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  • 1. Barry, S
    et al.
    Brodelius, Peter
    UNIV LUND, CTR CHEM, DEPT PURE & APPL BIOCHEM, S-22007 LUND 7, SWEDEN .
    Mosbach, K
    General Ligand Affinity Chromatography: N6-(6-Aminohexyl) 3',5'-ADP Sepharose as an Affinity Adsorbent for the CoA-Dependent Enzyme, Succinate Thiokinase1976In: FEBS letters, Vol. 70, no 1, p. 261-266Article in journal (Refereed)
  • 2. Bramble, J L
    et al.
    Graves, D J
    Brodelius, Peter
    Department of Plant Biotechnology, University of Lund.
    Calcium and Phosphate Effects on Growth and Alkaloid Production in Coffea arabica: Experimental Results and Mathematical Model.1991In: Biotechnology and Bioengineering, ISSN 0006-3592, E-ISSN 1097-0290, Vol. 37, no 9, p. 859-868Article in journal (Refereed)
    Abstract [en]

    Plant, mammalian, and microbial cells are commonly immobilized in calcium alginate gels for the production of valuable secondary metabolites. However, calcium ions are known to inhibit growth in various type of cells, and calcium is an integral part of such gels. Therefore, an investigation was conducted to evaluate the effect of calcium on the growth and alkaloid production of a model cell-line, Coffea arabica, in suspension culture before attempting to immobilize such cells in alginate. A kinetic model was then developed from the results to describe cell growth and alkaloid production and the mechanism by which calcium influences these variables. In addition, it was observed that there was a characteristic relationship between the concentration of calcium in the external medium and the concentration of extracellular and intracellular phosphate. The intracellular phosphate level was, in turn, related to the production of alkaloids. Using these results, a dynamic mathematical model of cell growth and alkaloid production was developed based on the proposed roles of calcium and phosphate. The model showed satisfactory agreement with three sets of experiments at different calcium concentrations. A possible linkage between the calcium and phosphate results is postulated based on the limited solubility of calcium phosphate. 

  • 3. Bramble, J L
    et al.
    Graves, D J
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Plant Cell Culture using a Novel Bioreactor: The Magnetically Stabilized Fluidized Bed1990In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 6, no 6, p. 452-457Article in journal (Refereed)
    Abstract [en]

    A novel bioreactor using magnetically stabilized fluidized bed (MSFB) technology has been developed that has certain advantages for cultivating cells continuously. In this system, the cells are protected from shear and are constrained to move through the fermenter in lock-step fashion by being immobilized in calcium alginate beads. The MSFB permits good mass transfer, minimizes particle collisions, and allows for the production of cells while maintaining a controlled cell residence time. Details of the experimental system are described. In addition, the experimental performance of an MSFB used to grow plant cells in batch mode is compared to the results obtained in shake flask culture. 

  • 4.
    Brodelius, Maria
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Hiraiwa, M
    Martilla, S
    Picaud, S
    Al-Karadaghi, S
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Immunolocalization of the saposin-like insert of plant aspartic proteinases exhibiting saposin C activity in seeds and young flower tissues2005In: Physiologia Plantarum, Vol. 125, p. 405-418Article in journal (Refereed)
  • 5.
    Brodelius, Maria
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Lundgren, Anneli
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Mercke, Per
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Fusion of farnesyldiphosphate synthase and epi-aristolochene synthase, a sesquiterpene cyclase involved in capsidiol biosynthesis in Nicotiana tabacum2002In: European Journal of Biochemistry, ISSN 0014-2956, E-ISSN 1432-1033, Vol. 269, p. 3570-3577Article in journal (Refereed)
  • 6.
    Brodelius, Peter
    Department of Plant Biochemistry, University of Lund .
    Enzyme assays1991In: Current Opinion in Biotechnology, ISSN 0958-1669, E-ISSN 1879-0429, Vol. 2, no 1, p. 23-29Article in journal (Refereed)
    Abstract [en]

    The past year or so has seen the development of new enzyme assays, as well as the improvement of existing ones. Assays are becoming more rapid and sensitive as a result of modifications such as amplification of the enzyme product(s). Recombinant DNA technology is now being recognized as a particularly useful tool in the search for improved assay systems. 

  • 7.
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    High-performance Liquid Chromatographic Analysis of Analogous Amino and Oxo Acids for the Determination of Amino Acid Oxidase and Transaminase Activities1984In: Acta chemica Scandinavica, Series B - Organic Chemistry and Biochemistry, Vol. 38, no 3, p. 219-223Article in journal (Refereed)
  • 8.
    Brodelius, Peter
    Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Ziirich, Switzerland.
    Permeabilization of Plant Cells for Release of Intracellularly Stored Products: Viability Studies1988In: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 27, p. 561-566Article in journal (Refereed)
    Abstract [en]

    The effects of various chemical substanceson the permeability of plasma membranesand tonoplasts of three suspension cultures (Catharanthusroseus, Thalictrum rugosum and Chenopodiumrubrum) have been studied. The permeabilityof the plasma membrane is monitoredby measuring the activity of the cytosolic enzymeisocitrate dehydrogenase and the permeability ofthe tonoplast is measured by determining the releaseof substances stored in the vacuoles (inorganicphosphate, berberine and betanin for thethree cell lines, respectively). The minimum concentrationrequired for quantitative release of vacuolarproducts have been established for five differentpermeabilization agents. Cell viability islost upon permeabilization except for treatmentof Catharanthus roseus with DMSO and Triton X-100. 

