| 朴晟民,李芷汀,李鸿成,等.操纵全局转录调控因子mcrA激活Penicillium sumatraense OUCF143中的次级代谢产物[J].中国海洋药物,2027,(1):-. |
| 操纵全局转录调控因子mcrA激活Penicillium sumatraense OUCF143中的次级代谢产物 |
| Manipulation of the Global Regulator mcrA Upregulates Secondary Metabolite Production in Penicillium sumatraense OUCF143 |
| 投稿时间：2025-05-09 修订日期：2026-01-22 |
| DOI： |
| 中文关键词: 深海冷泉来源真菌 次级代谢产物 遗传转化 全局转录调控因子 抗多重耐药菌活性。 |
| English Keywords:Deep-sea cold seeps derived fungi Secondary metabolites Genetic transformation Global transcriptional regulator Anti-multidrug-resistant bacteria activity. |
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| 中文摘要: |
| 目的：通过敲除全局性转录调控因子mcrA激活深海冷泉来源真菌Penicillium sumatraense OUCF143的沉默基因簇,挖掘新颖的活性天然产物。方法：结合柱层析,液相色谱和波谱学方法对P. sumatraense OUCF143中的次级代谢产物进行分离鉴定。探究P. sumatraense OUCF143的抗生素敏感性,建立遗传转化条件,并敲除全局性转录调控因子mcrA。对突变株中生成的次级代谢产物进行分离鉴定,并测试所有化合物的抗菌活性。结果：从1株深海冷泉来源真菌P. sumatraense OUCF143中鉴定了2个聚酮类化合物,包括 (S)-curvularin(1)和dehydrocurvularin(2)。抗生素敏感性实验表明,P. sumatraense OUCF143在含有100.0 μg/mL的遗传霉素或母液稀释100倍中生菌素的PDA培养基中被抑制。通过PEG介导的原生质体转化方法,在P. sumatraense OUCF143中敲除了全局性转录调控因子mcrA的同源基因g8009,HPLC结果表明生成了多个次级代谢产物。对P. sumatraense OUCF143中新生成的次级代谢产物进行分离鉴定,共获得了3个化合物,分别是daiazein(3)、(+)-2′S-isorhodoptilometrin(4)和emodin(5)。抗菌活性结果表明化合物2对Enterococcus faecium CCARM 5203表现出中等的抗菌活性,其最小抑菌浓度为12.5 μg/mL。结论：从深海冷泉来源真菌P. sumatraense OUCF143中鉴定了2个聚酮类化合物,并通过操纵全局转录调控因子激活了多个次级代谢产物,并对其中的3个化合物进行分离鉴定。为后续通过操纵全局性转录调控策略激活其他菌株中的沉默基因簇提供了借鉴。 |
| English Summary: |
| Objective: To activate silent gene clusters in the deep-sea cold seep fungus Penicillium sumatraense OUCF143 by knocking out the global transcriptional regulator mcrA, thereby facilitating the discovery of novel bioactive natural products. Methods: Secondary metabolites of P. sumatraense OUCF143 were separated and identified using a combination of column chromatography, liquid chromatography, and spectroscopic methods. The antibiotic susceptibility of P. sumatraense OUCF143 was investigated to establish genetic transformation conditions, followed by the knockout of the global transcriptional regulator mcrA. The resulting secondary metabolites were then isolated, identified, and all compounds were tested for their antibacterial activity. Results Two polyketide compounds, (S)-curvularin (1) and dehydrocurvularin (2), were identified from the deep-sea cold seep fungus P. sumatraense OUCF143. Antibiotic susceptibility tests revealed that the growth of P. sumatraense OUCF143 on PDA medium was inhibited in the presence of 100.0 μg/mL geneticin or a 100-fold dilution of nystatin stock solution. The homologous gene g8009 of the global transcriptional regulator mcrA was successfully knocked out in P. sumatraense OUCF143 via PEG-mediated protoplast transformation, and HPLC analysis indicated the production of several new secondary metabolites. Subsequent isolation and identification of these newly produced secondary metabolites yielded three compounds: daiazein (3), (+)-2′S-isorhodoptilometrin (4), and emodin (5). Antibacterial activity assays demonstrated that compound 2 exhibited moderate activity against Enterococcus faecium CCARM 5203, with a minimum inhibitory concentration of 12.5 μg/mL. Conclusion: Two polyketide compounds were identified from the deep-sea cold seep fungus P. sumatraense OUCF143. Furthermore, manipulation of a global transcriptional regulator led to the activation of multiple secondary metabolites, from which three additional compounds were isolated and identified. This study provides a valuable reference for future research aimed at activating silent gene clusters in other microbial strains through the manipulation of global transcriptional regulatory strategies. |
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