A cellulase-supported two-phase in situ system for enhanced biosynthesis of paclitaxel in Taxus × media hairy roots

• Published in: Acta physiologiae plantarum Vol. 40; no. 11; pp. 1 - 11
• Main Authors: Sykłowska-Baranek, K., Szala, K., Pilarek, M., Orzechowski, R., Pietrosiuk, A.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.11.2018; Springer Nature B.V
• Subjects: Aeration; Agriculture; Biomedical and Life Sciences; Biosynthesis; Cell culture; Cellulase; Cellulolytic enzymes; Dosage; Enzymes; Hairy root; Life Sciences; Liquid phases; Methyl jasmonate; Original Article; Paclitaxel; Perfluorodecalin; Phenylalanine; Plant Anatomy/Development; Plant Biochemistry; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Sodium; Sodium nitroprusside; Sucrose; Sugar; System effectiveness; Taxus media; Elicitation; Perfluorodecalin; Hairy roots; In vitro paclitaxel release
• ISSN: 0137-5881; 1861-1664
• DOI: 10.1007/s11738-018-2777-6

The effectiveness of two liquid-phase culture systems, in situ supported with perfluorodecalin (PFD), and elicited with methyl jasmonate (100 µM) and sodium nitroprusside (10 µM) (spiked with l -phenylalanine (100 µM) and sucrose (30 g/l) feeding), and additionally combined with cellulolytic enzyme application (Cellulase or Viscozyme ® L at doses 0.01%, 0.1%, 0.5%  or 1%), was investigated on the enhancement of paclitaxel release in Taxus × media harbouring taxadiene synthase transgene hairy root cultures. Neither elicitation nor in situ application of PFD significantly had an effect on root growth until enzyme addition; however, the hairy root response to the culture conditions varied depending on enzyme type and dosage. The highest paclitaxel total yield (intracellular + extracellular) was determined in the one-phase elicited culture systems without enzymes and amounted to 264.2 µg/flask (1 434.9 ± 516.6 µg/g DW) and 29.6 µg/flask in root biomass and medium phase, respectively. Although the application of cellulolytic enzymes did not enhance the total paclitaxel production, they intensified paclitaxel release in a dose-depending manner. Among two examined cellulolytic enzyme treatments, Viscozyme ® L addition caused the highest paclitaxel release up to 59% of its total content when used at a concentration of 0.01%. Whereas in two-phase in situ systems, the combination of Viscozyme ® L at dosage of 0.5% and PFD-aerated, resulted in the highest extracellular paclitaxel concentration up to 20%.