L-tryptophan (Trp) is widely used in food and feed enforcement to play an important role in biological processes. Various metabolites of Trp perform its potent function. The indole pyruvate pathway is one of the main pathways of Trp metabolism in the gut microbiota, providing numerous indole-derivatives, which can modulate intestinal homeostasis and mucosal immunity by activating the aryl hydrocarbon receptor (AHR) signaling pathway. In this study, we constructed an IL4I1-overexpressed 293T cell line and found that IL4I1 can catalyze Trp to produce indole-3-acetic acid (IAA) and indole-3-carboxaldehyde (IAld). Moreover, both IAA and IAld are accumulated in dendritic cells (DCs) and can stimulate the expression of CYP1A1. Our results demonstrate the existence of the indole pyruvate pathway in host cells with IL4I1 as the key enzyme. The IL4I1-mediated Trp metabolism implies the role of dietary impact on immunity.

The novelty of this method is that the commonly used fluorinated ion-pairing reagent was not used in the mobile phase in our analysis, greatly reducing the contamination of the ESI source in negative mode. Moreover, the four gentamicin components were clearly separated via chromatographic separation.

The macrolide regulatory protein MphR(A) has been widely studied and used in various aspects such as metabolism monitoring, exogenous gene expression, and in vivo and in vitro macrolide antibiotic screening. Another macrolide regulatory protein, MphR(E), has rarely been reported. In this study, in vitro ELISA-type systems were established for MphR(A) and MphR(E) to study their correlation. The reactivity of 14 macrolide antibiotics and pseudo-macrolide antibiotics was tested in the systems. The results indicated that the ligand identification spectra of MphR(A) and MphR(E) were basically consistent. The binding characteristics of MphR(A) and MphR(E) with three corresponding promoter DNA sequences were preliminarily studied. According to the ELISA-type analysis results, MphR(A) and MphR(E) have consistent DNA binding properties, which bind to A-DNA/B-DNA more easily than to C-DNA. This study has confirmed that MphR(E) can bind to the promoter DNA sequences mrx(E) and mph(E) in plasmid pRSB111, and different DNAs can affect the sensitivity of the in vitro detection systems.