OPTIMIZATION OF HYDROXYLATION OF TYROSINE AND TYROSINE-CONTAINING PEPTIDES BY MUSHROOM TYROSINASE
OPTIMIZATION OF HYDROXYLATION OF TYROSINE AND TYROSINE-CONTAINING PEPTIDES BY MUSHROOM TYROSINASE
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DOI：
10.1016/0167-4838(86)90152-4
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发表时间：
1986-07-25
期刊：
影响因子：
--
通讯作者：
WAITE, JH
中科院分区：
文献类型：
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作者：
MARUMO, K;WAITE, JH
在还原剂存在下，各种多肽和蛋白质中的游离酪氨酸和酪氨酸残基被酪氨酸酶(单酚，L-多巴：氧氧化还原酶，EC 1.14.18.1)转化为多巴和多巴残基。考察了底物与酪氨酸的摩尔比、还原剂和氧气的浓度、pH值、温度和反应时间等条件对酪氨酸-多巴转化效率的影响。羟化反应的最佳条件为：0.1M磷酸盐缓冲液，pH为7，25 mM抗坏血酸，1 mM酪氨酸，50微克/毫升酪氨酸酶，20度。C.在这些条件下，高达70%的游离酪氨酸转化为多巴，几个合成肽中的酪氨酸残基也以与游离酪氨酸一样高的比例羟化为多巴残基。尤其是十肽羟基类似物(Ala-Lys-Pro-Ser-Tyr-Pro-Thr-Tyr-Lys)的制备，可能有助于更好地理解含多巴的贻贝胶蛋白中的粘附性。
Free tyrosine and tyrosine residues in various peptides and proteins are converted into dopa and dopa residues by tyrosinase (monophenol, L-dopa: oxygen oxidoreductase, EC 1.14.18.1) in the presence of reductants. The efficiency of the tyrosine-to-dopa conversion was examined under varied conditions, such as the substrate-to-tyrosine ratio, concentrations of reductant and oxygen in the reaction solution, pH, temperature and reaction time. The highest dopa yields were achieved with the following optimal conditions for hydroxylation: 0.1 M phosphate buffer at pH 7,25 mM ascorbic acid, 1 mM tyrosine, 50 .mu.g/ml tyrosinase and 20.degree. C. Using these conditions, up to 70% of free tyrosine was converted into dopa, and tyrosine residues in several synthetic peptides were also hydroxylated to dopa residues at ratios as high as free tyrosine. The preparation of hydroxylated analogues of the decapeptide (Ala-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-Tyr-Lys), in particular, may contribute to a better understanding of adhesion in the dopa-containing mussel glue protein.