季也蒙毕赤酵母通过Hsf1p/PgSDR-A5D9S1降解展青霉素的分子机制研究

展青霉素污染已成为果汁行业安全生产的瓶颈。新近研究发现生物降解在展青霉素控制方面呈现出潜在的应用价值，而如何科学解析展青霉素降解机制是此类技术首要解决的科学问题。申请人前期筛选得到一株高效降解展青霉素的季也蒙毕赤酵母，并发现该酵母降解展青霉素的关键酶PgSDR-A5D9S1及其转录因子Hsf1p。然而，Hsf1p调控PgSDR-A5D9S1降解展青霉素的分子机制尚不明晰。为此，本项目通过分析展青霉素降解过程中PgSDR-A5D9S1的分子表达明确PgSDR-A5D9S1对展青霉素的降解作用；利用转录因子与启动子的结合实验探明Hsf1p对PgSDR-A5D9S1的转录调控作用；基于Hsf1p敲除菌株的多组学分析，解析Hsf1p下游基因分子表达与展青霉素降解的量效关系，揭示Hsf1p/PgSDR-A5D9S1降解展青霉素的分子机制。预期结果可为生物降解功能菌株产业化应用提供理论依据和技术支撑。

Patulin contamination has become a bottleneck problem to safety production of fruit juice at present. According to research findings, biodegradation technology shows potential application value in patulin control. However, clarifying the mechanism of patulin degradation scientifically is the principal science question for biodegradation technology. We previously screened a Pichia guilliermondii strain highly effective in patulin degradation by biodegradation technology, found the key patulin degradation enzyme PgSDR-A5D9S1 and its transcription factor Hsf1p. Nevertheless, the specific regulation mechanism of patulin degradation through the regulation of PgSDR-A5D9S1 with Hsf1p was not clear yet. In view of this, patulin degradation capability of PgSDR-A5D9S1 was verified by detecting molecular expression of PGSDR-A5D9S1 during the progress of patulin degradation. In addition, binding site analyzing assay between transcription factor Hsf1p and PgSDR-A5D9S1 promoter was conducted to elucidate the transcriptional regulation effect of Hsf1p on PgSDR-A5D9S1. Furthermore, the dose-effect relationship between patulin degradation and HSF1p downstream gene expression was analyzed to reveal the molecular mechanism of patulin degradation through the regulation of PgSDR-A5D9S1 with Hsf1p based on the multi-omics analysis of HSF1p knockout strains. The expected results of this project could provide theoretical basis and technical support for the industrial application of biodegradable functional yeast strains.