Journal Article

High Level Expression of Chorismate Pyruvate-Lyase (UbiC) and HMG-CoA Reductase in Hairy Root Cultures of <i>Lithospermum erythrorhizon</i>

Annegret Köhle, Susanne Sommer, Kazufumi Yazaki, Albert Ferrer, Albert Boronat, Shu-Ming Li and Lutz Heide

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 8, pages 894-902
Published in print August 2002 | ISSN: 0032-0781
Published online August 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf106
High Level Expression of Chorismate Pyruvate-Lyase (UbiC) and HMG-CoA Reductase in Hairy Root Cultures of Lithospermum erythrorhizon

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Shikonin, a red naphthoquinone pigment, is produced by cell cultures of Lithospermum erythrorhizon (Boraginaceae). It is biosynthetically derived from two key precursors, 4-hydroxybenzoate (4HB) and geranyldiphosphate (GPP). The bacterial ubiC gene, encoding chorismate pyruvate-lyase (CPL) which converts chorismate to 4-hydroxybenzoate, was expressed in L. erythrorhizon under the control of the strong (ocs)3mas-promoter. This introduced an efficient biosynthetic pathway to 4HB, i.e. a one-step reaction from chorismate, in addition to the endogeneous multi-step phenylpropanoid pathway. Feeding experiments with [1,7-13C2]shikimic acid showed that in the most active transgenic line, 73% of 4HB was synthesized via the genetically introduced pathway. However, there was no correlation between CPL activity and 4HB glucoside or shikonin accumulation in the transgenic lines. HMG-CoA reductase (HMGR) is involved in the biosynthesis of GPP in L. erythrorhizon. Two forms of HMGR1 of Arabidopsis thaliana were expressed in Lithospermum under control of the (ocs)3mas promoter. Only moderate increases in enzyme activity were obtained with the complete enzyme, but high activity was achieved using the soluble cytosolic domain of HMGR1. Shikonin accumulation remained unchanged even upon high expression of soluble HMGR.

Keywords: Keywords: Chorismate pyruvate-lyase — Genetic engineering — HMG-CoA reductase — Lithospermum erythrorhizon — Plant promoter — Shikonin.; Abbreviations: CPL, chorismate pyruvate-lyase; GBA, 3-geranyl-4-hydroxybenzoic acid; GPP, geranyldiphosphate; 4HB, 4-hydroxybenzoic acid; 4HBOG, 4HB glucoside; HMGR, 3-hydroxy-3-methylglutaryl-coenzyme A reductase.

Journal Article.  6284 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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