The development of novel cholesterol biosynthesis inhibitors is a task of major concern due to the diverse roles of cholesterol and its precursors in physiological processes. Therefore, appropriate screening assays are required, which can be used to identify and quantify specific inhibitors targeting the desired enzyme. Here, we developed a whole‐cell screening assay based on a HL60 cell line, which can be used to characterize inhibitors interacting with enzymes of the isoprenoid part of cholesterol biosynthesis. Due to the change of the isoprenoid pattern under enzyme inhibition, an identification of the targeted enzyme is possible. With the described assay, we can distinguish between free and pyrophosphorylated isoprenoids after enzymatic cleavage in cellular and extracellular matrices. The approach was validated in line with the European Medicines Agency guideline on bioanalytical method validation. As proof of concept, literature‐described inhibitors of the isoprenoid pathway were tested. We characterized the effect of 11 isoprenoid biosynthesis inhibitors, and we identified 6‐fluoromevalonate as an isopentenyl pyrophosphate isomerase inhibitor, a biological activity that was previously unknown. Furthermore, isoprenoid patterns revealed that, independent of the analyzed matrix, the predominant form of the detected isoprenoids were dephosphorylated isoprenoids and only small amounts were present as pyrophosphates.

2025-01-01·INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY

An atoxigenic Aspergillus flavus PA67 from Shandong province exhibits potential in biocontrol against toxigenic Aspergillus flavus, Sclerotium rolfsii, and Fusarium proliferatum

Article

作者: Zhang, Yanyan ; Ren, Junhe ; Xu, Jia ; Kong, Qing

Peanuts and corn are susceptible to various soil-borne fungi, leading to significant economic losses. Atoxigenic Aspergillus flavus have been widely used as biocontrol agents for managing aflatoxin contamination because of their minimal environmental impact, strong competitive ability, and sustained inhibition effect. After multiple identifications and cluster amplification pattern (CAP) analysis, three atoxigenic A. flavus PA04, PA10 and PA67 were isolated from peanut samples in Shandong Province, which can reduce aflatoxin levels by up to 90 %. Our study revealed that atoxigenic A. flavus also competed vigorously with Sclerotium rolfsii and Fusarium proliferatum for nutrition and space, achieving notable inhibition rates of up to 90.4 % and 90.6 %, respectively. The supernatants of atoxigenic A. flavus also inhibited the growth of S. rolfsii and F. proliferatum, with PA67 demonstrating the most significant effect. Whole genome sequencing revealed that PA67 contains multiple glycoside hydrolases and metabolites with antifungal activity. The kojic acid production of PA67 was higher than that of PA04 and PA10, reaching 17.48 g/L, which has a significant inhibition on sclerotia germination. PA67 supernatant significantly inhibited the hyphae growth of S. rolfsii and F. proliferatum, and down-regulated genes related to sclerotia and fumonisin formation. This study demonstrates the biocontrol potential of PA67 against three soil-borne fungi and is the first investigation of atoxigenic A. flavus to inhibit S. rolfsii and F. proliferatum.

2024-09-01·LWT-FOOD SCIENCE AND TECHNOLOGY

Transcriptomic analysis of the metabolic characteristics of Aspergillus oryzae during the fermentation of doenjang-meju, a traditional Korean fermented soybean brick

作者: Han, Dong Min ; Jeon, Che Ok ; Choi, Dae Gyu ; Baek, Ju Hye

To investigate the fermentative traits of Aspergillus oryzae in doenjang-meju, a traditional Korean fermented soybean brick, we prepared doenjang-meju bricks inoculated with both A. oryzae and Bacillus velezensis--recognized as crucial microbes in doenjang-meju fermentation, and then the transcriptomes of A. oryzae were analyzed throughout the entire fermentation period.The transcriptomic anal. of carbohydrate-active enzyme (CAZy) genes revealed that A. oryzae exhibited robust expression of CAZy genes involved in the decomposition of various carbohydrates, particularly cellulose, starch, and hemicellulose, indicating the active breakdown of soybean carbohydrates by A. oryzae during fermentationFurthermore, KEGG anal., metabolic pathway reconstruction, and their transcriptomic anal. provided a comprehensive overview of the decomposition patterns of various carbohydrates, lipids, and proteins and the metabolic features of various carbon compounds during fermentationAddnl., the transcriptomic anal. of biosynthetic gene clusters indicated the potential production of various secondary metabolites by A. oryzae during fermentationGenes related to carbohydrate and carbon compound metabolisms and secondary metabolite biosynthesis displayed unique and different transcriptional expression patterns depending on the specific gene and fermentation stage.Overall, our findings provide comprehensive insights into the metabolic characteristics of A. oryzae, thus contributing to our understanding of soybean fermentation

100 项与 Squalestatin 1 相关的药物交易

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