Abstract
UV and gamma irradiation mutagenesis was applied on Aspergillus fumigatus and Alternaria tenuissima in order to improve their producing ability of paclitaxel. Among the screened mutants, two stable strains (designated TXD105–GM6 and TER995–GM3) showed the maximum paclitaxel production. Paclitaxel titers of the two respective mutants were dramatically intensified to 1.22- and 1.24-fold, as compared by their respective parents. Immobilization using five different entrapment carriers of calcium alginate, agar-agar, Na-CMC, gelatin, and Arabic gum was successfully applied for production enhancement of paclitaxel by the two mutants. The immobilized cultures were superior to free-cell cultures and paclitaxel production by the immobilized mycelia was much higher than that of the immobilized spores using all the tried carriers. Moreover, calcium alginate gel beads were found the most conductive and proper entrapment carrier for maximum production of paclitaxel. The feasibility of the paclitaxel production by the immobilized mycelia as affected by incubation period, medium volume, and number of beads per flask was adopted. Under the favorable immobilization conditions, the paclitaxel titers were significantly intensified to 1.31- and 1.88-fold by the respective mutants, as compared by their free cultures. The obtained paclitaxel titers by the immobilized mycelia of the respective mutants (694.67 and 388.65 μg L−1) were found promising in terms of fungal production of paclitaxel. Hence, these findings indicate the future possibility to reduce the cost of producing paclitaxel and suggest application of the immobilization technique for the biotechnological production of paclitaxel at an industrial scale.
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References
Awan MS, Tabbasam N, Ayub N, Babar ME, Rahman M, Rana SM, Rajoka MI (2011) Gamma radiation induced mutagenesis in Aspergillus niger to enhance its microbial fermentation activity for industrial enzyme production. Mol Biol Rep 38:1367–1374. https://doi.org/10.1007/s11033-010-0239-3
Banerjee M, Chakrabarty A, Majumdar SK (1982) Immobilization of yeast cells containing β-galactosidase. Biotechnol Bioeng 24:1839–1850. https://doi.org/10.1002/bit.260240810
Bentebibel S, Moyano E, Palazo J, Rosa M, Cusido M, Bonfill M, Eibl R, Pinol MT (2005) Effects of immobilization by entrapment in alginate and scale-up on paclitaxel and baccatin III production in cell suspension cultures of Taxus baccata. Biotechnol Bioeng 89:647–655. https://doi.org/10.1002/bit.20321
Bhan N, Xu P, Koffas MAG (2013) Pathway and protein engineering approaches to produce novel and commodity small molecules. Curr Opin Biotechnol 24:1137–1143. https://doi.org/10.1016/j.copbio.2013.02.019
Cardellina JH (1991) HPLC separation of taxol and cephalomannine. J Liq Chromatogr 14:659–665. https://doi.org/10.1080/01483919108049278
Deng BW, Liu KH, Chen WQ, Ding XW, Xie XC (2009) Fusarium solani, Tax-3, a new endophytic taxol-producing fungus from Taxus chinensis. World J Microbiol Biotechnol 25:139–143. https://doi.org/10.1007/s11274-008-9876-2
Deo YM, Gaucher GM (1985) Effect of nitrogen supplementation on the longevity of antibiotic production by immobilized cells of Penicillium urticae. Appl Microbiol Biotechnol 21:220–227. https://doi.org/10.1007/BF00295126
El-Sayed ER, Ismaiel AA, Ahmed AS, Hassan IA, Karam El-Din AA (2019a) Bioprocess optimization using response surface methodology for production of the anticancer drug paclitaxel by Aspergillus fumigatus and Alternaria tenuissima: enhanced production by ultraviolet and gamma irradiation. Biocatal Agric Biotechnol 18:100996. https://doi.org/10.1016/j.bcab.2019.01.034
El-Sayed ER, Ahmed AS, Ismaiel AA (2019b) Agro-industrial byproducts for production of the immunosuppressant mycophenolic acid by Penicillium roqueforti under solid-state fermentation: enhanced production by ultraviolet and gamma irradiation. Biocatal Agric Biotechnol 18:101015. https://doi.org/10.1016/j.bcab.2019.01.053
