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Plant Physiol, November 1999, Vol. 121, pp. 821-828
Cloning and Functional Expression of a Cytochrome P450 cDNA
Encoding 2-Hydroxyisoflavanone Synthase Involved in Biosynthesis of the
Isoflavonoid Skeleton in Licorice1
Tomoyoshi
Akashi,2
Toshio
Aoki, and
Shin-ichi
Ayabe*
Department of Applied Biological Science, Nihon University,
Fujisawa, Kanagawa 252-8510, Japan
Isoflavonoids
are distributed predominantly in leguminous plants and play critical
roles in plant physiology. A cytochrome P450 (P450),
2-hydroxyisoflavanone synthase, is the key enzyme in their
biosynthesis. In cultured licorice (Glycyrrhiza echinata L., Fabaceae) cells, the production of both an isoflavonoid-derived phytoalexin (medicarpin) and a retrochalcone (echinatin) is rapidly induced upon elicitation. In this study, we obtained a full-length P450
cDNA, CYP Ge-8 (CYP93C2), from the cDNA library of elicited G.
echinata cells. When the flavanones liquiritigenin and
naringenin were incubated with the recombinant yeast microsome
expressing CYP93C2, major products emerged and were readily converted
to the isoflavones daidzein and genistein by acid treatment. The chemical structures of the products from liquiritigenin
(2-hydroxyisoflavanone and isoflavone) were confirmed by mass
spectrometry. CYP93C2 was thus shown to encode
2-hydroxyisoflavanone synthase, which catalyzes the
hydroxylation associated with 1,2-aryl migration of flavanones. Northern-blot analysis revealed that transcripts of CYP93C2, in addition to those of other P450s involved in phenylpropanoid/flavonoid pathways, transiently accumulate upon elicitation.
1
This work was supported by a Grant-in-Aid
(no. 09640782) from the Ministry of Education, Sports, Science
and Culture of Japan. T. Akashi was also supported by a research
fellowship (no. 3,883) from the Japan Society for the Promotion of
Science for Young Scientists.
2
Present address: Department of Chemistry,
Faculty of Science, Tokyo Institute of Technology, O-okayama,
Meguro-ku, Tokyo 152-8551, Japan.
*
Corresponding author; e-mail ayabe{at}brs.nihon-u.ac.jp; fax
81-466-80-1141.
© 1999 American Society of Plant Physiologists
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