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从三叶番荔枝 Trichilia elegans A. JUSS ssp. elegans 中分离的内生真菌 Phomopsis longicolla 产生 3-硝基丙酸及其生物活性评价。

Production of 3-nitropropionic acid by endophytic fungus Phomopsis longicolla isolated from Trichilia elegans A. JUSS ssp. elegans and evaluation of biological activity.

机构信息

Laboratory of Microbial Biotechnology, Department of Biotechnology, Genetics and Cell Biology (DBC), State University of Maringá (UEM), Maringá, PR, Brazil.

出版信息

World J Microbiol Biotechnol. 2013 May;29(5):923-32. doi: 10.1007/s11274-013-1251-2. Epub 2013 Jan 8.

Abstract

The compound 3-nitropropionic acid is a potent neurotoxic agent in animals and well-known as a potent inhibitor of Mycobacterium tuberculosis. In this research, we were able to extract this compound from the endophytic fungus, Phomopsis longicolla (FJ62759), isolated from Trichilia elegans A. JUSS ssp. elegans. The aim of this study was the isolation of secondary metabolites produced by P. longicolla, the chemical identification of these compounds and evaluation of their antimicrobial and insecticidal activity. To accomplish these goals, the fungus was cultured in BD broth for 25 days without agitation at 28 °C, and then the broth was separated from the mycelium. The supernatant was partitioned with dichloromethane (CH2Cl2), ethyl acetate (EtOAc), and butanol (BuOH) solvents resulting in 3 extracts. However, only the EtOAc extract was used for fractionation and chemical identification because it had the greatest mass. After common chromatographic procedures, the fractions were analyzed by nuclear magnetic resonance to elucidate the chemical components. This procedure resulted in the identification of 3-nitropropionic acid in the D fraction. Evaluation of the insecticidal and antimicrobial activity of this compound has been accomplished, and the results indicate good inhibition of the citrus pathogen Guignardia citricarpa and cocoa pathogen Moniliophthora perniciosa and slight inhibition of the human bacterial pathogens Micrococcus luteus, Salmonella typhi and slight inhibition of phytopathogenic bacteria Xanthomonas axonopodis pv. phaseoli. The evaluation of insecticide activity did not show mortality of the Diatraea saccharalis larvae by the metabolite 3-nitropropionic acid in the D fraction. The results suggest that P. longicolla is a bioactive metabolic producing endophytic fungus with biotechnological properties.

摘要

3-硝基丙酸是一种有效的动物神经毒素,也是一种已知的结核分枝杆菌有效抑制剂。在这项研究中,我们从 Trichilia elegans A. JUSS ssp. elegans 内生真菌 Phomopsis longicolla (FJ62759) 中提取了这种化合物。本研究的目的是分离 P. longicolla 产生的次生代谢产物,鉴定这些化合物的化学结构,并评估它们的抗菌和杀虫活性。为了实现这些目标,将真菌在 BD 肉汤中于 28°C 下不搅拌培养 25 天,然后将肉汤与菌丝体分离。将上清液用二氯甲烷(CH2Cl2)、乙酸乙酯(EtOAc)和正丁醇(BuOH)溶剂萃取,得到 3 种提取物。然而,只有 EtOAc 提取物用于分馏和化学鉴定,因为它的质量最大。经过常见的色谱程序后,通过核磁共振分析对馏分进行分析,以阐明化学成分。这一程序在 D 馏分中鉴定出 3-硝基丙酸。已经完成了对这种化合物的杀虫和抗菌活性的评价,结果表明它对柑橘病原菌 Guignardia citricarpa 和可可病原菌 Moniliophthora perniciosa 具有良好的抑制作用,对人类细菌病原体 Micrococcus luteus、Salmonella typhi 有轻微的抑制作用,对植物病原菌 Xanthomonas axonopodis pv. phaseoli 有轻微的抑制作用。杀虫活性评价结果表明,D 馏分中的代谢产物 3-硝基丙酸对 Diatraea saccharalis 幼虫没有致死作用。结果表明,P. longicolla 是一种具有生物技术特性的生物活性代谢产内生真菌。

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