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  • 11
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Habituated and tumorous Catharanthus roseus cells grown in the absence of hormones accumulated indole alkaloids. Total alkaloids and alkaloid pattern were the same when cells were cultured in medium without hormones or in alkaloid production medium with and without indole acetic acid. Treatment of cells with Pythium homogenate as elicitor did not increase total alkaloids or change the pattern of alkaloids produced. When either habituated or tumorous cells were grown in 1B5 medium after Gamborg et al (1968) containing 2,4-dichlorophenoxyacetic acid (2,4-D), their capacity to accumulate alkaloids decreased with time. The levels of tryptophan decarboxylase (TDC) and strictosidine synthase (SS) specific activities were constant throughout growth except when cells were exposed to 2,4-D in 1B5 medium, where enzyme activities declined in step with the decrease in alkaloid accumulation. Neither habituated nor tumorous cell suspension cultures accumulated vindoline, nor could they be induced to produce this alkaloid by any of the given treatments.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 42 (1995), S. 81-90 
    ISSN: 1573-5044
    Keywords: Adhesion ; bioreactor ; immobilization ; plant cell culture ; secondary metabolites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Surface immobilization of plant cells avoids the problem of hydrodynamic or shear stress, which tends to be characteristic of suspended cells cultured in typical, mechanically agitated bioreactor systems. Surface immobilization also promotes the natural tendency for plant cells to aggregate, which may improve the synthesis and accumulation of secondary metabolites. In addition, exchange of medium is made simple in surface-immobilized systems, and extracellular secondary products are easily recovered on a continuous basis. However, problems related to regulation of the thickness of the immobilized cell layer, maintenance of the biomass in a productive condition, and vacuolar retention of secondary products have yet to be resolved satisfactorily. This review focusses on two surface-immobilization technologies, differing primarily in the nature and the configuration of the inert support. Prototypes of these designs have been applied to a variety of plant cell systems at bioreactor volumes up to 20 litres. Results obtained with several alternative technologies are also summarized.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 8 (1987), S. 177-182 
    ISSN: 1573-5044
    Keywords: vinca major ; cell suspension ; culture ; chlorophyll content ; indole alkaloids ; strictosidine lactam
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cell lines of Vinca major L. cv. variegata produced strictosidine lactam as the main alkaloid. Quantities varied from 500–1000 μg/g DW in the high-yielding line and 1–100 μg/g DW in the low-yielding line. Transfer of cells to alkaloid production medium resulted in a 6–8 fold increase in alkaloid production with the high-yielding line, some increase in the low-yielding line and in both lines induced intense greening (up to 200 μg chlorophyll/g DW) indicating chloroplast differentiation. Though in the source plant leaves are the main storage site, no correlation between alkaloid accumulation and chloroplast differentiation could be found.
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  • 14
    ISSN: 1573-5044
    Keywords: Catharanthus ; elicitors ; PAL ; TDC ; phenols ; alkaloids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell suspension cultures (cell line No 615) of Catharanthus roseus cv. Little Delicata responded to elicitor treatment by accumulating monoterpenoid indole alkaloids and phenolic compounds. The excretion of phenols into the culture medium resulted from the induction of the branch-point enzyme phenylalanine ammonia lyase. The accumulation of alkaloids, however, occurred several hours earlier than the elicitor-mediated induction of tryptophan decarboxylase through which shikimate pathway intermediates are channelled into tryptamine and related indole alkaloids. The results indicate that both pathways for phenol and indole alkaloid biosynthesis responded to elicitor treatment and that no obvious causal relationship between pathways could be deduced from this study.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 18 (1989), S. 79-93 
    ISSN: 1573-5044
    Keywords: Agrobacterium rhizogenes ; Catharanthus roseus ; hairy root cultures ; indole alkaloids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hairy root cultures of Catharanthus roseus were established by infection with six different Agrobacterium rhizogenes strains. Two plant varieties were used and found to exhibit significantly different responses to infection. Forty-seven hairy root clones derived from normal plants and two derived from the flowerless variety were screened for their growth and indole alkaloid production. The growth rate and morphological appearance showed wide variations between the clones. The alkaloid spectra observed were qualitatively but not quantitatively very similar to that of the corresponding normal plant roots. No vindoline or deacetyltransferase activity could be detected in any of the cultures studied. O-acetylval-lesamine, an alkaloid which has not been previously observed in C. roseus was identified from extracts of hairy root clone No. 8. Two root clones were examined for their growth and alkaloid accumulation during a 26-day culture period. Alkaloid accumulation parallelled growth in both clones with ca. 2 mg ajmalicine and catharanthine per g dry weight being observed.
