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

BMC Biotechnology

Tian Shen²  Lena M Hof²  Heike Hausmann¹  Marc Stadler³  Holger Zorn² 

[1] Justus Liebig University Giessen, Institute of Organic Chemistry, Heinrich-Buff-Ring 58, Giessen 35392, Germany

[2] Justus Liebig University Giessen, Institute of Food Chemistry and Food Biotechnology, Heinrich-Buff-Ring 58, Giessen 35392, Germany

[3] Department Microbial Drugs, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, Braunschweig 38124, Germany

关键词: Erinacine;    Striatal;    Hericium erinaceus;    ELISA;    Cyathane diterpenoids;   

Others  :  1084421
DOI  :  10.1186/s12896-014-0098-4

 received in 2014-06-02, accepted in 2014-11-05,  发布年份 2014

【 摘 要 】

Background

So-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.

Results

An 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.

Conclusions

The 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.

【 授权许可】

   
2014 Shen et al.; licensee BioMed Central Ltd.

【 图 表 】

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