一株西藏卡里比克迈耶氏酵母对玉米赤霉烯酮的降解效果及作用机制研究

    Research on the Degradation Efficacy and Mechanism of Zearalenone by Tibetan Yeast Meyerozyma caribbica

    • 摘要:
      目的 从西藏土壤中分离筛选出一株具有玉米赤霉烯酮(Zearalenone, ZEN)降解能力的酵母菌株, 并研究其在不同条件下的降解特性及降解机制, 为生物降解毒素提供理论依据。
      方法 从西藏土壤样品中分离并筛选具有ZEN降解能力的菌株,进一步筛选ZEN降解能力最强的菌株68进行分析。通过26S rDNA基因测序与系统进化树分析, 对菌株68进行鉴定, 研究其在菌液浓度分别为1×106、1×107、1×108、1×109个/mL,及不同菌液处理(活酵母细胞、热杀死酵母细胞、酵母上清液、酵母内含物)对ZEN降解能力的影响; 并在降解48 h时对试验菌种取样进行转录组学分析,解析毒素降解发挥作用相关通路及基因。
      结果 从西藏土壤样品中筛选出1株对ZEN降解效果较好的菌株,命名为菌株68, 初步验证其对ZEN的降解率达70%以上;通过26S rDNA基因测序与系统进化树分析,鉴定菌株68为卡里比克迈耶氏酵母(Meyerozyma caribbica)。菌株68降解ZEN的最适菌液浓度为1×108个/mL,降解率达80.3%。菌株68接种后活细胞在24 h内对ZEN的降解率升至31.67%,72 h后达72.93%;细胞外上清液24 h降解率为12.96%,72 h后约15%;细胞内含物24 h降解率为4.36%,72 h后为26.96%;热杀死酵母细胞72 h内降解率基本为0。不同菌液处理试验结果表明,该酵母对ZEN的降解机制先表现为吸附作用,再在胞内进行生物降解作用。转录组学结果表明,在GO富集分析中富集差异表达基因(Differentially Expressed Genes,DEGs)集中在氧化还原酶活性、参与醛或羰基供体反应的氧化还原酶活性;KEGG富集分析表明,糖酵解/糖异生显著富集到13个通路,其中与氧化还原相关通路显著富集上调,进一步明确毒素降解受氧化还原机制影响。
      结论 菌株68可通过胞内氧化还原酶系统高效降解ZEN,其分子机制与能量代谢驱动的辅因子NADH/NADPH供应密切相关。

       

      Abstract:
      Objective To isolate and screen a yeast strain capable of degrading zearalenone (ZEN) from Tibetan soil samples and investigate its degradation characteristics and mechanisms under varying conditions, thereby providing a theoretical basis for mycotoxin detoxification.
      Method Yeast strains were isolated from Tibetan soil samples, and strains with potential zearalenone degradation activity were selected for further analysis.Strain 68 was taxonomically identified through 26S rDNA gene sequence analysis and phylogenetic tree construction, and its degradation capacity was evaluated under cell concentrations ranging from 1×106 to 1×109 cells/mL. Experimental treatments included viable cells, heat-killed cells, cell-free supernatant, and intracellular extracts.Samples were collected at 48 h for transcriptomic sequencing to predict degradation-related pathways and genes.
      Result A yeast strain designated 68, showing ZEN degradation rate over 70%, was isolated from Tibetan soil samples and identified as Meyerozyma caribbica.Optimal degradation conditions were determined as a cell concentration of 1×108 cells/mL, achieving an degradation rate of 80.3% after 72 h. Viable cells exhibited progressive degradation, with degradation rates reaching 31.67% at 24 h and 72.93% at 72 h.The cell-free supernatant showed limited activity, with degradation rates reaching 12.96% at 24 h and approximately 15% at 72 h, while intracellular extracts degraded 26.96% of ZEN by 72 h.Heat-killed cells displayed no degradation. Mechanistic studies revealed that ZEN degradation by strain 68 primarily involved initial adsorption followed by intracellular biodegradation.Transcriptomic analysis demonstrated that differentially expressed genes were significantly enriched in oxidoreductase activity and oxidoreductase activity, acting on the aldehyde or oxo group of donors.KEGG enrichment highlighted 13 sub-pathways in glycolysis/gluconeogenesis, with redox-associated pathways showing marked upregulation, confirming the critical role of oxidoreduction mechanisms in ZEN detoxification.
      Conclusion Strain 68 efficiently degrades ZEN via an intracellular oxidoreductase system, mediated by energy metabolism-dependent NADH/NADPH cofactor regeneration.This study provides a foundation for developing yeast-based biocontrol strategies to mitigate mycotoxin contamination in food and feed systems.

       

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