Optimization and Characterization of Flavipin Produced by Aspergillus Terreus

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Abstract

The secondary metabolites of microorganisms serve as defence or signalling molecules in ecological interactions, revealing substantial survival benefits in nature. As a result, many researchers have concentrated on screening and optimizing the production of these molecules from natural sources such as microorganisms with the objective of pharmacological uses, primarily as antibiotics or anticancer agents. In this study, 80 isolates of Aspergillus were investigated for the production of flavipin. These fungi were collected from various locations and laboratories. Flavipin was estimated by using a standard curve, then purified by using silica gel chromatography, followed by identification using thin layer chromatography (TLC), and High Performance liquid chromatography (HPLC). The fermentation conditions were carried out at the Central Health Laboratory/Maysan Health Directorate from April 2021 to August 2022. Out of eighty isolates of Aspergillus, only one isolate was identified as producer of flavipin which was Aspergillus terreus. According to HPLC analysis, the retention times of flavipin and its standard were 7.7 minutes and 7.6 minutes, respectively. By using the TLC technique, the relative flow (Rf) value was 0.55 cm for both standard flavipin and flavipin. The optimization of growth conditions and production of flavipin were studied. It is revealed that optimum conditions were as follows: pH 7 on 16 days, the temperature of 25oC for 12 days, culture volume of 50 ml on the 16th day, shaking speed of 150 rpm on the 12th day, inoculum size of 8 fungal agar disc on the 12th day, the optimal incubation period of 14 days, and Potato Dextrose Broth as the optimal culture media.  The aim of the study was to determination of optimal conditions for the flavipin production that produced by Aspergillus terreus. For yielding a profuse amount of flavipin, the incubation and fermentation conditions such as temperature, the culture volume, shaking speed, inoculum size, pH of the medium, incubation period, and the type of culture media should be considered and the optimal one must be chosen.
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地曲霉产黄素的优化与表征
微生物的次生代谢物在生态相互作用中充当防御或信号分子,揭示了自然界中大量的生存益处。因此,许多研究人员集中在筛选和优化这些分子的生产,从自然来源,如微生物的目的是药理学用途,主要是作为抗生素或抗癌剂。本研究对80株曲霉进行了产黄素的研究。这些真菌是从不同地点和实验室收集的。采用标准曲线法对黄酮类化合物进行预估,硅胶层析纯化,薄层色谱(TLC)和高效液相色谱(HPLC)鉴定。发酵条件于2021年4月至2022年8月在中央卫生实验室/Maysan卫生局进行。在80株分离的曲霉中,只有一株被鉴定为黄黄素的产生者,即地曲霉。HPLC法测定黄黄素的保留时间为7.7 min,黄黄素标准品的保留时间为7.6 min。薄层色谱法测定,标准黄素和黄素的相对流量(Rf)值均为0.55 cm。对黄素的生长条件和产量进行了优化研究。结果表明,最佳培养条件为:pH 7 16天,温度25℃12天,第16天培养量50 ml,第12天摇速150 rpm,第12天接种8个真菌琼脂盘,最佳培养时间为14天,以马铃薯葡萄糖肉汤为最佳培养基。本研究旨在确定地曲霉产黄素的最佳工艺条件。为获得大量黄素,应综合考虑温度、培养量、摇速、接种量、培养基pH、培养时间、培养基种类等培养发酵条件,选择最佳的黄素发酵条件。
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