Development of an enzyme linked immunosorbent assay for detection of cyathane diterpenoids

Abstract

BackgroundSo-called cyathane type diterpenoids are produced as secondary metabolites by basidiomycetes. Based on their antibacterial, fungicidal, and cytotoxic properties, cyathane type terpenoids represent interesting target compounds in fungal biotechnology.ResultsAn indirect competitive enzyme linked immunosorbent assay has been developed for detection of cyathane type diterpenoids. Rabbit polyclonal antibodies were raised against a mixture of striatal A and B conjugated to bovine serum albumin. The conditions for direct attachment of the hapten striatal B to a solid phase by passive adsorption were optimized. The cross reactivities of the striatals A, C and D, of the striatins A and B, and of the erinacines C and P to striatal B were determined. The validation study showed that the ELISA was precise and sensitive. The average IC50 of striatal B was 36.0 ng mL−1 with an inter-assay coefficient of variation (CV) of 13.2% (n = 5). Recoveries from striatal B spiked samples in the assay were in the range of 97.3 – 125.9%. A good correlation between the striatal B concentration measured by the ELISA and by HPLC-DAD (y = 1.1122× – 0.1585, R2 = 0.9942) was obtained from linear regression analysis. The suitability of the ELISA for detection of cyathane type diterpenoids in submerged cultures and fruiting bodies of H. erinaceus was studied. It showed cross reactivity with supernatants from submerged cultures and extracts thereof, but did not show cross reactivity with extracts from fruiting bodies.ConclusionsThe developed method is appropriate for qualitative and quantitative detection of cyathane diterpenoids in complex mixtures. Due to its high sensitivity and specificity, it represents an ideal screening method for discovering new cyathane diterpenoids and new potential producers of them.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-014-0098-4) contains supplementary material, which is available to authorized users.

Production of cyathane type secondary metabolites by submerged cultures of Hericium erinaceus and evaluation of their antibacterial activity by direct bioautography

“…Reference substances of erinacine C and erinacine P were isolated from extracts of H. erinaceus cultures by preparative HPLC and identified by MS and NMR analyses as described previously [ 25 ].…”

A novel immunoassay for quantitative drug abuse screening in serum

“…The anti-benzoylecgonine antibody could be used in further experiments. It is a very well-known effect of antibodies raised against small molecules to show cross-reactivities against chemically related compounds(Castaneto et al, 2015;Shen et al, 2014) like it was observed for the antimethamphetamine antibody (figure 1a…”

Secondary Metabolites of Bird’s Nest Fungi: Chemical Structures and Biological Activities

“…Subsequently, these compounds were found to hydrolyze and generate three new cyathane-xylosides, striatals A–C ( 92 – 94 ), containing ketone and aldehyde groups . Striatal D ( 95 ) was first isolated from Gerronema fibula and later from C. striatus , together with 92 – 94 . Six additional highly oxygenated polycyclic cyathane diterpene xylatoids, striatoids A–F ( 96 – 101 ), were isolated from C. striatus , with 97 and 98 exhibiting an unusual 15, 4′-ether ring system .…”

“…40 Striatal D (95) was first isolated from Gerronema f ibula 41 and later from C. striatus, together with 92−94. 42 Six additional highly oxygenated polycyclic cyathane diterpene xylatoids, striatoids A−F (96− 101), were isolated from C. striatus, with 97 and 98 exhibiting an unusual 15, 4′-ether ring system. 43 Cyathinins A−E (102− 106) and 10-hydroxyerinacine S (107), 44 which is a hydroxylation product of C-10 of erinacine S (107a) from H. erinaceum, were discovered in a new Cyathus species, C. subglobisporus BCC44381.…”