The long-chain flavodoxin FldX1 improves the biodegradation of 4-hydroxyphenylacetate and 3-hydroxyphenylacetate and counteracts the oxidative stress associated to aromatic catabolism in Paraburkholderia xenovorans.

IF 4.3 2区 生物学 Q1 BIOLOGY Biological Research Pub Date : 2024-04-01 DOI:10.1186/s40659-024-00491-4
Laura Rodríguez-Castro, Roberto E Durán, Valentina Méndez, Flavia Dorochesi, Daniela Zühlke, Katharina Riedel, Michael Seeger
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Abstract

Background: Bacterial aromatic degradation may cause oxidative stress. The long-chain flavodoxin FldX1 of Paraburkholderia xenovorans LB400 counteracts reactive oxygen species (ROS). The aim of this study was to evaluate the protective role of FldX1 in P. xenovorans LB400 during the degradation of 4-hydroxyphenylacetate (4-HPA) and 3-hydroxyphenylacetate (3-HPA).

Methods: The functionality of FldX1 was evaluated in P. xenovorans p2-fldX1 that overexpresses FldX1. The effects of FldX1 on P. xenovorans were studied measuring growth on hydroxyphenylacetates, degradation of 4-HPA and 3-HPA, and ROS formation. The effects of hydroxyphenylacetates (HPAs) on the proteome (LC-MS/MS) and gene expression (qRT-PCR) were quantified. Bioaugmentation with strain p2-fldX1 of 4-HPA-polluted soil was assessed, measuring aromatic degradation (HPLC), 4-HPA-degrading bacteria, and plasmid stability.

Results: The exposure of P. xenovorans to 4-HPA increased the formation of ROS compared to 3-HPA or glucose. P. xenovorans p2-fldX1 showed an increased growth on 4-HPA and 3-HPA compared to the control strain WT-p2. Strain p2-fldX1 degraded faster 4-HPA and 3-HPA than strain WT-p2. Both WT-p2 and p2-fldX1 cells grown on 4-HPA displayed more changes in the proteome than cells grown on 3-HPA in comparison to glucose-grown cells. Several enzymes involved in ROS detoxification, including AhpC2, AhpF, AhpD3, KatA, Bcp, CpoF1, Prx1 and Prx2, were upregulated by hydroxyphenylacetates. Downregulation of organic hydroperoxide resistance (Ohr) and DpsA proteins was observed. A downregulation of the genes encoding scavenging enzymes (katE and sodB), and gstA and trxB was observed in p2-fldX1 cells, suggesting that FldX1 prevents the antioxidant response. More than 20 membrane proteins, including porins and transporters, showed changes in expression during the growth of both strains on hydroxyphenylacetates. An increased 4-HPA degradation by recombinant strain p2-fldX1 in soil microcosms was observed. In soil, the strain overexpressing the flavodoxin FldX1 showed a lower plasmid loss, compared to WT-p2 strain, suggesting that FldX1 contributes to bacterial fitness. Overall, these results suggest that recombinant strain p2-fldX1 is an attractive bacterium for its application in bioremediation processes of aromatic compounds.

Conclusions: The long-chain flavodoxin FldX1 improved the capability of P. xenovorans to degrade 4-HPA in liquid culture and soil microcosms by protecting cells against the degradation-associated oxidative stress.

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长链黄酮毒素 FldX1 可改善 4-hydroxyphenylacetate 和 3-hydroxyphenylacetate 的生物降解,并抵消 Paraburkholderia xenovorans 中与芳香族分解代谢相关的氧化应激。
背景:细菌的芳香降解可能会导致氧化应激。Paraburkholderia xenovorans LB400 的长链黄酮毒素 FldX1 可抵消活性氧(ROS)。本研究的目的是评估 FldX1 在 Paraburkholderia xenovorans LB400 降解 4-hydroxyphenylacetate (4-HPA) 和 3-hydroxyphenylacetate (3-HPA) 过程中的保护作用:方法:在过表达 FldX1 的 P. xenovorans p2-fldX1 中评估了 FldX1 的功能。方法:在过量表达 FldX1 的 P. xenovorans p2-fldX1 中评估了 FldX1 的功能。研究了 FldX1 对 P. xenovorans 的影响,测量了其在羟基苯乙酸酯上的生长、4-HPA 和 3-HPA 的降解以及 ROS 的形成。还量化了羟基苯乙酸盐(HPAs)对蛋白质组(LC-MS/MS)和基因表达(qRT-PCR)的影响。用菌株 p2-fldX1 对受 4-HPA 污染的土壤进行生物增强评估,测量芳烃降解(HPLC)、4-HPA 降解细菌和质粒稳定性:结果:与 3-HPA 或葡萄糖相比,P. xenovorans 暴露于 4-HPA 会增加 ROS 的形成。与对照菌株 WT-p2 相比,P. xenovorans p2-fldX1 在 4-HPA 和 3-HPA 上的生长速度加快。与 WT-p2 菌株相比,p2-fldX1 菌株降解 4-HPA 和 3-HPA 的速度更快。与生长在葡萄糖中的细胞相比,生长在 4-HPA 上的 WT-p2 和 p2-fldX1 细胞比生长在 3-HPA 上的细胞显示出更多的蛋白质组变化。参与 ROS 解毒的几种酶,包括 AhpC2、AhpF、AhpD3、KatA、Bcp、CpoF1、Prx1 和 Prx2,在羟基苯乙酸盐的作用下上调。观察到有机过氧化氢抗性(Ohr)和 DpsA 蛋白下调。在 p2-fldX1 细胞中观察到编码清除酶(katE 和 sodB)、gstA 和 trxB 的基因下调,这表明 FldX1 阻止了抗氧化反应。在两种菌株在羟基苯乙酸盐上的生长过程中,包括孔蛋白和转运体在内的 20 多种膜蛋白的表达发生了变化。在土壤微生态系统中,重组菌株 p2-fldX1 对 4-HPA 的降解增加。在土壤中,与 WT-p2 菌株相比,过表达黄酮毒素 FldX1 的菌株的质粒损失较少,这表明 FldX1 对细菌的适应性有贡献。总之,这些结果表明,重组菌株 p2-fldX1 是一种有吸引力的细菌,可用于芳香族化合物的生物修复过程:结论:长链黄酮毒素 FldX1 通过保护细胞免受与降解相关的氧化应激,提高了 P. xenovorans 在液体培养和土壤微生态系统中降解 4-HPA 的能力。
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来源期刊
Biological Research
Biological Research 生物-生物学
CiteScore
10.10
自引率
0.00%
发文量
33
审稿时长
>12 weeks
期刊介绍: Biological Research is an open access, peer-reviewed journal that encompasses diverse fields of experimental biology, such as biochemistry, bioinformatics, biotechnology, cell biology, cancer, chemical biology, developmental biology, evolutionary biology, genetics, genomics, immunology, marine biology, microbiology, molecular biology, neuroscience, plant biology, physiology, stem cell research, structural biology and systems biology.
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