Galazzo JL, Bailey JE (1990) Growing Saccharomyces cerevisiae in calcium alginate beads induces cell alterations which accelerate glucose conversion to ethanol. Biotechnol Bioeng 36:417–426. https://doi.org/10.1002/bit.260360413
Gils PS, Rayb D, Sahooa PK (2009) Characteristics of xanthan gum-based biodegradable superporous hydrogel. Int J Biol Macromol 45:364–371. https://doi.org/10.1016/j.ijbiomac.2009.07.007
Goksungur Y, Gunduz M, Harsa S (2005) Optimization of lactic acid production from whey by L. casei NRRL B-441 immobilized in chitosan stabilized Ca-alginate beads. J Chem Technol Biotechnol 80:1282–1290. https://doi.org/10.1002/jctb.1326
Goodman J, Walsh V (2001) The story of taxol: nature and politics in the pursuit of an anti-cancer drug. Cambridge University Press, Cambridge
Hara H, Arita T, Tachibana S, Itoh K (2008) Paclitaxel production by immobilized cell suspension cultures of Taxus cuspidate var. nana. Biotechnology 7:557–562. https://doi.org/10.3923/biotech.2008.557.562
Herdeg C, Oberhoff M, Baumbach A, Blattner A, Axel DI, Schröder S, Heinle H, Karsch KR (2000) Local paclitaxel delivery for the prevention of restenosis: biological effects and efficacy in vivo. J Am Coll Cardiol 35:1969–1976. https://doi.org/10.1016/S0735-1097(00)00614-8
Irum W, Anjum T (2012) Production enhancement of cyclosporin ‘A’ by Aspergillus terreus through mutation. Afr J Biotechnol 11:1736–1743
Ismaiel AA, Ahmed AS, El-Sayed ER (2014) Optimization of submerged fermentation conditions for immunosuppressant mycophenolic acid production by Penicillium roqueforti isolated from blue-molded cheeses: enhanced production by ultraviolet and gamma irradiation. World J Microbiol Biotechnol 30:2625–2638. https://doi.org/10.1007/s11274-014-1685-1
Ismaiel AA, Ahmed AS, El-Sayed ER (2015) Immobilization technique for enhanced production of the immunosuppressant mycophenolic acid by ultraviolet and gamma-irradiated Penicillium roqueforti. J Appl Microbiol 119:112–126. https://doi.org/10.1111/jam.12828
Ismaiel AA, Ahmed AS, Hassan IA, El-Sayed ER, Karam El-Din AA (2017) Production of paclitaxel with anticancer activity by two local fungal endophytes, Aspergillus fumigatus and Alternaria tenuissima. Appl Microbiol Biotechnol 101:5831–5846. https://doi.org/10.1007/s00253-017-8354-x
Kusari S, Hertweck C, Spiteller M (2012) Chemical ecology of endophytic fungi: origins of secondary metabolites. Chem Biol 19:792–798. https://doi.org/10.1016/j.chembiol.2012.06.004
Li Y, Zhang G, Pfeifer BA (2014) Current and emerging options for taxol production. Springer International Publishing, Cham. https://doi.org/10.1007/10_2014_292
McCabe BK, Kuek C, Gordon GLR, Phillips MW (2001) Immobilization of monocentric and polycentric types of anaerobic chytrid fungi in Ca-alginate. Enzym Microb Technol 29:144–149. https://doi.org/10.1016/S0141-0229(01)00367-2
Moayednia N, Ehsani MR, Emamdjomeh Z, Asadi MM, Mizani M, Mazaheri AF (2009) The effect of sodium alginate concentrations on viability of immobilized Lactobacillus acidophilus in fruit alginate coating during refrigerator storage. Aust J Basic Appl Sci 3:3213–3216
Nayak AK, Das B, Maji R (2012) Calcium alginate/gum Arabic beads containing glibenclamide: development and in vitro characterization. Int J Biol Macromol 51:1070–1078. https://doi.org/10.1016/j.ijbiomac.2012.08.021
Nichols NN, Dien BS, Wu YV, Cotta MA (2005) Ethanol fermentation of starch from field peas. Cereal Chem 82:554–558. https://doi.org/10.1094/CC-82-0554
Parekh S, Vinci VA, Strobel RJ (2000) Improvement of microbial strains and fermentation processes. Appl Microbiol Biotechnol 54:287–301. https://doi.org/10.1007/s002530000403
Patel RN (1998) Tour de paclitaxel: biocatalysis for semisynthesis. Ann Rev Microbiol 52:361–395. https://doi.org/10.1146/annurev.micro.52.1.361
Sallam LAR, El-Refai AMH, Hamdi AHA, El-Minofi HA, Abd-Elsalam IS (2005) Studies on the application of immobilization technique for the production of cyclosporin A by a local strain of Aspergillus terreus. J Gen Appl Microbiol 51:143–149. https://doi.org/10.2323/jgam.51.143
