Enhanced production of tropane alkaloids in <Emphasis Type="Italic">Scopolia parviflora</Emphasis> by introducing the <Emphasis Type="Italic">PMT</Emphasis> (putrescine <Emphasis Type="Italic">N</Emphasis>-methyltransferase) gene |
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Authors: | Ok-Sun?Lee Young-Min?Kang Hee-Young?Jung Ji-Yun?Min Seung-Mi?Kang Chandrakant?S?Karigar D?Theertha?Prasad Jung-Dong?Bahk Email author" target="_blank">Myung-Suk?ChoiEmail author |
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Institution: | (1) Division of Applied Life Science, Gyengsang National University, 660701 Jinju, Korea;(2) Division of Forest Science, Gyeongsang National University, 660701 Jinju, Korea;(3) Division of Biochemistry, Department of Chemistry, Central College Campus, Bangalore University, 560001 Bangalore, India;(4) Department of Biotechnology, University of Agricultural Science, GKVK, 560065 Bangalore, India;(5) Environmental Biotechnology Research Center, Gyeongsang National University, 660701 Jinju, Korea |
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Abstract: | Summary In wild-type Scopolia parvilfora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropanane
alkaloid levels were the highest in the root (0.9 mg g−1 on a dry weight basis), followed by the stem and then the leaves. We metabolically engineered S. parviflora by introducing the tobacco pmt gene into its genome by a binary vector system that employs disarmed Agrobacterium rhizogenes. The kanamycin-resistant hairy root lines were shown to bear the pmt gene and to overexpress its mRNA and protein product by at least two-fold, as determined by polymerase chain reaction (PCR)
and Northern and Western blottings, respectively. The transgenic lines also showed higher PMT activity and were morphologically
aberrant in terms of slower growth and the production of lateral roots. The overexpression of pmt markedly elevated the scopolamine and hyoscyamine levels in the transgenic lines that showed the highest pmt mRNA and PMT protein levels. Thus, overexpression of the upstream regulator of the tropane alkaloid pathway enhanced the
biosynthesis of the final product. These observations may be useful in establishing root culture systems that generate large
yields of tropane alkaloids.
These authors contributed equally to this paper (co-first authors). |
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Keywords: | hairy root metabolic engineering putrescine N-methyltransferase Scopolia parviflora tropane alkaloids |
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