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  • 1
    Electronic Resource
    Electronic Resource
    Isolation of protoplasts and vacuoles from cell suspension cultures of Coleus blumei Benth. (1993)
    Springer
    Plant cell reports 12 (1993), S. 510-512 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to study the accumulation and transport of rosmarinic acid in suspension cells of Coleus blumei we established an efficient method to isolate protoplasts and vacuoles. Protoplasts were disrupted by an osmotic shock in a medium with basic pH containing ethylenediamine tetraacetic acid. The resulting vacuoles were purified on a two-step Ficoll gradient. The comparison of the rosmarinic acid contents of cells, protoplasts and vacuoles showed that the depside is localized in the vacuole. Data concerning the yield and purity of the vacuoles are presented. In addition we show that at the physiological pH of the cytoplasm rosmarinic acid is present almost exclusively as an anion and cannot pass a membrane by simple diffusion. We therefore propose a carrier system for the transport of rosmarinic acid into the vacuole.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00236097
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  • 2
    Electronic Resource
    Electronic Resource
    Purification of rosmarinic acid synthase from cell cultures of Coleus blumei Benth (1993)
    Springer
    Planta 191 (1993), S. 18-22 
    Details
    ISSN: 1432-2048
    Keywords: Coleus (cell cultures) ; Hydroxycinnamic acid ester ; Rosmarinic acid ; Rosmarinic acid synthase (purification)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rosmarinic acid synthase from cell cultures of Coleus blumei Benth. was purified to apparent homogeneity by fractionated ammonium sulfate precipitation (60–80% saturation), hydrophobic interaction chromatography, affinity chromatography and gel filtration. This purification procedure resulted in a 225-fold-enriched specific enzyme activity with a yield of 9%. The protein preparation was apparently pure according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis. The apparent molecular mass determined by gel filtration and SDS-PAGE was 77 kDa, indicating that rosmarinic acid synthase is a monomeric enzyme.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00240891
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  • 3
    Electronic Resource
    Electronic Resource
    The biosynthesis of rosmarinic acid in suspension cultures of Coleus blumei (1994)
    Springer
    Plant cell, tissue and organ culture 38 (1994), S. 171-179 
    Details
    ISSN: 1573-5044
    Keywords: Coleus blumei (suspension cultures) ; hydroxycinnamic acid esters ; phenylalanine ; rosmarinic acid ; secondary metabolism ; tyrosine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Suspension cultures of Coleus blumei accumulate very high amounts of rosmarinic acid, an ester of caffeic acid and 3,4-dihydroxyphenyllactate, in medium with elevated sucrose concentrations. Since the synthesis of this high level of rosmarinic acid occurs in only five days of the culture period, the activities of the enzymes involved in the biosynthesis are very high. Therefore all the enzymes necessary for the formation of rosmarinic acid from the precursors phenylalanine and tyrosine could be isolated from cell cultures of Coleus blumei: phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase, hydroxycinnamoyl:CoA ligase, tyrosine aminotransferase, hydroxyphenylpyruvate reductase, rosmarinic acid synthase and two microsomal 3- and 3′-hydroxylases. The main characteristics of these enzymes of the proposed biosynthetic pathway of rosmarinic acid will be described.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033875
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  • 4
    Electronic Resource
    Electronic Resource
    Influence of the carbon source on growth and rosmarinic acid production in suspension cultures of Coleus blumei (1993)
    Springer
    Plant cell, tissue and organ culture 34 (1993), S. 183-190 
    Details
    ISSN: 1573-5044
    Keywords: carbon source ; cell cultures ; Coleus blumei (Lamiaceae) ; culture medium ; rosmarinic acid ; secondary product formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Suspension cultures of Coleus blumei were characterized with respect to growth and rosmarinic acid formation in media with different sugars and various sugar concentrations. Sucrose is the sugar with the highest stimulating effect on growth and rosmarinic acid accumulation, followed by glucose and fructose. The sugar alcohol mannitol cannot be metabolized by the plant cells. Sucrose is cleaved into glucose and fructose by the Coleus cells. Sucrose concentrations from 1 to 5% have an increasing positive effect on growth and rosmarinic acid synthesis in the cell cultures with a maximum rosmarinic acid content of 12% of the dry weight in medium with 5% sucrose; in medium with 6% sucrose rosmarinic acid accumulation obviously did not reach its highest level in the culture period of 14 days. A very high yield of rosmarinic acid (2 mg ml-1 suspension) could also be achieved by maintaining a sucrose concentration of 2% during the whole culture period. The start of rosmarinic acid synthesis by the cell cultures seems to be regulated by the growth limitation when a nutrient, e.g. phosphate is depleted from the medium. The rate of rosmarinic acid accumulation is related to the amount of carbon left in the medium when growth ceases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00036100
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