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Journal of Zhejiang University SCIENCE B
ISSN 1673-1581(Print), 1862-1783(Online), Monthly
2021 Vol.22 No.11 P.941-953
Heme oxygenase-1/carbon monoxide signaling participates in the accumulation of triterpenoids of Ganoderma lucidum
Abstract: Ganoderic triterpenoids (GTs) are the primary bioactive constituents of the Basidiomycotina fungus, Ganoderma lucidum. These compounds exhibit antitumor, anti-hyperlipidemic, and immune-modulatory pharmacological activities. This study focused on GT accumulation in mycelia of G. lucidum mediated by the heme oxygenase-1 (HO-1)/carbon monoxide (CO) signaling. Compared with the control, hemin (10 μmol/L) induced an increase of 60.1% in GT content and 57.1% in HO-1 activity. Moreover, carbon monoxide-releasing molecule-2 (CORM-2), CO donor, increased GT content by 56.0% and HO-1 activity by 18.1%. Zn protoporphyrin IX (ZnPPIX), a specific HO-1 inhibitor, significantly reduced GT content by 26.0% and HO-1 activity by 15.8%, while hemin supplementation reversed these effects. Transcriptome sequencing showed that HO-1/CO could function directly as a regulator involved in promoting GT accumulation by regulating gene expression in the mevalonate pathway, and modulating the reactive oxygen species (ROS) and Ca2+ pathways. The results of this study may help enhance large-scale GT production and support further exploration of GT metabolic networks and relevant signaling cross-talk.
Key words: Ganoderma lucidum; Triterpenoid; Heme oxygenase-1 (HO-1)/carbon monoxide (CO) signaling; Transcriptome sequencing
创新点:HO-1/CO信号的研究多见于模式植物,在微生物中较少。本文主要研究了HO-1/CO信号在灵芝菌三萜积累过程中的作用,并初步证实HO-1/CO信号可参与调控三萜合成途径中关键酶基因的表达,同时可影响灵芝菌活性氧(ROS)和钙离子(Ca2+)信号的转导。
方法:本文首先利用HO-1/CO信号诱导剂及抑制剂处理液体发酵的灵芝菌丝,研究了HO-1/CO信号对三萜含量与HO-1酶活及基因表达的影响;其次,研究了HO-1/CO信号对三萜合成途径中关键酶基因(hmgr、sqs、ls)表达的影响;最后,利用转录组测序技术分析HO-1/CO信号介导下灵芝菌的差异表达基因,重点分析了ROS和Ca2+信号中关键基因的变化特征,并进行逆转录聚合酶链反应(RT-PCR)验证。
结论:与未处理相比,血红素(hemin,10μmol/L)与外源性CO释放分子(CORM-2,10μmol/L)处理下三萜含量分别增加60.1%和56.0%;HO-1酶活分别增加57.1%和18.1%;而HO-1特异性抑制剂锌卟啉(ZnPPIX)则显著降低了三萜含量与HO-1酶活。上述各处理对三萜合成途径中关键酶基因(hmgr、sqs、ls)表达均造成不同程度的影响,初步证实HO-1及CO均参与介导三萜合成,且HO-1主要依靠其催化产物CO实现该调控作用。此外,转录组结果显示,HO-1/CO信号在参与介导三萜合成的过程中,还会影响ROS和Ca2+信号的表达。
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DOI:
10.1631/jzus.B2000818
CLC number:
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On-line Access:
2024-08-27
Received:
2023-10-17
Revision Accepted:
2024-05-08
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