Regulatory role of microsomal 3-hydroxy-3-methylglutaryl-coenzyme A reductase for shikonin biosynthesis in Lithospermum erythrorhizon cell suspension cultures

Lange, B. Markus; Severin, Klaus; Bechthold, Andreas; Heide, Lutz

doi:10.1007/s004250050252

Cited by 52 publications

(40 citation statements)

References 46 publications

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“…The expression of C4H2 seemed to be not involved in the light negative regulation of shikonin biosynthesis. The HMGR and LePGT enzymes have been reported to play a significant role in the regulation of shikonin biosynthesis, and white light was shown to strongly suppress HMGR and LePGT1 gene expression in L. erythrorhizon cell cultures (Lange et al, 1998;Yazaki et al, 2002). In contrast to the previous results, the transcripts of HMGR was slightly stimulated under continuous blue and white light.…”

Section: Discussioncontrasting

confidence: 75%

“…At present, the metabolic pathway of shikonin biosynthesis is well-characterized (Heide et al, 1989;Yamaga et al, 1993;Yazaki et al, 1999). Moreover, some genes, PAL (Yazaki et al, 1997b), C4H (Yamamura et al, 2001), 4CL (Yazaki et al, 1995), LePGT , HMGR (Lange et al, 1998), CYP98A6 (Matsuno et al, 2002), and LEPS-2 (Yamamura et al, 2003), encoding metabolic enzymes closely related in shikonin formation and secretion and five genes, LDI1-LDI5 (Yazaki et al, 1997a(Yazaki et al, , 1999(Yazaki et al, , 2001, preferentially expressed in the dark were cloned and identified from the cultured cells. Therefore, the cell cultures of shikonin formation may become one of the promising model systems suitable for biochemical and molecular study of the regulation of plant secondary metabolism.…”

Section: Introductionmentioning

confidence: 98%

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Effect of Light on Gene Expression and Shikonin Formation in Cultured Onosma Paniculatum Cells

Zhi

Chen

et al. 2005

Plant Cell Tiss Organ Cult

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The cell culture in a production medium in the dark is capable of producing a large quantity of shikonin and its derivatives, which were completely inhibited when the cell cultures were irradiated with continuous white light and blue light and partially repressed under continuous red light. The expressions of PAL1, 4CL1, and CYP98A6 were induced under continuous red, blue and white light. Transcript levels of HMGR and LDI2 gradually were decreased in the dark and under continuous red light, while no transcriptional product of LDI2 was detected under blue and white light. C4H2 and LePGT1 genes were constantly expressed irrespective of different light conditions. Among the genes studied, the expression of LDI2 was critical for light-regulated shikonin formation. The inducible expression of PAL1, 4CL1, and CYP98A6 as well as the inhibitory transcription of LDI2 mediated by continuous red light irradiation were likely accounted for the reduced accumulation of shikonin.

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Section: Discussioncontrasting

confidence: 75%

Section: Introductionmentioning

confidence: 98%

Effect of Light on Gene Expression and Shikonin Formation in Cultured Onosma Paniculatum Cells

Zhi

Chen

et al. 2005

Plant Cell Tiss Organ Cult

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“…White or blue light strongly inhibits shikonin production by suppressing the gene expression of PHB geranyltransferase, a key enzyme in shikonin biosynthesis, 41,42) as well as 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, an enzyme responsible for the formation of the geranyl side chain of shikonin. 43) As shown in Fig. 5, LAB production was slightly stimulated by white light irradiation compared with that in cells cultured in the dark, whereas shikonin production was completely inhibited.…”

Section: ϫmentioning

confidence: 72%

Regulation of Lithospermic Acid B and Shikonin Production in Lithospermum erythrorhizon Cell Suspension Cultures.

