Identification and functional analysis of carbonyl reductases related to tetrahydrobiopterin synthesis in the silkworm, Bombyx mori
Identification and functional analysis of carbonyl reductases related to tetrahydrobiopterin synthesis in the silkworm, Bombyx mori
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家蚕四氢生物蝶呤合成相关羰基还原酶的鉴定及功能分析
DOI：
10.1111/imb.12768
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发表时间：
2022-03-04
影响因子：
2.6
通讯作者：
Meng, Yan
中科院分区：
文献类型：
--
作者：
Liang, Dan;Shu, Rui;Meng, Yan
短链脱氨酶/还原酶（SDRs）超家族在生物合成和信号传导途径中至关重要，其中羰基还原酶（CBRs）亚家族在四氢生物蝶呤（BH 4）的生物合成中起重要作用。BH 4是动物必需的辅酶，其缺乏可导致神经系统疾病。家蚕CBRs参与BH_4合成的研究报道较少。在此，我们在B中鉴定了67个SDR基因。家蚕（BmSDR）的全基因组研究。在生物信息学分析和KEGG验证的基础上，对4个可能与BH 4合成相关的BmCBRs进行了进一步的鉴定和功能分析。结果表明，这4个基因在BH 4合成缺陷的lem突变体ah 09的头部和生殖腺中均有高表达。胞内干扰BmCBR和B的主要催化酶sepiapterin还原酶后酶活性、BH 4含量及相关基因表达水平。mori（BmSpr）在BH 4从头途径中的作用表明BmCBR 2在补救途径中起作用。BmCBR 3和BmCBR 4通过旁路途径调节BH 4的合成。在家蚕BH 4合成的4条途径中，从头途径占主导地位，其次是替代途径和补救途径。根据BmSpr干扰后BmCBR 3的过表达，BH 4含量没有显著变化。推测BmCBR 3位于BmSpr的上游。这些结果为深入探讨BmSDR在B中的作用提供了理论基础。也为其他动物相关疾病的研究提供了线索。
The superfamily of short-chain dehydrogenases/reductases (SDRs) is crucial in biosynthetic and signalling pathways, in which the carbonyl reductases (CBRs) subfamily is important in the biosynthesis of tetrahydrobiopterin (BH4). BH4 is an essential coenzyme for animals, and its deficiency can lead to neurological diseases. There are few reports on CBRs involved in BH4 synthesis of silkworms, Bombyx mori. Here, we identified 67 SDR genes in B. mori (BmSDR) through whole genome survey for the first time. Based on bioinformatics analyses and KEGG verification, four BmCBRs that may be related to BH4 synthesis were further characterized and functionally analysed. The results showed these four genes were high expressed in the head and gonads of ah09 (a lem mutant with defective BH4 synthesis). Enzyme activity, BH4 content and the related gene expression levels after intracellular interference with BmCBR and the main catalytic enzymes sepiapterin reductase of B. mori (BmSpr) in the de novo pathway of BH4 showed BmCBR2 plays a role in the salvage pathway. BmCBR3 and BmCBR4 regulate BH4 synthesis through the alternative pathway. Among the four pathways of silkworm BH4 synthesis, the de novo pathway occupies the dominant position, followed by the alternative pathway and salvage pathway. According to the overexpression of BmCBR3 after interference with BmSpr, the BH4 content did not change significantly. It is speculated that BmCBR3 is located upstream of BmSpr. These results provide a theoretical basis for in-depth exploration of the role of BmSDR in B. mori and also provide clues for the research of other animal-related diseases.