Biobutanol has a high energy density, good miscibility with gasoline, and a low environmental impact; therefore, it is a possible substitute or additive for gasoline. Because biobutanol is typically obtained through so-called ABE fermentation, the effective separation of butanol from the product mixture is important. Ionic liquids (ILs) have been used as extractants, and tailoring the IL structure to promote hydrogen bonding with butanol could improve the extraction performance. In the present work, an amine group was added to the imidazole cation of an IL, paired with bis(trifluoromethylsulfonyl)imide as the anion. Hydroxyl groups were added to the 1,8-diazabicyclo-[5.4.0]-undec-7-ene cation, and an imidazolium-based IL was produced by combining hydroxyl with imidazole. The structures of these three ILs were verified by 1H nuclear magnetic resonance and Fourier transform infrared spectroscopic analyses. The binding energies between these substances and butanol, acetone, and ethanol were calculated using the Gaussian software package based on density functional theory. The results indicated that the interaction energies between N,N-bis(2-hydroxypropyl)octan-1-aminium hydroxide imidazole ([C8DIPA][Im]) and butanol, acetone, and ethanol are greater than that with water. These ILs were examined as extractants for the separation of butanol, acetone, or ethanol with water, and the [C8DIPA][Im] exhibited the best performance. Effects of extraction time, extraction temperature, and IL proportion on extraction rate were assessed, and this IL extracted 78%, 34%, and 6% of the three components, respectively. This performance remained stable after 10 recycling trials, and [C8DIPA][Im] was also used to separate the products in an actual ABE fermentation mixture.
Background There is growing evidence that endocytosis plays a pivotal role in cancer metastasis. In this study, we first identified endocytic and metastasis-associated genes (EMGs) and then investigated the biological functions and mechanisms of EMGs. Methods Cancer stem cells (CSCs)-like characteristics were evaluated by tumour limiting dilution assays, three-dimensional (3D) spheroid cancer models. Microarray analysis was used to identify the pathways significantly regulated by mammalian Eps15 homology domain protein 1 (EHD1) knockdown. Mass spectrometry (MS) was performed to identify EHD1-interacting proteins. The function of EHD1 as a regulator of cluster of differentiation 44 (CD44) endocytic recycling and lysosomal degradation was determined by CD44 biotinylation and recycling assays. Results EHD1 was identified as a significant EMG. Knockdown of EHD1 suppressed CSCs-like characteristics, epithelial–mesenchymal transition (EMT), migration and invasion of lung adenocarcinoma (LUAD) cells by increasing Hippo kinase cascade activation. Conversely, EHD1 overexpression inhibited the Hippo pathway to promote cancer stemness and metastasis. Notably, utilising MS analysis, the CD44 protein was identified as a potential binding partner of EHD1. Furthermore, EHD1 enhanced CD44 recycling and stability. Indeed, silencing of CD44 or disruption of the EHD1/CD44 interaction enhanced Hippo pathway activity and reduced CSCs-like traits, EMT and metastasis. Interestingly, specificity protein 1 (SP1), a known downstream target gene of the Hippo-TEA-domain family members 1 (TEAD1) pathway, was found to directly bind to the EHD1 promoter region and induce its expression. Among clinical specimens, the EHD1 expression level in LUAD tissues of metastatic patients was higher than that of non-metastatic patients. Conclusions Our findings emphasise that EHD1 might be a potent anti-metastatic target and present a novel regulatory mechanism by which the EHD1/CD44/Hippo/SP1 positive feedback circuit plays pivotal roles in coupling modules of CSCs-like properties and EMT in LUAD. Targeting this loop may serve as a remedy for patients with advanced metastatic LUAD.
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