ﺑﻬﯿﻨﻪﺳﺎزي ﺗﻮﻟﯿﺪ ﺗﺒﺎﯾﯿﻦ از رﯾﺸﻪﻫﺎي ﻧﺎﺑﺠﺎي ﺧﺸﺨﺎش ﮐﺒﯿﺮ Papaver bracteatum Lindle.)( ﺑﺎ ﺗﻐﯿﯿﺮ ﺣﺠﻢ ﻫﻮادﻫﯽ و دﻣﺎي ﺑﯿﻮراﮐﺘﻮر

Optimization of Thebaine Production using Adventitious Roots of Papaver bracteatum Lindle. by Alteration of Aeration Volume and Temperature in Bioreactor

ﻣﻘﺪﻣﻪ: اﻣﺮوزه ﺑﻬﯿﻨﻪﺳﺎزي ﺷﺮاﯾﻂ ﮐﺸﺖ در ﺑﯿﻮراﮐﺘﻮرﻫﺎ ﺑﻪ ﻋﻨﻮان ﯾﮏ راﻫﮑﺎر ﻣﻬﻢ ﺑﺮاي ﺗﻮﻟﯿﺪ ﻣﺘﺎﺑﻮﻟﯿﺖﻫﺎي ﺛﺎﻧﻮﯾﻪ از ﺟﻤﻠﻪ ﺗﺒﺎﺋﯿﻦ ﺑﺴﯿﺎر ﻣﻮرد ﺗﻮﺟﻪ اﺳﺖ. ﺗﺒﺎﺋﯿﻦ، آﻟﮑﺎﻟﻮﺋﯿﺪ ﻏﺎﻟﺐ در ﮔﯿﺎه ﺧﺸﺨﺎش اﯾﺮاﻧﯽ اﺳﺖ ﮐﻪ ﺑﻪ ﻋﻨﻮان ﻣﺎده اوﻟﯿﻪ ﺟﻬﺖ ﺳﻨﺘﺰ ﺗﺮﮐﯿﺒﺎت ﺿﺪ درد ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﻣﯽﮔﯿﺮد. ﻫﺪف: ﺑﻬﯿﻨﻪﺳﺎزي ﺷﺮاﯾﻂ ﮐﺸﺖ رﯾﺸﻪ ﻧﺎﺑﺠﺎ ﺧﺸﺨﺎش ﮐﺒﯿﺮ در ﺑﯿﻮراﮐﺘﻮر در راﺳﺘﺎي اﻓﺰاﯾﺶ ﺗﻮﻟﯿﺪ ﺗﺒﺎﯾﯿﻦ. روش ﺑﺮرﺳﯽ: در اﯾﻦ آزﻣﺎﯾﺶ، رﯾﺸﻪﻫﺎي ﻧﺎﺑﺠﺎ از رﯾﺰﻧﻤﻮﻧﻪ ﺳﺎﻗﻪ ﮔﯿﺎه ﺑﻪ دﺳﺖ آﻣﺪه و ﺳﭙﺲ در ﺑﯿﻮراﮐﺘﻮر ﺳﺘﻮﻧﯽ ﺣﺒﺎبدار ﮐﺸﺖ ﺷﺪﻧﺪ. اﯾﻦ آزﻣﺎﯾﺶ ﺑﻪ ﺻﻮرت ﻓﺎﮐﺘﻮرﯾﻞ در ﻗﺎﻟﺐ ﻃﺮح ﮐﺎﻣﻼً ﺗﺼﺎدﻓﯽ اﻧﺠﺎم ﺷﺪ. ﻓﺎﮐﺘﻮر دﻣﺎي ﺑﯿﻮراﮐﺘﻮر در 4 ﺳﻄﺢ ﺷﺎﻣﻞ 20 ،14، 26 و 32 درﺟﻪ ﺳﺎﻧﺘﯽﮔﺮاد و ﻫﻤﭽﻨﯿﻦ ﺣﺠﻢ ﻫﻮادﻫﯽ ﺑﯿﻮراﮐﺘﻮر در 4 ﺳﻄﺢ )0/2 ،0/1 ،0/05 و vvm 0/4( اﻋﻤﺎل ﺷﺪﻧﺪ. ﻧﺘﺎﯾﺞ: ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﮐﻪ دﻣﺎ و ﺣﺠﻢ ﻫﻮادﻫﯽ ﺑﯿﻮراﮐﺘﻮر ﺗﺄﺛﯿﺮ ﻣﻌﻨﯽداري ﺑﺮ وزن ﺧﺸﮏ رﯾﺸﻪ و ﻣﯿﺰان ﺗﺒﺎﯾﯿﻦ آن داﺷﺘﻨﺪ. ﺑﯿﺸﺘﺮﯾﻦ وزن ﺧﺸﮏ رﯾﺸﻪ ﻧﺎﺑﺠﺎ در ﻫﻮادﻫﯽ vvm 0/2 و در دﻣﺎي 26 درﺟﻪ ﺳﺎﻧﺘﯽﮔﺮاد ﺑﻪ دﺳﺖ آﻣﺪ. ﺑﯿﺸﺘﺮﯾﻦ ﻣﯿﺰان ﺗﺒﺎﯾﯿﻦ رﯾﺸﻪ ﻧﺎﺑﺠﺎ در ﺣﺠﻢ ﻫﻮادﻫﯽ vvm 0/2 و در دﻣﺎي 26 درﺟﻪ ﺳﺎﻧﺘﯽﮔﺮاد ﺣﺎﺻﻞ ﺷﺪ. ﻧﺘﯿﺠﻪﮔﯿﺮي: ﺣﺠﻢ ﻫﻮادﻫﯽ و دﻣﺎ دو ﻓﺎﮐﺘﻮر ﻣﻬﻢ در ﺗﻮﻟﯿﺪ اﻧﺒﻮه زﯾﺴﺖ ﺗﻮده و ﺗﺒﺎﯾﯿﻦ از رﯾﺸﻪ ﻧﺎﺑﺠﺎ ﺧﺸﺨﺎش ﮐﺒﯿﺮ در ﺑﯿﻮراﮐﺘﻮر ﻣﯽﺑﺎﺷﺪ.

Background: Nowadays, the optimization of the culture conditions in bioreactors is considered as an approach to produce secondary metabolites such as thebaine. Thebaine is the dominant alkaloid in Iranian poppy that is used as a precursor for the synthesis of analgesic compounds. Objective: Optimization of culture conditions of Papaver bracteatum adventitious roots in bioreactor for the scale-up thebaine production. Methods: In this study, adventitious roots was induced from the stem explants and then cultured in a bubble column bioreactor. The research was conducted as a factorial experiment based on randomized complete design. The bioreactor temperatures were 14, 20, 26 and 32 °C, as well as aeration volumes were 0.05, 0.1, 0.2 and 0.4 vvm. Results: The results showed that the temperature and aeration volume had significant effect on root fresh weight and thebaine content. The highest root dry weight was related to aeration volume of 0.2 vvm and temperature of 26°C. The maximum content of thebaine was observed in aeration volume of 0.2 vvm and temperature of 26°C. Conclusion: According to the results, aeration volume and temperature were two important factors for large-scale production of Papaver bracteatum biomass and thebaine in bioreactor conditions.