Identification and quantification of honeybee venom constituents by multiplatform metabolomics

Abstract

Honeybee (Apis mellifera) venom (HBV) has been a subject of extensive proteomics research; however, scarce information on its metabolite composition can be found in the literature. The aim of the study was to identify and quantify the metabolites present in HBV. To gain the highest metabolite coverage, three different mass spectrometry (MS)-based methodologies were applied. In the first step, untargeted metabolomics was used, which employed high-resolution, accurate-mass Orbitrap MS. It allowed obtaining a broad overview of HBV metabolic components. Then, two targeted metabolomics approaches, which employed triple quadrupole MS, were applied to quantify metabolites in HBV samples. The untargeted metabolomics not only confirmed the presence of amines, amino acids, carbohydrates, and organic acids in HBV, but also provided information on venom components from other metabolite classes (e.g., nucleosides, alcohols, purine and pyrimidine derivatives). The combination of three MS-based metabolomics platforms facilitated the identification of 214 metabolites in HBV samples, among which 138 were quantified. The obtaining of the wide free amino acid profiles of HBV is one of the project’s achievements. Our study contributed significantly to broadening the knowledge about HBV composition and should be continued to obtain the most comprehensive metabolite profile of HBV.

Opportunities and Limitations for Untargeted Mass Spectrometry Metabolomics to Identify Biologically Active Constituents in Complex Natural Product Mixtures

“…14,45 Often, metabolomics datasets contain thousands of individually detected variables, whose signal intensities vary over a very large range, and may result from the detection of experimental artefacts. 14,46,47 Data pretreatment, filtering of chemical interferents, and model simplification tools may be critically important to enable extraction of relevant information from such datasets. 14,48 To explore this possibility in the context of natural products drug discovery, the impact of data transformation, data filtering, and model simplification, as well as their second-order interactions, were assessed using data from the ten-pool set analyzed at 100 μg/mL in both the bioassay and by the LC-MS.…”

Unraveling the venom chemistry with evidence for histamine as key regulator in the envenomation by caterpillar Automeris zaruma

“…On the other hand, free amino acids such as arginine may be present in the venom and interact with the negatively charged binding pocket. For several venomous animals such as snakes and the honey bees, it has been described that their venom contains free amino acids ( 47 , 48 ). However, more research is needed to characterize the bio-active component that activates the MRGPRX2.…”

Applications of Virtual Screening in Bioprospecting: Facts, Shifts, and Perspectives to Explore the Chemo-Structural Diversity of Natural Products

“…Recently, metabolomics and metabolic profiling approaches have explored novel taxonomic groups from the unique environment, providing opportunities for finding novel natural bioactive compounds, and some examples include bacteria (Kleigrewe et al, 2015 ; Gosse et al, 2019 ), cnidaria (Santacruz et al, 2020 ), marine sponge (Abdelhameed et al, 2020 ), insects (Klupczynska et al, 2020 ), and fungi (Oppong-Danquah et al, 2018 ). Special attention has been given to novel chemical entities that originated from marine environments due to their diverse and unique drug-like scaffolds (Shang et al, 2018 ) and physicochemical properties (Jagannathan, 2019 ) when compared with natural products of terrestrial origin, which make them a valuable source for exploration by the pharmaceutical and biotechnological industries.…”