Scardi V (1987) Immobilization of enzymes and microbial cells in gelatin. Methods Enzymol 135:293–299. https://doi.org/10.1016/0076-6879(87)35086-4
SivaRaman H, Rao BS, Pundle AV, SivaRaman C (1982) Continuous ethanol production by yeast cells immobilized in open pore gelatin matrix. Biotechnol Lett 4:359–364. https://doi.org/10.1007/BF00135422
Strobel G, Yang X, Sears J, Kramer R, Sidhu RS, Hess WM (1996) Taxol from Pestalotiopsis microspora, an endophytic fungus of Taxus wallichiana. Microbiology 142:435–440. https://doi.org/10.1099/13500872-142-2-435
Survase SA, Annapure US, Singhal RS (2010) Gellan gum as immobilization matrix for production of cyclosporin A. J Microbiol Biotechnol 20:1086–1091. https://doi.org/10.4014/jmb.1001.01006
Terry LA, Joyce DC (2004) Elicitors of induced disease resistance in postharvest horticultural crops: a brief review. Postharvest Biol Technol 32:1–13. https://doi.org/10.1016/j.postharvbio.2003.09.016
Thacker J (1999) Repair of ionizing radiation damage in mammalian cells. Alternative pathways and their fidelity. C R Acad Sci III 322:103–108. https://doi.org/10.1016/S0764-4469(99)80030-4
West TP, Strohfus B (2001) Polysaccharide production by immobilized Aureobasidium pullulans cells in batch bioreactors. Microbiol Res 156:285–288. https://doi.org/10.1078/0944-5013-00106
Woo DD, Miao SY, Pelayo JC, Woolf AS (1994) Taxol inhibits progression of congenital polycystic kidney disease. Nature 368:750–753. https://doi.org/10.1038/368750a0
Xu Z, Yang S (2007) Production of mycophenolic acid by Penicillium brevicompactum immobilized in a rotating fibrous-bed bioreactor. Enzym Microb Technol 40:623–628. https://doi.org/10.1016/j.enzmictec.2006.05.025
Xu JF, Yin PQ, Wei XG, Su ZG (1998) Self-immobilized aggregate culture of Taxus cuspidata for improved taxol production. Biotechnol Tech 12:241–244. https://doi.org/10.1023/A:100888172
Xu F, Tao W, Cheng L, Guo L (2006) Strain improvement and optimization of the media of taxol-producing fungus Fusarium maire. Biochem Eng J 31:67–73. https://doi.org/10.1016/j.bej.2006.05.024
Xu P, Bhan N, Koffas MAG (2013) Engineering plant metabolism into microbes: from systems biology to synthetic biology. Curr Opin Biotechnol 24:291–299. https://doi.org/10.1016/j.copbio.2012.08.010
Yukimune Y, Tabata H, Higashi Y, Hara Y (1996) Methyl jasmonate-induced overproduction of paclitaxel and baccatin III in Taxus cell suspension cultures. Nat Biotechnol 14:1129–1132. https://doi.org/10.1038/nbt0996-1129
Zhang B, Maiti A, Shively S, Lakhani F, McDonald-Jones G, Bruce J, Lee EB, Xie SX, Joyce S, Li C, Toleikis PM, Lee VM, Trojanowski JQ (2005) Microtubule-binding drugs offset tau sequestration by stabilizing microtubules and reversing fast axonal transport deficits in a tauopathy model. Proc Natl Acad Sci U S A 102:227–231. https://doi.org/10.1073/pnas.0406361102
Zhang P, Zhou PP, Yu LJ (2009) An endophytic taxol-producing fungus from Taxus x media, Aspergillus candidus MD3. FEMS Microbiol Lett 293:155–159. https://doi.org/10.1111/j.15746968.2009.01481.x
Zhao K, Zhou D, Ping W, Jingping G (2004) Study on preparation and regeneration of protoplast from taxol-producing fungus Nodulisporium sylviforme. Nat Sci 2:52–59
Zhou X, Zhu H, Liu L, Lin J, Tang K (2010) A review: recent advances and future prospects of taxol-producing endophytic fungi. Appl Microbiol Biotechnol 86:1707–1717. https://doi.org/10.1007/s00253-010-2546-y
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El-Sayed, ES.R., Ahmed, A.S., Hassan, I.A. et al. Strain improvement and immobilization technique for enhanced production of the anticancer drug paclitaxel by Aspergillus fumigatus and Alternaria tenuissima. Appl Microbiol Biotechnol 103, 8923–8935 (2019). https://doi.org/10.1007/s00253-019-10129-1
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DOI: https://doi.org/10.1007/s00253-019-10129-1