Top-Down Characterization of an Antimicrobial Sanitizer, Leading From Quenchers of Efficacy to Mode of Action.
Top-Down Characterization of an Antimicrobial Sanitizer, Leading From Quenchers of Efficacy to Mode of Action.
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DOI：
10.3389/fmicb.2020.575157
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发表时间：
2020
影响因子：
5.2
通讯作者：
Avery SV
中科院分区：
文献类型：
--
作者：
Wohlgemuth F;Gomes RL;Singleton I;Rawson FJ;Avery SV
我们开发了一种自上而下的策略来表征抗菌氧化消毒剂，该消毒剂具有多种拟议应用，包括新鲜食品的表面消毒，并具有水弹性的好处。该策略涉及找到抗微生物活性的猝灭剂，然后通过从复杂基质开始识别关键化学反应伙伴，缩小对细胞内特定有机分子的反应性，找到抗微生物作用模式。消毒剂电解水（EW）保留部分杀菌活性对食品腐败真菌尼日尔在高水平的添加土壤（30-750毫克毫升-1），通常与收获的产品。土壤有机负荷高（98毫克g-1）得到更强的EW灭活。由复杂的有机物混合物（YEPD培养基）标记的灭活与其富含蛋白质的组分有关。添加纯蛋白质或氨基酸（≤1 mg mL-1）完全抑制EW活性。机制进一步询问与酵母模型，证实了显着抑制EW行动的氨基酸蛋氨酸。用蛋氨酸预培养可增加对EW、次氯酸钠或无氯臭氧化水的抗性。过表达蛋氨酸亚砜还原酶（还原氧化蛋氨酸）保护免受EW。基于荧光探针的分析表明，蛋氨酸和半胱氨酸取代了EW中的游离氯物种。细胞内蛋氨酸氧化可以干扰细胞的FeS-集群，我们表明，EW处理损害FeS-酶活性。该研究确立了自上而下的方法对消毒剂功效和作用的多层次表征的价值。结果揭示了蛋白质和氨基酸作为EW活性的关键淬灭剂，并且在氨基酸中，蛋氨酸氧化和FeS簇损伤对于抗微生物作用模式的重要性。
We developed a top-down strategy to characterize an antimicrobial, oxidizing sanitizer, which has diverse proposed applications including surface-sanitization of fresh foods, and with benefits for water resilience. The strategy involved finding quenchers of antimicrobial activity then antimicrobial mode of action, by identifying key chemical reaction partners starting from complex matrices, narrowing down reactivity to specific organic molecules within cells. The sanitizer electrolyzed-water (EW) retained partial fungicidal activity against the food-spoilage fungus Aspergillus niger at high levels of added soils (30–750 mg mL–1), commonly associated with harvested produce. Soil with high organic load (98 mg g–1) gave stronger EW inactivation. Marked inactivation by a complex organics mix (YEPD medium) was linked to its protein-rich components. Addition of pure proteins or amino acids (≤1 mg mL–1) fully suppressed EW activity. Mechanism was interrogated further with the yeast model, corroborating marked suppression of EW action by the amino acid methionine. Pre-culture with methionine increased resistance to EW, sodium hypochlorite, or chlorine-free ozonated water. Overexpression of methionine sulfoxide reductases (which reduce oxidized methionine) protected against EW. Fluoroprobe-based analyses indicated that methionine and cysteine inactivate free chlorine species in EW. Intracellular methionine oxidation can disturb cellular FeS-clusters and we showed that EW treatment impairs FeS-enzyme activity. The study establishes the value of a top-down approach for multi-level characterization of sanitizer efficacy and action. The results reveal proteins and amino acids as key quenchers of EW activity and, among the amino acids, the importance of methionine oxidation and FeS-cluster damage for antimicrobial mode-of-action.
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影响因子：
2.7
作者：
Benjamini, Yoav;Krieger, Abba M.;Yekutieli, Daniel
通讯作者：
Yekutieli, Daniel
影响因子：
3.3
作者：
Alhebshi A;Sideri TC;Holland SL;Avery SV
通讯作者：
Avery SV
影响因子：
3.9
作者：
Delaye, Luis;Becerra, Arturo;Lazcano, Antonio
通讯作者：
Lazcano, Antonio
DOI：
10.1007/s00775-018-1548-6
发表时间：
2018-06
期刊：
Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry
影响因子：
--
作者：
Cardenas-Rodriguez M;Chatzi A;Tokatlidis K
通讯作者：
Tokatlidis K
影响因子：
5.6
作者：
Gil, Maria I.;Gomez-Lopez, Vicente M.;Allende, Ana
通讯作者：
Allende, Ana