Fungal Cell Wall and Methyl-β–Cyclodextrin Synergistically Enhance Paclitaxel Biosynthesis and Secretion in Corylus avellana Cell Suspension Culture

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 Yef 33 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 retroinhibition processes of paclitaxel for cells. The joint effects of C. palmarum CW (2.5% (v/v) on 17 th 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. Paclitaxel, the most effective chemotherapy agent against lung, ovarian and breast cancer, and also Kaposi's sarcoma 1 , was originally extracted from Taxus brevifolia bark in 1967 2 and its structure was published in 1971 3 , and then it was joined the drug development program of National Cancer Institute (NCI) 4. Since the bark harvest is mortal for the trees, T. brevifolia was set on the endangered species list 4,5. Plant cell suspension culture (CSC) is a hopeful and nature-friendly system to mass production of paclitaxel 6-8. The worldwide demand for paclitaxel is rising at a high speed and its biosynthesis via Taxus cell factories is inadequate to handle the growing need of this medicine, mostly because of Taxus recalcitrant manner under in vitro conditions 6,7,9,10. Thus, finding the alternative sources of this valuable secondary metabolite is prompted. Corylus avellana, common hazel, has likewise been reported as a paclitaxel-producing species among angiosperms 6,7,10-15. The major superiority of producing paclitaxel by C. acellana cell culture is that in vitro culture of C. avellana is more facile than that of Taxus 6,7,9,12,16,17. In vitro culture of C. avellana has been reported as a hopeful and inexpensive method for producing paclitaxel 6,7,10,12,18. High-yielding in vitro culture setup is essential for producing secondary metabolites through plant cell culture 19. Bioactive compounds are usually fluctuated quantitatively/qualitatively under different conditions either in vivo or in vitro 6,7,10,12,20-23. Even engineered plant cells for overexpressing key genes still need using the elicitors for mass-biosynthesis of relevant secondary metabolite. Thus, screening the efficient elicitors for s...

Mathematical Modeling of Growth and Paclitaxel Biosynthesis in Corylus avellana Cell Culture Responding to Fungal Elicitors Using Multilayer Perceptron-Genetic Algorithm

“…Based on our previous studies (Salehi et al, 2019b;Salehi et al, 2019c;Farhadi et al, 2020), three concentrations [2.5, 5, and 10% (v/v)] of fungal elicitors "CE:CF (100:0, 75:25, 50:50, 25:75, 0:100 v/v)" and also mid (day 13) and late (day 17) log phase of C. avellana cell cultures were selected for adding fungal elicitors. Control received an equal volume of water (for CE)/ PDB (for CF).…”

“…Previous studies (Salehi et al, 2019b;Salehi et al, 2019c;Farhadi et al, 2020;Salehi et al, 2020) demonstrated the positive influences of cell extract (CE) and culture filtrate (CF) of endophytic fungi on paclitaxel biosynthesis in cell suspension culture (CSC) of C. avellana. Fungal elicitor type, concentration and adding time, and also exposure time of cell culture (CSC harvesting time) should be optimized to achieve the maximum biosynthesis of paclitaxel in C. avellana CSC (Salehi et al, 2019b;Salehi et al, 2019c;Farhadi et al, 2020;Salehi et al, 2020). Precise analysis of the effects of these factors and their optimal selection would pave the way for the commercialization of bioprocessing C. avellana cells toward paclitaxel mass production.…”

Whole fungal elicitors boost paclitaxel biosynthesis induction in Corylus avellana cell culture

“…Since Taxus trees are slow-growing and harvesting the bark is destructive [ 5 ], thus the continuous harvest of native plant bark for commercial production of paclitaxel was untenable. Nowadays, plant cell factories offer a promising and environment-friendly approach for large-scale paclitaxel production [ 6 – 10 ]. The rising demand for paclitaxel, and Taxus recalcitrant behavior under in vitro conditions have caused extensive efforts toward finding alternatives for producing this invaluable secondary metabolite.…”

“…avellana is a dicotyledonous plant, possibly C . avellana response to genetic manipulation by Agrobacterium is more promising than that of Taxus [ 7 – 10 ]. Obtaining high-producing in vitro cultures is a key step for producing secondary metabolites through plant cell culture (PCC) [ 13 ].…”

“…Obtaining high-producing in vitro cultures is a key step for producing secondary metabolites through plant cell culture (PCC) [ 13 ]. The various factors affect the biosynthesis of bioactive compounds in plants [ 7 – 10 , 12 , 14 – 17 ]. Amongst the available techniques for enhancing secondary metabolites biosynthesis in PCC, the elicitation is likely the most effective one for dramatic increment in yield [ 18 , 19 ].…”