Abstract
Paclitaxel is the top-selling chemotherapeutic drug used for the treatment of lung, ovarian and breast cancer as well as Kaposi’s sarcoma. Cell suspension culture (CSC) of Corylus avellana has been addressed as a promising alternative for producing paclitaxel. In this study, endophytic fungus strain YEF33 was isolated from Taxus baccata and identified as Coniothyrium palmarum. The effects of the elicitors derived from this fungus including cell extract, culture filtrate and cell wall (CW) and also chitin, alone or in combination with Methyl-β-Cyclodextrin (MBCD), on paclitaxel biosynthesis in C. avellana CSC were assayed for the first time. CW of C. palmarum was the most efficient fungal elicitor for paclitaxel biosynthesis in C. avellana CSC. The results revealed that MBCD affected paclitaxel biosynthesis differently depending on fungal elicitor type and vice versa. MBCD, either alone or in combination with fungal elicitors, induced a high secretion of paclitaxel, suggesting the decrement of toxicity and retro-inhibition processes of paclitaxel for cells. The joint effects of C. palmarum CW (2.5% (v/v) on 17th day) and 50 mM MBCD synergistically enhanced paclitaxel biosynthesis (402.4 µg l−1; 5.8-fold), 78.6% of which (316.5 µg l−1) were secreted into culture medium, a level 146% higher than that in control.
Highlights
IntroductionHigh-yielding in vitro culture setup is essential for producing secondary metabolites through plant cell culture[19]
Paclitaxel, the most effective chemotherapy agent against lung, ovarian and breast cancer, and Kaposi’s sarcoma[1], was originally extracted from Taxus brevifolia bark in 19672 and its structure was published in 19713, and it was joined the drug development program of National Cancer Institute (NCI)[4]
Our results show that the influences of fungal elicitors on paclitaxel biosynthesis are affected by fungal elicitor concentration levels and its adding time to cell culture
Summary
High-yielding in vitro culture setup is essential for producing secondary metabolites through plant cell culture[19]. Screening the efficient elicitors for stimulating the biosynthesis of secondary metabolite in a plant cell culture system is vital[24]. Fungal elicitors because of their high effectiveness and low cytotoxicity are mainly used for inducing the biosynthesis of secondary metabolites in plant cell cultures[25]. Chitin is one of fungal MAMPs29 and induces the biosynthesis of different secondary metabolites in plant cell cultures[30,31,32,33]. No data is available respecting the effects of chitin, fungal cell wall and comparing the efficiency of different fungal elicitors on paclitaxel content enhancement in C. avellana CSC. The combined use of biotic and abiotic elicitors in Taxus[37] and Corylus avellana[38] CSCs has been shown to highly enhance the biosynthesis of paclitaxel
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