Yamamoto

Zhao

Yazaki

et al. 2002

CHEMICAL & PHARMACEUTICAL BULLETIN

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Caffeic acid oligomers such as rosmarinic acid and lithospermic acid B (LAB) are distributed in plants of Labiatae and Boraginaceae. Among these oligomers, LAB, the caffeic acid tetramer, has a variety of attractive pharmacological activities such as antioxidant and radical scavenging, 1,2) improvement of renal disease, [3][4][5][6] hypotensive, [7][8][9] improvement of myocardial infarction, 1,10) amplification of beating of myocardial muscles, 11) anti-hepatitis, 12) and aldose reductase inhibitory activity. 13) To utilize LAB as a medicinal resource, more than 30 plant sources were surveyed and only two medicinal plants, Salvia officinalis and Lithospermum erythrorhizon SIEB. et ZUCC, were found to contain this substance in an appreciable amount.14) Many attempts to produce this compound in high quantities using cell or hairy root cultures have been unsuccessful. [15][16][17][18][19][20][21][22] Recently, we found that L. erythrorhizon cell-suspension cultures, which were established for the industrial production of shikonin, 23) produced four caffeic acid oligomers, rosmarinic acid, LAB, a monoglucoside of LAB, and (ϩ)-rabdosiin. 24) In particular, the production of LAB in the cultured cells was highly stimulated in shikonin production medium M-9, 25) reaching almost the same amount as that of shikonin (LAB: ca. 6-10% of dry wt, and total shikonin derivatives: ca. 6-12% of dry wt.). 24,26) Since both LAB and shikonin are formed through a common phenylpropanoid pathway (Chart 1), the counterbalance between two metabolic routes either to LAB or to shikonin would be regulated in L. erythrorhizon cells, suggesting that the selection of suitable regulatory factors would result in higher production of LAB. However, the metabolic linkage and the regulatory mechanism for the biosyntheses of phenylpropanoid-derived compounds of diverse types in L. erythrorhizon cell cultures remain to be clarified. In this study, we investigated the critical regulatory factors to control these two biosynthetic pathways separately and to produce LAB efficiently in the culture system of L. erythrorhizon. ExperimentalCell Cultures The culture strain M18TOM 24) of Lithospermum erythrorhizon, which had been maintained in Linsmaier-Skoog (LS) medium 27) containing IAA 1 mM and kinetin 10 mM, was used in the present study. Cellsuspension cultures in 100-ml flasks each containing 30 ml of medium were agitated on a rotary shaker (80 rpm) at 23°C in the dark and subcultured at intervals of 2 weeks. The cells cultured in LS medium for 2 weeks were inoculated into M-9 medium, 25) modified M-9 or LS media containing 3% sucrose, and the same growth regulators as above (inoculum size: 1 g of fresh cells/30 ml of medium in a 100-ml flask) and cultured for 3 weeks under the above-mentioned culture conditions. For the investigation of the effect of light color on the production of secondary metabolites, each flask was covered with a commercially available colored cellophane sheet, 28) and irradiated with white light (10000 lx) from fluorescent lamps d...

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“…HMGCoA reductase, which regulates the mevalonate pathway in many organisms, was found to catalyze a rate-limiting reaction for GPP biosynthesis in L. erythrorhizon (Köhle et al 2002;Lange et al 1998). The regulatory pattern of gene expression was similar to that of PGT.…”

Section: Biosynthetic Pathway and Its Regulationmentioning

confidence: 73%

<i>Lithospermum erythrorhizon</i> cell cultures: Present and future aspects

Yazaki

2017

Plant Biotechnology

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Lithospermum erythrorhizon cell cultures have been used to produce plant secondary metabolites, as well as in biosynthetic studies. Shikonin, a representative secondary metabolite of L. erythrorhizon, was first produced industrially by dedifferentiated cell cultures in the 1980s. This culture system has since been used in research on various plant secondary metabolites. Other boraginaceaeous plant species, including Arnebia, Echium, Onosma and Alkanna, have been shown to produce shikonin, and studies have assessed shikonin regulation, including transgene expression, in these plants. This review summarizes current knowledge of shikonin production by L. erythrorhizon cell and hairy root cultures, including the historical aspect of large-scale production, and discusses future biochemical and biological research using this species.

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Regulatory role of microsomal 3-hydroxy-3-methylglutaryl-coenzyme A reductase for shikonin biosynthesis in Lithospermum erythrorhizon cell suspension cultures

Cited by 52 publications

References 46 publications

Effect of Light on Gene Expression and Shikonin Formation in Cultured Onosma Paniculatum Cells

Effect of Light on Gene Expression and Shikonin Formation in Cultured Onosma Paniculatum Cells

Regulation of Lithospermic Acid B and Shikonin Production in Lithospermum erythrorhizon Cell Suspension Cultures.

<i>Lithospermum erythrorhizon</i> cell cultures: Present and future aspects

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