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来自内生真菌链格孢的胆碱酯酶抑制剂(链格孢菌素):优化、纯化及特性研究

Cholinesterase inhibitor (Altenuene) from an endophytic fungus Alternaria alternata: optimization, purification and characterization.

作者信息

Bhagat J, Kaur A, Kaur R, Yadav A K, Sharma V, Chadha B S

机构信息

Department of Microbiology, Guru Nanak Dev University, Amritsar, Punjab, India.

Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India.

出版信息

J Appl Microbiol. 2016 Oct;121(4):1015-25. doi: 10.1111/jam.13192. Epub 2016 Aug 1.

Abstract

AIMS

The aim of this study was to screen endophytic fungi isolated from Vinca rosea for their potential to produce acetylcholinesterase (AChE) inhibitors.

METHOD AND RESULTS

Endophytic fungi isolated from V. rosea (Catharanthus roseus), were screened for AChE inhibitor production using Ellman's method. Maximum inhibition against AChE (78%) was observed in an isolate VS-10, identified to be Alternaria alternata on morphological and molecular basis. The isolate also inhibited butyrylcholinesterase (73%). Significant increase (1·3 fold) was achieved after optimization of process parameters using one variable at time approach. The inhibitor was purified using chromatographic techniques. The structure elucidation of the inhibitor was carried out using spectroscopic techniques and was identified to be 'altenuene'. The purified inhibitor possessed antioxidant potential as revealed by dot blot assay. The insecticidal potential of purified inhibitor was evaluated by feeding Spodoptora litura on diet amended with inhibitor. It evinced significant larval mortality.

CONCLUSIONS

Endophytic A. alternata can serve as a source of dual cholinesterase inhibitor 'altenuene' with significant antioxidant and insecticidal activity. This is the first report on acetylcholinestearse inhibitory activity of altenuene.

SIGNIFICANCE AND IMPACT OF THE STUDY

Alternaria alternata has the potential to produce a dual ChE inhibitor with antioxidant activity useful in the treatment of neurodegenerative disorders and in agriculture as biocontrol agent.

摘要

目的

本研究旨在筛选从长春花中分离出的内生真菌,以评估其产生乙酰胆碱酯酶(AChE)抑制剂的潜力。

方法与结果

采用Ellman法筛选从长春花中分离出的内生真菌产生AChE抑制剂的情况。在一株编号为VS - 10的分离株中观察到对AChE的最大抑制率(78%),通过形态学和分子学鉴定该分离株为链格孢菌。该分离株还抑制丁酰胆碱酯酶(73%)。采用一次只改变一个变量的方法优化工艺参数后,抑制率显著提高(1.3倍)。使用色谱技术对抑制剂进行纯化。利用光谱技术对抑制剂的结构进行解析,鉴定其为“细交链孢菌酮酸”。斑点印迹分析表明纯化后的抑制剂具有抗氧化潜力。通过用添加抑制剂的饲料喂养斜纹夜蛾来评估纯化后抑制剂的杀虫潜力。结果显示幼虫死亡率显著。

结论

内生链格孢菌可作为双胆碱酯酶抑制剂“细交链孢菌酮酸”的来源,该抑制剂具有显著的抗氧化和杀虫活性。这是关于细交链孢菌酮酸乙酰胆碱酯酶抑制活性的首次报道。

研究的意义和影响

链格孢菌有潜力产生一种具有抗氧化活性的双胆碱酯酶抑制剂,可用于治疗神经退行性疾病以及在农业中作为生物防治剂。

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