UV-dependent production of 25-hydroxyvitamin D2 in the recombinant yeast cells expressing human CYP2R1.

UV-dependent production of 25-hydroxyvitamin D2 in the recombinant yeast cells expressing human CYP2R1.
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表达人 CYP2R1 的重组酵母细胞中 UV 依赖性 25-羟基维生素 D2 的产生。

DOI:
10.1016/j.bbrc.2013.02.124
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发表时间:
2013
期刊:
Biochem Biophys Res Commun.
影响因子:
--
通讯作者:
et al.
et al.
中科院分区:
--
文献类型:
--
作者:
Yasuda K;et al.

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CYP 2 R1是一种重要的维生素D25-羟化酶。我们已经成功地在酿酒酵母中表达了人CYP 2 R1,以揭示其酶学性质。在这项研究中,我们研究了生产25-羟基化维生素D使用整个重组酵母细胞表达CYP 2 R1。将维生素D3或维生素D2加入到表达CYP 2 R1的酵母细胞在含有葡萄糖和β-环糊精的缓冲液中的细胞悬浮液中时,维生素被转化为它们的25-羟基化产物。接下来,我们用UVB照射细胞悬浮液并在37°C下孵育。令人惊讶的是,25-羟基维生素D2在没有额外的维生素D2的情况下产生。内源性麦角甾醇可能通过紫外线照射和热异构化转化为维生素D2,然后通过CYP 2 R1将所得维生素D2转化为25-羟基维生素D2。这种不需要底物而生产25-羟基维生素D2的新方法可能具有实用价值。
CYP2R1 is known to be a physiologically important vitamin D 25-hydroxylase. We have successfully expressed human CYP2R1 in Saccharomyces cerevisiae to reveal its enzymatic properties. In this study, we examined production of 25-hydroxylated vitamin D using whole recombinant yeast cells that expressed CYP2R1. When vitamin D3or vitamin D2was added to the cell suspension of CYP2R1-expressing yeast cells in a buffer containing glucose and β-cyclodextrin, the vitamins were converted into their 25-hydroxylated products. Next, we irradiated the cell suspension with UVB and incubated at 37°C. Surprisingly, the 25-hydroxy vitamin D2was produced without additional vitamin D2. Endogenous ergosterol was likely converted into vitamin D2by UV irradiation and thermal isomerization, and then the resulting vitamin D2was converted to 25-hydroxyvitamin D2by CYP2R1. This novel method for producing 25-hydroxyvitamin D2without a substrate could be useful for practical purposes.
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