Differential protein expression during growth on model and commercial mixtures of naphthenic acids in Pseudomonas fluorescens Pf-5,MicrobiologyOpen

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Differential protein expression during growth on model and commercial mixtures of naphthenic acids in Pseudomonas fluorescens Pf-5
MicrobiologyOpen ( IF 3.9 ) Pub Date : 2021-07-19 , DOI: 10.1002/mbo3.1196
Boyd A McKew ₁ , Richard Johnson ₁ , Lindsay Clothier _{2,

3} , Karl Skeels ₁ , Matthew S Ross ₄ , Metodi Metodiev ₁ , Max Frenzel ₅ , Lisa M Gieg ₃ , Jonathan W Martin ₆ , Michael A Hough ₁ , Corinne Whitby ₁

Affiliation

Naphthenic acids (NAs) are carboxylic acids with the formula (C_nH_2n₊_ZO₂) and are among the most toxic, persistent constituents of oil sands process-affected waters (OSPW), produced during oil sands extraction. Currently, the proteins and mechanisms involved in NA biodegradation are unknown. Using LC-MS/MS shotgun proteomics, we identified proteins overexpressed during the growth of Pseudomonas fluorescens Pf-5 on a model NA (4′-n-butylphenyl)-4-butanoic acid (n-BPBA) and commercial NA mixture (Acros). By day 11, >95% of n-BPBA was degraded. With Acros, a 17% reduction in intensity occurred with 10–18 carbon compounds of the Z family −2 to −14 (major NA species in this mixture). A total of 554 proteins (n-BPBA) and 631 proteins (Acros) were overexpressed during growth on NAs, including several transporters (e.g., ABC transporters), suggesting a cellular protective response from NA toxicity. Several proteins associated with fatty acid, lipid, and amino acid metabolism were also overexpressed, including acyl-CoA dehydrogenase and acyl-CoA thioesterase II, which catalyze part of the fatty acid beta-oxidation pathway. Indeed, multiple enzymes involved in the fatty acid oxidation pathway were upregulated. Given the presumed structural similarity between alkyl-carboxylic acid side chains and fatty acids, we postulate that P. fluorescens Pf-5 was using existing fatty acid catabolic pathways (among others) during NA degradation.

中文翻译：

荧光假单胞菌 Pf-5 模型和商业环烷酸混合物生长过程中的差异蛋白表达

环烷酸 (NAs) 是分子式为 (C _n H _2n ₊ _Z O ₂ ) 的羧酸，是油砂开采过程中产生的油砂工艺影响水 (OSPW) 中毒性最强、最持久的成分之一。目前，参与 NA 生物降解的蛋白质和机制尚不清楚。使用 LC-MS/MS 鸟枪法蛋白质组学，我们鉴定了荧光假单胞菌Pf-5 在模型 NA (4'- n-丁基苯基)-4-丁酸 ( n -BPBA) 和商业 NA 混合物 (Acros) 上生长过程中过表达的蛋白质）。到第 11 天，>95% 的n -BPBA 被降解。对于 Acros， Z家族 -2 至 -14（该混合物中的主要 NA 物种）的 10-18 碳化合物的强度降低了 17%。在 NA 上生长期间，总共有 554 个蛋白质 ( n -BPBA) 和 631 个蛋白质 (Acros) 过表达，包括几种转运蛋白（例如 ABC 转运蛋白），表明细胞对 NA 毒性有保护性反应。与脂肪酸、脂质和氨基酸代谢相关的几种蛋白质也过度表达，包括酰基辅酶A脱氢酶和酰基辅酶A硫酯酶II，它们催化部分脂肪酸β-氧化途径。事实上，参与脂肪酸氧化途径的多种酶都被上调。鉴于烷基羧酸侧链和脂肪酸之间的假定结构相似性，我们假设P 。荧光菌Pf-5 在 NA 降解过程中利用现有的脂肪酸分解代谢途径（以及其他途径）。

更新日期：2021-07-20

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