-tandem mass spectrometry equipped with a chiral column. Our results show an absence of innated-amino acid residues in the protein and suggest that the protein undergoes isomerization during a very early stage of hydrolytic incubation.