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Quantitative proteomic profiling of Pseudomonas putida KT2440 in response to linalool and nerol stress

ABSTRACT: This project investigated the global proteomic response of Pseudomonas putida KT2440 to two representative monoterpenes, linalool and nerol, using data-independent acquisition (DIA)-based quantitative proteomics. The experiment comprised five treatment groups: a non-stressed control(CK), low and high inhibitory concentrations of linalool (0.81 g/L (LL)and 0.90 g/L(HL)), and low and high inhibitory concentrations of nerol (0.63 g/L(LN) and 0.81 g/L(HN)). Each group included three biological replicates. Cells were harvested 4 hours post-exposure. Differentially expressed proteins were identified according to the following criteria: proteins with a fold change (FC) greater than 1.5 and a p-value less than 0.05 were considered significantly up-regulated; conversely, proteins with a FC less than 0.67 and a p-value less than 0.05 were considered significantly down-regulated.. This dataset provides insights into the molecular mechanisms underlying the intrinsic monoterpene tolerance of P. putida KT2440 and supports the discovery of key genetic determinants for engineering robust microbial cell factories.

ORGANISM(S): Pseudomonas Putida Kt2440

SUBMITTER: Zhenmei Lu

PROVIDER: PXD074809 | iProX | Thu Feb 12 00:00:00 GMT 2026

REPOSITORIES: iProX

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Project description:Pyruvate kinase M2 (PKM2), as the terminal and last rate-limiting enzyme of the glycolytic pathway, is an ideal enzyme for regulating metabolic phenotype. PKM2 tetramer activation has shown a protective role against diabetic kidney disease (DKD). However, the molecular mechanisms involved in diabetic tubular has not been investigated so far. In this study, we performed transcriptome gene expression profiling in human renal proximal tubular epithelial cell line (HK-2 cells) treated with high D-glucose (HG) for 7 days before the addition of 10-μM TEPP-46, an activator of PKM2 tetramerization, for a further 1 day in the presence of HG. Afterwards, we analyzed the differentially expressed (DE) genes and investigated gene relationships based on weighted gene co-expression network analysis (WGCNA). The results showed that 2,902 DE genes were identified (adjusted P-value ≤ 0.05), where 2,509 DE genes (86.46%) were co-expressed in the key module. Four extremely down-regulated DE genes (HSPA8, HSPA2, HSPA1B and ARRB1) and three extremely up-regulated DE genes (GADD45A, IGFBP3 and SIAH1) enriched in the down-regulated endocytosis (hsa04144) and up-regulated p53 signaling pathway (hsa04115), respectively, were validated by the qRT-PCR experiments. The qRT-PCR results showed that the relative expression levels of HSPA8 (adjusted P-value = 4.45×10-34 & log2(FC) = -1.12), HSPA2 (adjusted P-value = 6.09×10-14 & log2(FC) = -1.27), HSPA1B (adjusted P-value = 1.14×10-11 & log2(FC) = -1.02) and ARRB1 (adjusted P-value = 2.60×10-5 & log2(FC) = -1.13) were significantly different (P-value < 0.05) from case group to control group. Furthermore, the interactions and predicted microRNAs of the key genes (HSPA8, HSPA2, HSPA1B and ARRB1) were visualized in networks. This study identified the key candidate transcriptomic biomarkers and biological pathways in the hyperglycemic HK-2 cells responding to the PKM2 activator TEPP-46. These results will be valuable for further research on PKM2 in DKD.

Dataset Information

Quantitative proteomic profiling of Pseudomonas putida KT2440 in response to linalool and nerol stress

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