Chemiluminescence Detection of Hydrogen Sulfide Release by β-Lactamase-Catalyzed β-Lactam Biodegradation: Unprecedented Pathway for Monitoring β-Lactam Antibiotic Bacterial Resistance

Abstract

β-Lactamase positive bacteria represent a growing threat to human health because of their resistance to commonly used antibiotics. Therefore, development of new diagnostic methods for identification of β-lactamase positive bacteria is of high importance for monitoring the spread of antibiotic-resistant bacteria. Here, we report the discovery of a new biodegradation metabolite (H 2 S), generated through β-lactamase-catalyzed hydrolysis of β-lactam antibiotics. This discovery directed us to develop a distinct molecular technique for monitoring bacterial antibiotic resistance. The technique is based on a highly efficient chemiluminescence probe, designed for detection of the metabolite, hydrogen sulfide, that is released upon biodegradation of β-lactam by β-lactamases. Such an assay can directly indicate if antibiotic bacterial resistance exists for a certain examined β-lactam. The assay was successfully demonstrated for five different β-lactam antibiotics and eight β-lactam resistant bacterial strains. Importantly, in a functional bacterial assay, our chemiluminescence probe was able to clearly distinguish between a β-lactam resistant bacterial strain and a sensitive one. As far as we know, there is no previous documentation for such a biodegradation pathway of β-lactam antibiotics. Bearing in mind the data obtained in this study, we propose that hydrogen sulfide should be considered as an emerging β-lactam metabolite for detection of bacterial resistance.

Seeking Illumination: The Path to Chemiluminescent 1,2-Dioxetanes for Quantitative Measurements and In Vivo Imaging

“…In comparison to the CHS probes series, Simsek Turan and Somzen, and Levinn and Pluth have reported probes for hydrogen sulfide using dinitrophenyl and azide triggers, although these studies were limited to in vitro experiments. More recently, Gholap and co-workers reported a series of hydrogen sulfide 1,2-dioxetane probes with various triggers and used these to study hydrogen sulfide production from β-Lactam antibiotic degradation. Amongst these studies, only the CHS series has been used for in vivo imaging of hydrogen sulfide.…”

Enzyme‐Activated, Chemiluminescent Siderophore‐Dioxetane Probes Enable the Selective and Highly Sensitive Detection of Bacterial Pathogens

“…This resulted in activatable probes that are brightly emissive in aqueous environments and applicable in biological systems [13–15] . Lately, we functionalized dioxetanes for the detection of Mycobacterium tuberculosis as well as for bacterial carbapenemases and β‐lactamases [16–18] …”

“…[13][14][15] Lately, we functionalized dioxetanes for the detection of Mycobacterium tuberculosis as well as for bacterial carbapenemases and β-lactamases. [16][17][18] However, a broad detection of both Gram-negative and positive bacteria that does not rely on the secretion of resistance enzymes like β-lactamases remained elusive so far. One cause for this is the hampered intracellular accumulation and activation of such probes, impeded by impervious bacterial cell walls.…”

Enzymaktivierte, chemilumineszente Siderophor‐Dioxetan‐Sonden ermöglichen den selektiven und hochempfindlichen Nachweis von bakteriellen Krankheitserregern

“…Durch die Modifikation wurden enzymatisch aktivierbare Sonden erhalten, welche auch in wässriger Umgebung hell emittieren und somit in biologischen Systemen anwendbar sind [13–15] . Kürzlich haben wir Dioxetane für den Nachweis von Mykobakterium tuberculosis sowie für bakterielle Carbapenemasen und β‐Laktamasen funktionalisiert [16–18] …”

“…[13][14][15] Kürzlich haben wir Dioxetane für den Nachweis von Mykobakterium tuberculosis sowie für bakterielle Carbapenemasen und β-Laktamasen funktionalisiert. [16][17][18] Der Nachweis von sowohl gram-negativen als auch gram-positiven Bakterien, welcher nicht auf der Sekretion von Resistenzenzymen wie β-Laktamasen beruht, ist jedoch bisher nicht gelungen. Eine Ursache dafür ist die erschwerte intrazelluläre Anreicherung und Aktivierung solcher Sonden, weil die schwer durchlässigen bakteriellen Zellwände eine effektive Barriere darstellen.…”