Separation of cordycepin from Cordyceps militaris fermentation supernatant using preparative HPLC and evaluation of its antibacterial activity as an NAD+-dependent DNA ligase inhibitor

Abstract

Abstract. Cordycepin exhibits various bio-activities, including anticancer, antibacterial, antiviral and immune regulation activities, and is a significant focus of research. However, the preparation of high-purity cordycepin remains challenging. Also, the molecular target with which cordycepin interacts to cause an antibacterial effect remains unknown. In the present study, cordycepin was prepared by preparative high-performance liquid chromatography (prep-HPLC) and the purity obtained was 99.6%, indicating that this technique may be useful for the large-scale isolation of cordycepin in the future. The results of computational molecular docking analysis indicated that the interaction energy between cordycepin and NAD + -dependent DNA ligase (LigA) was lower than that between cordycepin and other common antibacterial targets. The highly pure cordycepin obtained by prep-HPLC demonstrated inhibitory activity against LigA from various bacteria in vitro. In conclusion, cordycepin may be useful as a broad-spectrum antibiotic targeting LigA in various bacteria. IntroductionCordycepin is an antimicrobial substance that was first isolated from the entomopathogenic fungus Cordyceps militaris by Cunningham et al in 1950 (1). Since its discovery, a number of studies have focused on this substance (2,3). Cordycepin ( Fig. 1A) with the molecular formula C 10 H 13 N 5 O 3 has been demonstrated to have various bio-activities, including anticancer (4-7), antibacterial (8), antiviral (9,10) and immune regulation activities (11) and has became a topic of interest to various research groups.However, highly pure cordycepin is challenging to obtain; various drawbacks have been identified in the purification techniques of ion resin adsorption (12,13), silica gel column chromatography (14) and supercritical extraction (15). The high price of cordycepin acts as a significant barrier to the study of its bio-activity. Preparative high performance liquid chromatography (prep-HPLC) can shorten the production cycle, has relatively low operating costs, and is simple to carry out. Therefore, it is commonly used for the purification of chemical drugs and biological drugs by pharmaceutical companies and particularly in industrial purification (16). Prep-HPLC can be used to provide purified samples on a kilogram, or even ton, scale. Also, prep-HPLC is an effective technology currently used for natural product isolation in industry (17). The present study describes a new method of obtaining high-purity cordycepin by prep-HPLC, which may be useful for the large-scale isolation of cordycepin in the future.Although studies have shown that cordycepin is a broad-spectrum antibiotic (18), studies focusing on the mechanism of the antibacterial activity and the possible targets are lacking. Molecular modeling and docking technology, which is efficient, accurate and simple to use, can reproduce the true interaction process between drug and targets that some of the current laboratory techniques are unable to detect, and is a method for predicting the mecha...

Transcriptome Analysis Reveals the Flexibility of Cordycepin Network in Cordyceps militaris Activated by L-Alanine Addition

“…By preventing excessive glutamateinduced oxidation and endoplasmic reticulum stress-associated apoptosis in hippocampal cells (Jin et al, 2014), it was also proved to have antioxidant effects (Kim et al, 2006). Besides, cordycepin was proved to have broad-spectrum antibiotic activity by inhibiting NAD + -dependent DNA ligase (Zhou et al, 2016).…”

Synergistic anticancer and antibacterial activities of cordycepin and selected natural bioactive compounds

“…The only significant synergistic antibacterial effect was produced by combination of Cor and Gen at 50 µmol/L, which resulted in much higher antibacterial effect than when the compounds were used singly. It has been previously reported that Cor is an NAD + -dependent DNA ligase inhibitor which inhibits bacterial growth [24], whereas Gen inhibits DNA and protein synthesis in Klebsiella pneumonia [25]. Therefore, Cor and Gen may mediate their antibacterial activities through different mechanisms, which manifested in a synergistic inhibitory effect on E.coli growth.…”

Anti-oxidant and anti-hyperlipidemic effects of cordycepin-rich Cordyceps militaris in a Sprague–Dawley rat model of alcohol-induced hyperlipidemia and oxidative stress

“…Cordycepin, 3-deoxyadenosine, is a derivative of the nucleoside adenosine, and it is abundantly present in Cordyceps species (Ju et al 2009;Zhou et al 2016). Since cordycepin shows chemical similarity to adenosine, it could be involved in the incorporation of RNA molecule causing a broad range of biological activities (Chen et al 2017).…”