Combinatorial alanine substitution enables rapid optimization of cytochrome P450BM3 for selective hydroxylation of large substrates.

Combinatorial alanine substitution enables rapid optimization of cytochrome P450BM3 for selective hydroxylation of large substrates.
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DOI:
10.1002/cbic.201000565
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发表时间:
2010-12-10
期刊:
影响因子:
3.2
通讯作者:
Arnold, Frances H.
Arnold, Frances H.
中科院分区:
生物学3区
文献类型:
--
作者:
Lewis, Jared C.;Mantovani, Simone M.;Fu, Yu;Snow, Christopher D.;Komor, Russell S.;Wong, Chi-Huey;Arnold, Frances H.

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使用热稳定细胞色素P450 BM 3(BM 3)变体中的活性位点残基的组合丙氨酸取代来产生对大底物具有活性的BM 3变体。甲氧基甲基化的单糖、生物碱和甾体的选择性羟基化因此成为可能。这种方法通常可用于改善在较大底物上仅显示有限活性的酶的活性。
Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450BM3 (BM3) variant was used to generate BM3 variants with activity on large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible. This approach could be generally useful for improving the activity of enzymes that show only limited activity on larger substrates.
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