Studies on the Chemical Epigenetic Modification of Fungus Samsoniella Hepiali CDB9-31
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摘要:
目的 采用薄层色谱法（thin-layer chromatography，TLC）和高效液相色谱法（high-performance liquid chromatography，HPLC）分析组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸（suberoylanilide hydroxamic acid，SAHA）对虫生真菌Samsoniella hepiali CDB9-31次生代谢产物的影响。 方法 利用硅胶柱层析、葡聚糖凝胶柱、反相柱层析等方法对添加了表观遗传试剂的Samsoniella hepiali CDB9-31发酵产物进行分离、纯化，并利用现代波谱分析方法对化合物进行结构鉴定，采用滤纸片扩散法测定所得单体化合物的抗菌活性。 结果 对其发酵提取物进行TLC和HPLC分析，发现SAHA可以诱导该菌株产生丰富的次生代谢产物，并分离得到11个单体化合物，分别鉴定为：N'-phenyloctanediamide（1）、5-Phenylcarbamoyl-pentanoic（2）、ergosterol（3）、5，8-Epidioxy-5α，8α-ergosta-6，22E-diene-3β-ol（4）、1-monolinolein（5）、（4E，8E）-2-N-（2-Hydroxypalmitoyl）-1-O-（β-D-glucopyranosyl）-9-methyl-4，8-sphingadienine（6）、Ergosterol peroxide 3-O-β-D-glucopyranoside（7）、（22E，24R）-7，22-diene-3β，5α，6β-ergostatriol（8）、（2S，2'R，3R，4E，8E）-N-2'-Hydroxyhexadecanoyl-2-amion-9-methyl-4，8-octadecadiene-1，3-diol（9）、Adenosine（10）、D-Glulopyranose（11）。化合物1和2是SAHA的衍生物，推测是CDB9-31特殊的代谢环境引起了SAHA的生物转化，除了化合物3，其余化合物均为首次从该属中分离得到。抗菌活性结果显示，其中有6个化合物至少对一种病原菌表现出了不同程度的抑制作用。 结论 本研究丰富了虫生真菌Samsoniella hepiali次生代谢产物化学多样性。 -
关键词:
- 表观遗传修饰 /
- 虫生真菌 /
- Samsoniella hepiali /
- 次生代谢产物 /
- 抗菌活性
Abstract:Objective To analyze the effects of the histone deacetylase inhibitor suberoylanilide hydroxamic acid （SAHA） on the secondary metabolites of the entomopathogenic fungus Samsoniella hepiali CDB9-31 using thin-layer chromatography （TLC） and high-performance liquid chromatography （HPLC）. Methods The fermentation products of Samsoniella hepiali CDB9-31 treated with epigenetic modifiers were separated and purified using methods such as silica gel column chromatography, Sephadex column chromatography and reversed-phase column chromatography. The structures of the compounds were elucidated using modern spectroscopic analysis methods. The antimicrobial activity of the obtained monomeric compounds was determined using the filter paper disc diffusion method. Results The TLC and HPLC analyses of its fermentation extracts revealed that SAHA could induce the strain to produce more diverse array of secondary metabolites, and 11 monomeric compounds were isolated and identified as follows: N'-phenyloctanediamide （1）, 5-Phenylcarbamoyl-pentanoic （2）, ergosterol （3）, 5, 8-Epidioxy-5α, 8α-ergosta-6, 22E-diene-3β-ol （4）, 1-monolinolein （5）, （4E, 8E）-2-N-（2-Hydroxypalmitoyl）-1-O-（β-D-glucopyranosyl）-9-methyl-4, 8-sphingadienine （6）, Ergosterol peroxide 3-O-β-D-glucopyranoside （7）, （22E, 24R）-7, 22-diene-3β, 5α, 6β-ergostatriol （8）, （2S, 2'R, 3R, 4E, 8E）-N-2'-Hydroxyhexadecanoyl-2-amion-9-methyl-4, 8-octadecadiene-1, 3-diol （9）, Adenosine （10）, D-Glulopyranose （11）. Compounds 1 and 2 were derivatives of SAHA, and it was speculated that the special metabolic environment of CDB9-31 caused the biotransformation of SAHA. Except for compound 3, all other compounds were isolated from this genus for the first time. The antibacterial activity results showed that six of these compounds exhibited varying degrees of inhibitory effects against at least one pathogenic bacterium. Conclusion This study has enriched the chemical diversity of secondary metabolites from the entomopathogenic fungus Samsoniella hepiali. -
图 1 化合物1～11 的结构
注：化合物1、2、9：酰胺类化合物；化合物3、4、8：甾醇类化合物；化合物5：脂肪酸类化合物；化合物6：脑苷脂类化合物；化合物7：糖苷类化合物；化合物10：腺苷类化合物；化合物11：糖类化合物。
Figure 1. Chemical structures of compounds 1～ 11
图 2 Samsoniella hepialid CDB9-31发酵提取物 HPLC 分析谱
Figure 2. The HPLC chromatogram analysis of Samsoniella hepialid CDB9-31 fermentation extract
表 1 Samsoniella hepiali CDB9-31代谢产物的抑菌活性
Table 1. Antimicrobial activities of secondary metabolites isolated from Samsoniella hepiali CDB9-31
指示菌 抑菌圈直径 （mm） 1 2 4 5 7 8 Kan Van Amp AmB B. subtilis ATCC 9372 - - - - - - # # 35.20 # S. aureus ATCC 6538 - - - - - - # # 20.00 # S. pneumoniae BNCC 338425 - - - - - 13.00 # # 21.20 # E. coli ATCC 25922 - - - - - - 16.30 # # # P. aeruginosa ATCC 15442 - - - - - - 25.00 # # # A. baumanii ATCC 19606 - - 8.00 - - - 18.30 # # # K. pneumoniae ATCC 13883 - 6.50 - - - - 15.10 # # # H. influenza ATCC 10211 - - 8.10 - - - 18.00 # # # S. typhimurium ATCC 14028 10.00 8.00 10.20 - 7.00 - 15.00 # # # MRSA 43300 - - - - - - # 16.00 # # C. albicans ATCC 14053 - - - 8.00 - - # # # 16.00 Drug-resistant C. albicans 23# - - - - - - # # # 10.00 注：抑菌圈直径包含滤纸片直径（6.00 mm）；“-”代表在给药剂量下无抗菌活性；“#”表示未进行该项测试；Kan：卡那霉素；Van：万古霉素；Amp：氨苄青霉素；AmB：两性霉素B。 
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表 2 化合物1～2、4～5和7～8的最小抑菌浓度
Table 2. The minimum inhibitory concentrations of compounds 1～ 2，4～ 5，and 7～ 8
指示菌 化合物MIC（μg/mL） 1 2 4 5 7 8 Kan Amp AmB S. pneumoniae BNCC 338425 # # # # # 128 # 8 # K. pneumoniae ATCC 13883 # 512 # # # # 64 # # H. influenza ATCC 10211 # # 128 # # # 32 # # S. typhimurium ATCC 14028 512 > # # > # 8 # # C. albicans ATCC 14053 # # # 512 # # # # 16 注：“>”代表MIC大于512 μg/mL；“#”表示未进行该项测试；Kan：卡那霉素；Amp：氨苄青霉素；AmB：两性霉素B。
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