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iTRAQ-Based Quantitative Proteomic Analysis Reveals Changes in Metabolite Biosynthesis in Monascus purpureus in Response to a Low-Frequency Magnetic Field

机译:基于iTRAQ的定量蛋白质组学分析揭示了响应低频磁场的红曲霉代谢产物生物合成的变化

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摘要

Background: Low-frequency magnetic fields (LF-MFs) dampen the citrinin output by Monascus purpureus in fermentations. The influence of LF-MFs on biosynthesis by M. purpureus was evaluated at the protein level. Methods: Cultures were treated with a 1.6-mT MF from day 0 to day 2 of incubation, and secondary metabolite production was evaluated on the day 12 of incubation. All proteins were extracted from M. purpureus mycelia and subjected to isobaric tags for relative and absolute quantification (iTRAQ) labeling and subsequent liquid chromatography/mass spectrometry (LC-MS/MS) analysis on day 6 of fermentation. Results: There was no difference in biomass between the treated samples and the control. Citrinin production was 46.7% lower, and the yields of monacolin K and yellow, orange, and red pigment were 29.3%, 31.3%, 41.7%, and 40.3% higher, respectively, in the exposed samples compared to the control. Protein expression in M. purpureus under LF-MF treatment was quantified using iTRAQ technology. Of 2031 detected proteins, 205 were differentially expressed. The differentially-expressed proteins were subjected to Gene Ontology (GO) functional annotation and statistical analysis, which revealed that they mainly refer to biological metabolism, translation, antioxidant, transport and defense pathways. Among all the tagged proteins, emphasis was placed on the analysis of those involved in the synthesis of citrinin, pigment and monacolin K was emphasized. Conclusions: LF-MFs affected Monascus secondary metabolism at the protein level, and aggregate data for all the protein profiles in LF-MF-treated Monascus was obtained.
机译:背景:低频磁场(LF-MFs)抑制了红曲霉在发酵中输出的桔霉素。在蛋白质水平上评估了LF-MF对紫癜支原体生物合成的影响。方法:从培养的第0天到第2天,将培养物用1.6 mT MF处理,并在培养的第12天评估次生代谢产物的产生。从紫癜支原体菌丝体中提取所有蛋白,并在同等的标签上进行相对和绝对定量(iTRAQ)标记,并在发酵的第6天进行液相色谱/质谱(LC-MS / MS)分析。结果:处理后的样品和对照之间的生物量没有差异。与对照相比,在暴露的样品中,Citrinin的产量降低了46.7%,莫纳可林K以及黄色,橙色和红色颜料的产率分别提高了29.3%,31.3%,41.7%和40.3%。使用iTRAQ技术对LF-MF处理下紫癜支原体中的蛋白质表达进行定量。在2031个检测到的蛋白质中,有205个差异表达。对差异表达的蛋白质进行了基因本体论(GO)功能注释和统计分析,发现它们主要涉及生物代谢,翻译,抗氧化剂,转运和防御途径。在所有标记的蛋白质中,重点放在分析涉及柠檬绿素,色素和莫纳可林K的蛋白质上。结论:LF-MFs在蛋白质水平上影响红曲霉的二次代谢,并获得了LF-MF处理的红曲霉的所有蛋白质谱的汇总数据。

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