Transcriptome Analysis and Identification of the Cholesterol Side Chain Cleavage Enzyme BbgCYP11A1 From Bufo bufo gargarizans.
Transcriptome Analysis and Identification of the Cholesterol Side Chain Cleavage Enzyme BbgCYP11A1 From Bufo bufo gargarizans.
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中华大蟾蜍胆固醇侧链裂解酶 BbgCYP11A1 的转录组分析与鉴定
DOI：
10.3389/fgene.2022.828877
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发表时间：
2022
影响因子：
3.7
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中科院分区：
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中华大蟾蜍（Bufo bufo gargarizans Cantor）是我国珍贵的药用动物。蟾蜍二烯内酯是蟾蜍的主要药理活性成分，由B的毒腺产生。中华大蟾蜍蟾蜍二烯内酯是一种具有六元内酯环的强心苷元，具有抗肿瘤、强心、抗炎、抗炎等作用。蟾蜍二烯内酯的生物合成是复杂和不清楚的。本研究探讨了B三种不同组织（皮肤腺、毒腺和肌肉）的转录组。中华大蟾蜍基因组DNA的高通量测序。根据基因的组织特异性表达谱，预测了389个可能参与蟾蜍二烯内酯生物合成途径的候选基因。然后，BbgCYP 11 A1被鉴定为在生产胆固醇的工程酵母中切割该酶的胆固醇侧链。此外，BbgCYP 11 A1的催化活性与各种氧化还原配偶体进行了研究。有趣的是，来自Anememorea asphodeloides的植物NADPH-细胞色素P450还原酶（CPR）显示出比来自B的BbgAdx-2A-BbgAdR显著更高的产量。中华大蟾蜍这些结果为揭示B中蟾蜍二烯内酯生物合成途径提供了一定的分子基础。中华大蟾蜍
Bufo bufo gargarizans Cantor are precious medicinal animals in traditional Chinese medicine (TCM). Bufadienolides as the major pharmacological components are generated from the venomous glands of B. bufo gargarizans. Bufadienolides are one type of cardiac aglycone with a six-member lactone ring and have properties of antitumor, cardiotonic, tonsillitis, and anti-inflammatory. The biosynthesis of bufadienolides is complex and unclear. This study explored the transcriptome of three different tissues (skin glands, venom glands, and muscles) of B. bufo gargarizans by high-throughput sequencing. According to the gene tissue–specific expression profile, 389 candidate genes were predicted possibly participating in the bufadienolides biosynthesis pathway. Then, BbgCYP11A1 was identified as a cholesterol side chain cleaving the enzyme in engineering yeast producing cholesterol. Furthermore, the catalytic activity of BbgCYP11A1 was studied with various redox partners. Interestingly, a plant NADPH-cytochrome P450 reductase (CPR) from Anemarrhena asphodeloides showed notably higher production than BbgAdx-2A-BbgAdR from B. bufo gargarizans. These results will provide certainly molecular research to reveal the bufadienolides biosynthesis pathway in B. bufo gargarizans.