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  • 16
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 35 (1990), S. 660-667 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A technique was developed to surface immobilize plant cells and was scaled up in laboratory size bioreactors. This technique was shown not to hinder the biosynthetic potential of Catharanthus roseus immobilized cells and to induce a partial release (300 μg/L) of serpentine into the culture medium contrary to suspension cultured cells. The release pattern seemed to follow the biosynthesis trends of the product. This release mechanism could be stimulated by a factor of 10 within 2 h by increasing the pH of the culture from 5.0 to 5.5.
    Additional Material: 5 Ill.
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  • 17
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 33 (1989), S. 293-299 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel technique has been developed to immobilize plant cells. The cells are deposited on a surface of manmade fibrous material that provides for strong binding of the plant tissue biomass growing in the submerged culture. The immobilized plant cells remain fully viable. Relatively uniform biomass loadings of up to 20 mg d.w. plant cells/cm2 support material have been attained. All plant cells from the inoculum suspension became attached within the first 24-48 h depending on the support matrix configuration and hydraulic culture conditions. The advantages and scale-up potential of this technique are discussed and compared to other culturing modes.
    Additional Material: 5 Ill.
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  • 18
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 4 (1962), S. 23-36 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The ability of a number of Ustilago species, especially Ustilago maydis (DC.) Cda., to produce lysine and threonine was investigated. The organisms were grown in shake flasks or in 10-l. fermentors. Lysine and threonine were found to be excreted into the medium both in the free and bound form. The bound amino acids could be released by acid hydrolysis or by enzymes from autolyzed cells. The optimal conditions for the release by autolysis were, in the case of Ustilago maydis (DC.) Cda., pH 4.3 and 45°C. An enzyme that could liberate lysine from the bound form(s) occurring in the broth was extracted from cells of Ustilago maydis (DC.) Cda. It exhibited an apparent pH optimum near 4.0. The effect of pH and temperature during the growth phase on the yield of lysine and threonine was studied in 10-l. fermentations.
    Additional Material: 5 Ill.
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  • 19
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The ability of Ustilago maydis (DC.) Cda. to grow and to produce lysine and threonine was investigated in shake flask cultures. Growth and production of lysine and threonine increased markedly when aeration was increased. The optimal ratio of glucose to diammonium phosphate in the medium seemed to be approximately 10:1. Ammonium phosphate, ammonium sulfate, ammonium chloride, ammonium nitrate, sodium nitrate, urea, aspartic acid, glutamic acid, and glycine were readily used as nitrogen sources. Growth and amino acid production was poor on ammonium carbonate, ammonium acetate, ammonium citrate, trimethylamine, and betaine.U. maydis (DC.) Cda. was found to grow on a number of different carbohydrates. Besides D-glucose the organism could utilize D-mannose, D-galactose, D-fructose, L-arabinose, D-xylose, D-ribose, sucrose, maltose, and the polyalcoholes D-sorbitol, D-mannitol, and i-inositol. Pectin, dextrin, and corn starch treated with α-amylase could also be used but not untreated corn starch or lactose, cellobiose, D-sedoheptulose, glycerol, or D-glucosamine. The formation of lysine and threonine was better with disaccharides and hexoses than with pentoses.No specific effects on the formation of lysine and threonine could be observed from changes in the concentration of calcium, sodium, potassium, magnesium, zinc, iron, copper, manganese, cobalt, or molybdenum. The requirement of metal ions for growth is discussed.
    Additional Material: 2 Ill.
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  • 20
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 35 (1990), S. 702-711 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The scaleup of the technique of plant cell surface immobilization was performed successfully in specifically designed laboratory size bioreactors. The immobilizing matrix was formed into a vertically wound spiral providing for a high immobilizing area-to-volume ratio (0.8-1.2 cm-1). A modified airlift and a mechanically stirred vessel delivered a best bioreactor performance characterized by low biomass frothing and highly efficient plant cell attachment and retention (≥96%). The growth of Catharanthus roseus cells investigated in these bioreactors was found not to be mass transfer limited. It required mild mixing and aeration levels (kLa ∼ 10-15 h-1). The biomass formation pattern of surface immobilized plant cells generally exhibited a linear growth phase followed by a stationary phase characterized by the presence of residual carbohydrates in the medium, contrary to suspension cultures. This behavior was found to depend on the plant cell type and/or line cultured, as well as on the inoculum age. The space restriction and unidirectional growth of the SIPC biofilm combined with the limited availability of essential intracellular nutrients rapidly accumulated from the medium by the stationary phase inoculated plant cells all likely contributed to the culture behavior.
    Additional Material: 6 Ill.
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