BIOACTIVE INDOLE DITERPENOIDS AND POLYKETIDES FROM THE MARINE-DERIVED FUNGUS Penicillium javanicum
BIOACTIVE INDOLE DITERPENOIDS AND POLYKETIDES FROM THE MARINE-DERIVED FUNGUS Penicillium javanicum
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来自海洋源性真菌爪哇青霉的生物活性吲哚二萜和聚酮化合物
DOI：
10.1007/s10600-020-03039-6
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发表时间：
2020-04-01
影响因子：
0.8
通讯作者：
Wang, Chang-Yun
中科院分区：
文献类型：
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作者：
Liang, Zhao-Yang;Shen, Nan-Xing;Wang, Chang-Yun
红树林相关微生物，发生在热带和亚热带潮间带河口区，已被证明是结构独特和生物活性天然产物的丰富来源，可用于开发新的药物[1，2]。在我们对中国南海红树林相关真菌的持续研究中，我们发现了几种具有抗菌活性的新代谢产物[3]。最近，一株红树林根际土壤来源的真菌Penicillium javanicum HK 1 -23被选中进行化学研究，因为其提取物具有抑菌活性并且具有有趣的HPLC-UV特征。真菌粗提物的进一步分级分离导致发现七种吲哚二萜（1-7）和两种聚酮化合物（8，9）。2015年9月，从海南岛东寨港红树林自然保护区的红树林根际土壤中分离到一株爪哇假单胞菌（P. javanicum HK 1 -23）。该菌株保藏在中国扬州大学环境科学与工程学院海洋科学与技术研究所。根据其形态特征和分子协议，通过扩增和测序的DNA的ITS区的rRNA基因的真菌进行鉴定。经鉴定，该真菌为爪哇青霉（P. javanicum），其591 bp的ITS序列与爪哇青霉（P. javanicum）（MH 877078）的ITS序列同源性为100%。1）。该序列已提交到GenBank，登录号为MH 628211。将真菌菌株在马铃薯葡萄糖肉汤培养基（20 g葡萄糖和30 g人造海盐在1 L马铃薯浸液中; 1 L锥形瓶，每个锥形瓶含有400 mL培养肉汤）中在室温下不振荡培养4周。提取和分离。过滤发酵液（30 L）以将发酵液与菌丝体分离。然后将肉汤用等体积的EtOAc萃取三次，并将菌丝体用MeOH萃取三次。将有机萃取物合并并在真空下浓缩，得到总萃取物（20.0g），将其进行硅胶柱色谱法（CC），使用EtOAc-石油醚（0-100%），然后用MeOH-EtOAc（0-100%）的分步梯度洗脱，分离成八个级分（Frs.第1-8段）。通过重复硅胶CC和半制备型HPLC分离和纯化亚级分，得到1（25 mg）、2（3 mg）、3（12 mg）、4（34 mg）、5（20 mg）、6（10 mg）、7（4 mg）、8（30 mg）和9（100 mg）。
Mangrove-associated microorganisms, occurring at the tropical and subtropical intertidal estuarine zones, have proven to be a prolific source of structurally unique and biologically active natural products that could be used for the development of new medicinal agents [1, 2]. In our ongoing research on mangrove-associated fungi from the South China Sea, we have found several new metabolites with antibacterial activity [3]. Recently, a mangrove rhizosphere soil-derived fungus strain Penicillium javanicum HK1-23 was selected to be chemically investigated because its extracts showed antibacterial activity and had an interesting HPLC-UV profile. Further fractionation of the fungal crude extracts led to the discovery of seven indole diterpenes (1–7) and two polyketides (8, 9). Herein we report the isolation, structure elucidation, and antibacterial activities of these compounds.The fungus P. javanicum HK1-23 was isolated from mangrove rhizosphere soil collected from Dongzhaigang mangrove natural reserve in Hainan Island in September 2015. The strain was deposited at the Marine Science & Technology Institute, College of Environmental Science & Engineering, Yangzhou University, Yangzhou, PR China. The fungus was identified according to its morphological traits and a molecular protocol by amplification and sequencing of the DNA of the ITS region of the rRNA gene. The fungus was identified as a P. javanicum, whose 591 base pair ITS sequence had 100% sequence identity to that of P. javanicum (MH877078. 1). The sequence data have been submitted to GenBank with accession number MH628211. The fungal strain was cultivated in potato dextrose broth medium (20 g of glucose and 30 g of artificial seasalt in 1 L of potato infusion; 1 L Erlenmeyer flasks each containing 400 mL of culture broth) at room temperature without shaking for 4 weeks. Extraction and Isolation. The fermentation broth (30 L) was filtered to separate the broth from the mycelia. Then the broth was extracted three times with an equal volume of EtOAc, and the mycelia were extracted three times with MeOH. The organic extracts were combined and concentrated under vacuum to afford a total extract (20.0 g), which was subjected to silica gel column chromatography (CC) using step gradient elution with EtOAc–petroleum ether (0–100%) and then with MeOH–EtOAc (0–100%) for separation into eight fractions (Frs. 1–8). The subfractions were isolated and purified by repeated silica gel CC and semi-preparative HPLC to obtain 1 (25 mg), 2 (3 mg), 3 (12 mg), 4 (34 mg), 5 (20 mg), 6 (10 mg), 7 (4 mg), 8 (30 mg), and 9 (100 mg).