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一种基于聚酮合酶I基因的筛选方法,用于从橘青霉Salicorn 46中发现一种新的聚酮化合物。

A PKS I gene-based screening approach for the discovery of a new polyketide from Penicillium citrinum Salicorn 46.

作者信息

Wang Xiaomin, Wang Hui, Liu Tianxing, Xin Zhihong

机构信息

Key Laboratory of Food Processing and Quality Control, College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2014 Jun;98(11):4875-85. doi: 10.1007/s00253-014-5572-3. Epub 2014 Feb 18.

Abstract

Salicorn 46, an endophytic fungus isolated from Salicornia herbacea Torr., was identified as Penicillium citrinum based on its internal transcribed spacer and ribosomal large-subunit DNA sequences using a type I polyketide synthase (PKS I) gene screening approach. A new polyketide, penicitriketo (1), and seven known compounds, including ergone (2), (3β,5α,8α,22E)-5,8-epidioxyergosta-6,9,22-trien-3-ol (3), (3β,5α,8α,22E)-5,8-epidioxyergosta-6,22-dien-3-ol (4), stigmasta-7,22-diene-3β,5α,6α-triol (5), 3β,5α-dihydroxy-(22E,24R)-ergosta-7,22-dien-6β-yl oleate (6), N b-acetyltryptamine (7), and 2-(1-oxo-2-hydroxyethyl) furan (8), were isolated from the culture of Salicorn 46, and their chemical structures were elucidated by spectroscopic analysis. Antioxidant experiments revealed that compound 1 possessed moderate DPPH radical scavenging activity with an IC50 value of 85.33 ± 1.61 μM. Antimicrobial assays revealed that compound 2 exhibited broad-spectrum antimicrobial activity against Candida albicans, Clostridium perfringens, Mycobacterium smegmatis, and Mycobacterium phlei with minimal inhibitory concentration (MIC) values of 25.5, 25.5, 18.5, and 51.0 μM, respectively. Compound 3 displayed potent antimicrobial activities against C. perfringens and Micrococcus tetragenus with a MIC value of 23.5 μM. Compounds 5 and 6 showed high levels of selectivity toward Bacillus subtilis and M. phlei with MIC values of 22.5 and 14.4 μM, respectively. The results of this study highlight the use of PCR-based techniques for the screening of new polyketides from endophytic fungi containing PKS I genes.

摘要

盐角草46是从盐角草中分离出的一种内生真菌,基于其内部转录间隔区和核糖体大亚基DNA序列,采用I型聚酮合酶(PKS I)基因筛选方法,被鉴定为桔青霉。从盐角草46的培养物中分离出一种新的聚酮化合物——青霉三酮(1)和七种已知化合物,包括麦角甾醇(2)、(3β,5α,8α,22E)-5,8-环氧麦角甾-6,9,22-三烯-3-醇(3)、(3β,5α,8α,22E)-5,8-环氧麦角甾-6,22-二烯-3-醇(4)、豆甾-7,22-二烯-3β,5α,6α-三醇(5)、3β,5α-二羟基-(22E,24R)-麦角甾-7,22-二烯-6β-油酸酯(6)、N-乙酰色胺(7)和2-(1-氧代-2-羟乙基)呋喃(8),并通过光谱分析阐明了它们的化学结构。抗氧化实验表明,化合物1具有中等的DPPH自由基清除活性,IC50值为85.33±1.61μM。抗菌试验表明,化合物2对白色念珠菌、产气荚膜梭菌、耻垢分枝杆菌和草分枝杆菌具有广谱抗菌活性,最小抑菌浓度(MIC)值分别为25.5、25.5、18.5和51.0μM。化合物3对产气荚膜梭菌和四联微球菌具有较强的抗菌活性,MIC值为23.5μM。化合物5和6对枯草芽孢杆菌和草分枝杆菌表现出高度的选择性,MIC值分别为22.5和14.4μM。本研究结果突出了基于PCR技术在筛选含有PKS I基因的内生真菌中的新聚酮化合物方面的应用。

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