Cytochromes P450 (CYPs or P450s) comprise a superfamily of heme‐containing monooxygenases that are involved in a variety of biological processes. CYPs have broad utilities in industry, but most exhibit low thermostability, limiting their use on an industrial scale. Highly thermostable enzymes can be obtained from thermophiles in geothermal areas, including hot springs, offshore oil‐producing wells and volcanoes. Here, we report the identification of a gene encoding for a thermophilic CYP from the Binh Chau hot spring metagenomic database, which was designated as P450‐T2. The deduced amino acid sequence showed the highest identity of 73.15% with CYP203A1 of Rhodopseudomonas palustris, supporting that P450‐T2 is a member of the CYP203A subfamily. Recombinant protein expression yielded 541 nm. The optimal temperature and pH of P450‐T2 were 50 °C and 8.0, respectively. The half‐life of P450‐T2 was 50.2 min at 50 °C, and its melting temperature was 56.80 ± 0.08 °C. It was found to accept electrons from all tested redox partners systems, with BmCPR‐Fdx2 being the most effective partner. Screening for putative substrates revealed binding of phenolic compounds, such as l‐mimosine and emodin, suggesting a potential application of this new thermophilic P450 in the production of the corresponding hydroxylated products.

中文翻译：

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越南平洲温泉 CYP203A 亚科嗜热细胞色素 P450 的表征

细胞色素 P450（CYP 或 P450）包含一个包含血红素的单加氧酶超家族，参与多种生物过程。CYP 在工业中具有广泛的用途，但大多数表现出低热稳定性，限制了它们在工业规模上的使用。高热稳定性酶可以从地热区的嗜热菌中获得，包括温泉、海上采油井和火山。在这里，我们报告了从平洲温泉宏基因组数据库中鉴定出编码嗜热 CYP 的基因，该基因被命名为 P450-T2。推导的氨基酸序列与沼泽红假单胞菌的 CYP203A1 具有 73.15% 的最高同一性，支持 P450-T2 是 CYP203A 亚科的成员。重组蛋白表达产生 541 nm. P450-T2 的最佳温度和 pH 值分别为 50 °C 和 8.0。P450-T2 在 50 °C 下的半衰期为 50.2 min，其熔化温度为 56.80 ± 0.08 °C。发现它接受来自所有测试过的氧化还原伙伴系统的电子，其中 BmCPR-Fdx2 是最有效的伙伴。对推定底物的筛选揭示了酚类化合物的结合，例如l-含羞草碱和大黄素，表明这种新的嗜热 P450 在生产相应的羟基化产物中具有潜在应用。