Tryptophan decarboxylase plays an important role in ajmalicine biosynthesis in Rauvolfia verticillata
Identifieur interne : 000316 ( Istex/Checkpoint ); précédent : 000315; suivant : 000317Tryptophan decarboxylase plays an important role in ajmalicine biosynthesis in Rauvolfia verticillata
Auteurs : Wanhong Liu [République populaire de Chine] ; Rong Chen [République populaire de Chine] ; Min Chen [République populaire de Chine] ; Haoxing Zhang [République populaire de Chine] ; Meifang Peng [République populaire de Chine] ; Chunxian Yang [République populaire de Chine] ; Xingjia Ming [République populaire de Chine] ; Xiaozhong Lan [République populaire de Chine] ; Zhihua Liao [République populaire de Chine]Source :
- Planta [ 0032-0935 ] ; 2012-07-01.
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Abstract
Abstract: Tryptophan decarboxylase (TDC) converts tryptophan into tryptamine that is the indole moiety of ajmalicine. The full-length cDNA of Rauvolfia verticillata (RvTDC) was 1,772 bps that contained a 1,500-bp ORF encoding a 499-amino-acid polypeptide. Recombinant 55.5 kDa RvTDC converted tryptophan into tryptamine. The K m of RvTDC for tryptophan was 2.89 mM, higher than those reported in other TIAs-producing plants. It demonstrated that RvTDC had lower affinity to tryptophan than other plant TDCs. The K m of RvTDC was also much higher than that of strictosidine synthase and strictosidine glucosidase in Rauvolfia. This suggested that TDC might be the committed-step enzyme involved in ajmalicine biosynthesis in R. verticillata. The expression of RvTDC was slightly upregulated by MeJA; the five MEP pathway genes and SGD showed no positive response to MeJA; and STR was sharply downregulated by MeJA. MeJA-treated hairy roots produced higher level of ajmalicine (0.270 mg g−1 DW) than the EtOH control (0.183 mg g−1 DW). Highest RvTDC expression level was detected in hairy root, about respectively 11, 19, 65, and 109-fold higher than in bark, young leaf, old leaf, and root. Highest ajmalicine content was also found in hairy root (0.249 mg g−1 DW) followed by in bark (0.161 mg g−1 DW) and young leaf (0.130 mg g−1 DW), and least in root (0.014 mg g−1 DW). Generally, the expression level of RvTDC was positively consistent with the accumulation of ajmalicine. Therefore, it could be deduced that TDC might be the key enzyme involved in ajmalicine biosynthesis in Rauvolfia.
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DOI: 10.1007/s00425-012-1608-z
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Region (Ministry of Education), Laboratory of Natural Products and Metabolic Engineering, Chongqing Engineering and Technology Research Center for Sweetpotato, School of Life Sciences, Southwest University, 400715, Chongqing</wicri:regionArea><wicri:noRegion>Chongqing</wicri:noRegion></affiliation></author><author><name sortKey="Peng, Meifang" sort="Peng, Meifang" uniqKey="Peng M" first="Meifang" last="Peng">Meifang Peng</name><affiliation wicri:level="1"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Eco-environments in Three Gorges Reservoir Region (Ministry of Education), Laboratory of Natural Products and Metabolic Engineering, Chongqing Engineering and Technology Research Center for Sweetpotato, School of Life Sciences, Southwest University, 400715, Chongqing</wicri:regionArea><wicri:noRegion>Chongqing</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Chunxian" sort="Yang, Chunxian" uniqKey="Yang C" 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type="ISSN">0032-0935</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Biosynthetic pathway of alkaloids</term><term>Elicitors</term><term>Rate limiting enzyme</term><term>Terpenoid indole alkaloid</term><term>Tryptophan decarboxylase</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Tryptophan decarboxylase (TDC) converts tryptophan into tryptamine that is the indole moiety of ajmalicine. The full-length cDNA of Rauvolfia verticillata (RvTDC) was 1,772 bps that contained a 1,500-bp ORF encoding a 499-amino-acid polypeptide. Recombinant 55.5 kDa RvTDC converted tryptophan into tryptamine. The K m of RvTDC for tryptophan was 2.89 mM, higher than those reported in other TIAs-producing plants. It demonstrated that RvTDC had lower affinity to tryptophan than other plant TDCs. The K m of RvTDC was also much higher than that of strictosidine synthase and strictosidine glucosidase in Rauvolfia. This suggested that TDC might be the committed-step enzyme involved in ajmalicine biosynthesis in R. verticillata. The expression of RvTDC was slightly upregulated by MeJA; the five MEP pathway genes and SGD showed no positive response to MeJA; and STR was sharply downregulated by MeJA. MeJA-treated hairy roots produced higher level of ajmalicine (0.270 mg g−1 DW) than the EtOH control (0.183 mg g−1 DW). Highest RvTDC expression level was detected in hairy root, about respectively 11, 19, 65, and 109-fold higher than in bark, young leaf, old leaf, and root. Highest ajmalicine content was also found in hairy root (0.249 mg g−1 DW) followed by in bark (0.161 mg g−1 DW) and young leaf (0.130 mg g−1 DW), and least in root (0.014 mg g−1 DW). Generally, the expression level of RvTDC was positively consistent with the accumulation of ajmalicine. Therefore, it could be deduced that TDC might be the key enzyme involved in ajmalicine biosynthesis in Rauvolfia.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Zhejiang</li></region><settlement><li>Hangzhou</li></settlement></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Liu, Wanhong" sort="Liu, Wanhong" uniqKey="Liu W" first="Wanhong" last="Liu">Wanhong Liu</name></noRegion><name sortKey="Chen, Min" sort="Chen, Min" uniqKey="Chen M" first="Min" last="Chen">Min Chen</name><name sortKey="Chen, Rong" sort="Chen, Rong" uniqKey="Chen R" first="Rong" last="Chen">Rong Chen</name><name sortKey="Lan, Xiaozhong" sort="Lan, Xiaozhong" uniqKey="Lan X" first="Xiaozhong" last="Lan">Xiaozhong Lan</name><name sortKey="Liao, Zhihua" sort="Liao, Zhihua" uniqKey="Liao Z" first="Zhihua" last="Liao">Zhihua Liao</name><name sortKey="Liao, Zhihua" sort="Liao, Zhihua" uniqKey="Liao Z" first="Zhihua" last="Liao">Zhihua Liao</name><name sortKey="Liao, Zhihua" sort="Liao, Zhihua" uniqKey="Liao Z" first="Zhihua" last="Liao">Zhihua Liao</name><name sortKey="Ming, Xingjia" sort="Ming, Xingjia" uniqKey="Ming X" first="Xingjia" last="Ming">Xingjia Ming</name><name sortKey="Peng, Meifang" sort="Peng, Meifang" uniqKey="Peng M" first="Meifang" last="Peng">Meifang Peng</name><name sortKey="Yang, Chunxian" sort="Yang, Chunxian" uniqKey="Yang C" first="Chunxian" last="Yang">Chunxian Yang</name><name sortKey="Zhang, Haoxing" sort="Zhang, Haoxing" uniqKey="Zhang H" first="Haoxing" last="Zhang">Haoxing Zhang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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