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首页> 外文期刊>Journal of bacteriology >Distinct Roles for Two CYP226 Family Cytochromes P450 in Abietane Diterpenoid Catabolism by Burkholderia xenovorans LB400
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Distinct Roles for Two CYP226 Family Cytochromes P450 in Abietane Diterpenoid Catabolism by Burkholderia xenovorans LB400

机译：两种CYP226家族细胞色素P450在Abietane二萜类化合物分解代谢中的不同作用（Burkholderia xenovorans LB400）

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The 80-kb dit cluster of Burkholderia xenovorans LB400 encodes the catabolism of abietane diterpenoids. This cluster includes ditQ and ditU, predicted to encode cytochromes P450 (P450s) belonging to the poorly characterized CYP226A subfamily. Using proteomics, we identified 16 dit-encoded proteins that were significantly more abundant in LB400 cells grown on dehydroabietic acid (DhA) or abietic acid (AbA) than in succinate-grown cells. A key difference in the catabolism of DhA and AbA lies in the differential expression of the P450s; DitU was detected only in the AbA-grown cells, whereas DitQ was expressed both during growth on DhA and during growth on AbA. Analyses of insertion mutants showed that ditQ was required for growth on DhA, ditU was required for growth on AbA, and neither gene was required for growth on the central intermediate, 7-oxo-DhA. In cell suspension assays, patterns of substrate removal and metabolite accumulation confirmed the role of DitU in AbA transformation and the role of DitQ in DhA transformation. Spectral assays revealed that DitQ binds both DhA (dissociation constant, 0.98 ± 0.01 μM) and palustric acid. Finally, DitQ transformed DhA to 7-hydroxy-DhA in vitro. These results demonstrate the distinct roles of the P450s DitQ and DitU in the transformation of DhA and AbA, respectively, to 7-oxo-DhA in a convergent degradation pathway.
机译：伯克霍尔德菌xenovorans LB400的80-kb dit 簇编码阿比特尼二萜的分解代谢。该簇包括 ditQ 和 ditU ，预计它们会编码属于特征较弱的CYP226A亚家族的细胞色素P450（P450s）。使用蛋白质组学，我们鉴定了16种 dit 编码的蛋白质，它们在脱氢松香酸（DhA）或松香酸（AbA）上生长的LB400细胞中比在琥珀酸盐中生长的细胞中明显更丰富。 DhA和AbA分解代谢的关键差异在于P450s的差异表达。仅在AbA生长的细胞中检测到DitU，而DitQ在DhA上生长期间和AbA上生长期间均表达。插入突变体的分析表明， ditQ 在DhA上生长是必需的， ditU 在AbA上生长是必需的，而在中间中间体7-氧代-DhA。在细胞悬浮液测定中，底物去除和代谢产物积聚的模式证实了DitU在AbA转化中的作用以及DitQ在DhA转化中的作用。光谱分析显示，DitQ结合DhA（解离常数，0.98±0.01μM）和棕榈酸。最后，DitQ在体外将DhA转化为7-羟基-DhA。这些结果证明了P450 DitQ和DitU在收敛降解途径中分别将DhA和AbA转化为7-氧代-DhA的独特作用。

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