Taxoids profiling of suspension Taxus chinensis var. mairei cells in response to shear stress |
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| Institution: | 1. Key Laboratory of Systems Bioengineering, Ministry of Education and Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, PR China;2. Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR China;3. Tianjin High-standard Quality Testing Laboratory, Huada Road, 17th Kilometric Marker of Jinjing Speedway, Xiqing District, Tianjin, PR China;1. Institute of Environmental Research (INFU), Department of Chemistry and Chemical Biology, Chair of Environmental Chemistry and Analytical Chemistry, TU Dortmund, Otto-Hahn-Str. 6, D-44221 Dortmund, Germany;2. Department of Biochemistry, Indian Institute of Science (IISc), Bangalore 560012, Karnataka, India;1. Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032, West Bengal, India;2. Biophysics and Structural Genomics Division, Saha Institute of Nuclear Physics, Kolkata 700064, West Bengal, India;3. Division of Microbiology, Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032, West Bengal, India;4. Department of Environmental Science, University of Calcutta, Kolkata 700019, India;1. Shanghai Municipality Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China;2. Laboratory of Regenerative Medicine, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China;3. Department of Obstetrics and Gynecology, Shanghai Jiao Tong University Affiliated First People''s Hospital, Shanghai 200080, China |
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| Abstract: | To develop an optimal bioprocess for paclitaxel (Taxols) supply, taxoid biosynthetic pathway regulation must be better understood. The main taxoid metabolites (paclitaxel, baccatin III, taxol C, etc.) in Taxus cell culture showed great difference under shear stress. However, the regulating mechanism of taxoids metabolism under shear stress remained elusive. Here an efficient metabolic profiling approach combined with multivariate analysis was employed to profile taxoids changes of Taxus cells under laminar shear stress. A total of 21 taxoids were identified and quantified by ultra-performance liquid chromatography coupled with Q-TOF mass spectrometry. The result showed the contents of paclitaxel and baccatin III were reduced by shear stress, indicating the inhibitory effect of shear stress on paclitaxel biosynthesis. The levels of other taxoids uninvolved in paclitaxel biosynthesis were decreased except several metabolites. Further analysis of mapping measured taxoids concentrations onto paclitaxel biosynthesis pathway illustrating proposed intermediates and “off-pathway” metabolites revealed shear stress might disrupt the appropriate cyclization process of geranylgeranyl-pyrophosphate, aggravate the inappropriate order of hydroxylations and acylations, and not be good for functional group oxetane formation. These findings revealed the possible mechanism for shear stress limiting paclitaxel production and might have important biotechnological applications to increase the yields of paclitaxel and relevant precursors. |
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| Keywords: | Taxoids Plant cell culture Hydrodynamic shear stress Bioreactors Large scale cultivation Physiology |
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