-Carbamoylputrescine Amidohydrolase of , a Dominant Species of the Human Gut Microbiota

Overview

Journal Biomedicines

Specialty Biomedical Engineering

Date 2023 May 16

PMID 37189741

Authors

Hiromi Shimokawa

Mikiyasu Sakanaka

Yuki Fujisawa

Hirokazu Ohta

Yuta Sugiyama

Shin Kurihara

Affiliations

Soon will be listed here.

Abstract

Polyamines are bioactive amines that play a variety of roles, such as promoting cell proliferation and protein synthesis, and the intestinal lumen contains up to several mM polyamines derived from the gut microbiota. In the present study, we conducted genetic and biochemical analyses of the polyamine biosynthetic enzyme -carbamoylputrescine amidohydrolase (NCPAH) that converts -carbamoylputrescine to putrescine, a precursor of spermidine in which is one of the most dominant species in the human gut microbiota. First, gene deletion and complemented strains were generated, and the intracellular polyamines of these strains cultured in a polyamine-free minimal medium were analyzed using high-performance liquid chromatography. The results showed that spermidine detected in the parental and complemented strains was depleted in the gene deletion strain. Next, purified NCPAH-(His) was analyzed for enzymatic activity and found to be capable of converting -carbamoylputrescine to putrescine, with a Michaelis constant () and turnover number () of 730 µM and 0.8 s, respectively. Furthermore, the NCPAH activity was strongly (>80%) inhibited by agmatine and spermidine, and moderately (≈50%) inhibited by putrescine. This feedback inhibition regulates the reaction catalyzed by NCPAH and may play a role in intracellular polyamine homeostasis in .

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