TY - JOUR
T1 - Strictosidine-related enzymes involved in the alkaloid biosynthesis of Uncaria tomentosa root cultures grown under oxidative stress
AU - Vera-Reyes, Ileana
AU - Huerta-Heredia, Ariana A.
AU - Ponce-Noyola, Teresa
AU - Flores-Sanchez, Isvett Josefina
AU - Esparza-García, Fernando
AU - Cerda-García-Rojas, Carlos M.
AU - Trejo-Tapia, Gabriela
AU - Ramos-Valdivia, Ana C.
PY - 2013/5
Y1 - 2013/5
N2 - The activity and gene expression of strictosidine-related enzymes in Uncaria tomentosa root cultures exposed to oxidative stress were studied. Elicitation with 0.2 mM hydrogen peroxide (H2O2) or a combination of 0.8 mM buthionine sulfoximine and 0.2 mM jasmonic acid (BSO-JA) increased peroxidase activities by twofold at Day 8 and glutathione reductase by 1.4-fold at Day 5 in H2O2 elicited cultures respect to the control. Production of monoterpenoid oxindole alkaloids (MOA), 3α-dihydrocadambine, and dolichantoside was stimulated after H2O2 elicitation, reaching levels of 886.4±23.6, 847.7±25.4, and 87.5±7.2 μg/g DW, at Day 8 which were 1.7-, 2.1-, and 2.3-fold higher relative to control. BSO-JA elicited cultures produced about twice alkaloids than H2O2-treated cultures, following a biphasic pattern with maxima at 0.5 and 8 days. Alkaloid production was preceded by increase in strictosidine synthase (STR) and strictosidine glucosidase (SGD) activities. After elicitation with H2O2 or BSO-JA, the STR activity (pKat/mg protein) increased by 1.9-fold (93.8±17.8 at 24 h) or 2.5-fold (102.4±2.2 at 6 h) and the SGD activity (pKat/mg protein) by 2.8-fold (245.2±14.4 at 6 h) or 4.2-fold (421.2±1.8 at 18 h) relative to control. STR and SGD transcripts were upregulated after elicitation. H2O2-treated roots showed higher levels of STR at 48-192 h and SGD at 24-48 h, while BSO-JA treatments showed STR increased at 12 h and SGD at 24 h. Also, LC/ESI-MS confirmed the biosynthesis of dolichantoside from N-ω-methyltryptamine and secologanin by U. tomentosa protein extracts.
AB - The activity and gene expression of strictosidine-related enzymes in Uncaria tomentosa root cultures exposed to oxidative stress were studied. Elicitation with 0.2 mM hydrogen peroxide (H2O2) or a combination of 0.8 mM buthionine sulfoximine and 0.2 mM jasmonic acid (BSO-JA) increased peroxidase activities by twofold at Day 8 and glutathione reductase by 1.4-fold at Day 5 in H2O2 elicited cultures respect to the control. Production of monoterpenoid oxindole alkaloids (MOA), 3α-dihydrocadambine, and dolichantoside was stimulated after H2O2 elicitation, reaching levels of 886.4±23.6, 847.7±25.4, and 87.5±7.2 μg/g DW, at Day 8 which were 1.7-, 2.1-, and 2.3-fold higher relative to control. BSO-JA elicited cultures produced about twice alkaloids than H2O2-treated cultures, following a biphasic pattern with maxima at 0.5 and 8 days. Alkaloid production was preceded by increase in strictosidine synthase (STR) and strictosidine glucosidase (SGD) activities. After elicitation with H2O2 or BSO-JA, the STR activity (pKat/mg protein) increased by 1.9-fold (93.8±17.8 at 24 h) or 2.5-fold (102.4±2.2 at 6 h) and the SGD activity (pKat/mg protein) by 2.8-fold (245.2±14.4 at 6 h) or 4.2-fold (421.2±1.8 at 18 h) relative to control. STR and SGD transcripts were upregulated after elicitation. H2O2-treated roots showed higher levels of STR at 48-192 h and SGD at 24-48 h, while BSO-JA treatments showed STR increased at 12 h and SGD at 24 h. Also, LC/ESI-MS confirmed the biosynthesis of dolichantoside from N-ω-methyltryptamine and secologanin by U. tomentosa protein extracts.
KW - Oxidative stress
KW - Oxindole alkaloids
KW - Strictosidine glucosidase
KW - Strictosidine synthase
KW - Uncaria tomentosa
UR - http://www.scopus.com/inward/record.url?scp=84879267886&partnerID=8YFLogxK
U2 - 10.1002/btpr.1723
DO - 10.1002/btpr.1723
M3 - Artículo
C2 - 23606578
SN - 8756-7938
VL - 29
SP - 621
EP - 630
JO - Biotechnology Progress
JF - Biotechnology Progress
IS - 3
ER -