  • 9.
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Phenylpropanoid Metabolism in Vanilla planifolia Andr. V. Formation of Flavour Compounds in Developing Fruits1994In: Phytochemical analysis, Vol. 5, p. 27-31Article in journal (Refereed)
  • 10.
    Brodelius, Peter
    Institute of Biotechnology, Swiss Federal Institute of Technology, Hönggerberg, CH-8093 Zürich, Switzerland.
    The Potential Role of Immobilization in Plant Cell Biotechnology1985In: Trends in Biotechnology, ISSN 0167-7799, E-ISSN 1879-3096, Vol. 3, no 11, p. 280-285Article in journal (Refereed)
    Abstract [en]

    It has long been hoped that the unique biosynthetic capacity of plants could be exploited in vitro using culture systems analogous to microbial fermentations. However, the characteristics of both the growth and metabolism of plant cells in vitro differ considerably from those of microbial cells and plant cell suspension culture systems have met with limited success. Immobilizing the cells creates a new set of options for the plant biotechnologist to explore. Improvements in some process criteria are apparent although evaluating the potential of immobilized plant cells for producing commercial compounds will only be possible when the biological problems have been overcome. 

  • 11.
    Brodelius, Peter
    et al.
    Biochemistry 2, University of Lund, Chemical Center, P. O. B. 740, S-22007 Lund 7, Sweden.
    Deus, B
    Mosbach, K
    Zenk, M H
    Immobilized Plant Cells for the Production and Transformation of Natural Products1979In: FEBS letters, Vol. 103, no 1, p. 93-97Article in journal (Refereed)
  • 12.
    Brodelius, Peter
    et al.
    Institute of Biotechnology ETH-Hönggerberg CH-8093 Zürich, Switzerland.
    Funk, C
    Häner, A
    Villegas, Mirza
    A Procedure for the Determination of Optimal Chitosan Concentrations for Elicitation of Cultured Plant Cells.1989In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 28, no 10, p. 2651-2654Article in journal (Refereed)
    Abstract [en]

    An experimental method for determination of the optimum chitosan concentration for elicitation of plant cell suspension cultures is presented. The procedure, which is based on measurements of the conductivity of the culture medium after addition of chitosan, has been applied to suspension cultures of Nicotiana tabacum and Eschscholtzia californica. Increased conductivity of the medium (due to permeabilization of the cells) results in decreased secondary product formation and cell growth. Maximum product formation is observed for cells elicited with the highest chitosan concentration which does not affect membrane permeability. 

  • 13.
    Brodelius, Peter
    et al.
    Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Ziirich, Switzerland.
    Funk, C
    Shillito, R D
    Permeabilization of Cultivated Plant Cells by Electroporation for Release of Intracellularly Stored Secondary Products1988In: Plant Cell Reports, ISSN 0721-7714, E-ISSN 1432-203X, Vol. 7, no 3, p. 186-188Article in journal (Refereed)
    Abstract [en]

    Plant cell suspension cultures producing secondary metabolites have been permeabilized for product release by electroporation. The two cell cultures studied, i.e. Thalictrum rugosum and Chenopodium rubrum, require about 5 and 10 kV cm–1, respectively, for complete permeabilization (release of all the intracellularly stored product). The number of electrical pulses and capacitance used had a relatively limited effect on product release while the viability of the cells was strongly influenced by the latter. Conditions for complete product release resulted in total loss of viability of the cells after treatment. The release of product from immobilized cells was also achieved by electroporation. Cells entrapped in alginate required less voltage for permeabilization than free or agarose entrapped cells. 

  • 14.
    Brodelius, Peter
    et al.
    Department of Chemistry, Q-058, University of California at San Diego, La Jolla, California 92093, U.S.A.
    Kaplan, N O
    Studies of Bovine Liver Glutamate Dehydrogenase by Analytical Affinity Chromatography on Immobilized AMP Analogs1979In: Archives of Biochemistry and Biophysics, Vol. 194, no 2, p. 449-456Article in journal (Refereed)
    Abstract [en]

    Bovine liver glutamate dehydrogenase has been studied by analytical affinity chromatography on two immobilized AMP analogs, i.e., N6-(6-aminohexyl)-AMP and 8-(6-aminohexyl)-amino-AMP. The existence of various enzyme-coenzyme and enzyme-effector complexes has been verified. Also the cooperative formation of two ternary complexes, i.e., glutamic dehydrogenase (GHD)-NADP-glutamate and GDH-ADP-leucine, has been shown. The results of this study have been rationalized by the “ligand exclusion theory.” which has been proposed for the regulation of the glutamic dehydrogenase. It has been shown that the active site and the ADP-binding effector site are oriented close to each other on the enzyme. Furthermore, the data suggest that the adenylic site is not identical to the nonactive coenzyme binding site. A mechanism based on electrostatic interactions is suggested for the cooperative binding of oxidized coenzyme and substrate. Dissociation constants for complexes between the enzyme and two coenzyme fragments (P-ADPR and 2′,5′-ADP) have been estimated.

  • 15.
    Brodelius, Peter
    et al.
    UNIV LUND, CTR CHEM, DEPT PURE & APPL BIOCHEM, S-22007 LUND 7, SWEDEN .
    Lannom, R A
    Kaplan, N O
    The Synthesis of 8-(6-Aminohexyl)-Amino-GMP and Its Applications as a General Ligand in Affinity Chromatography1978In: Archives of Biochemistry and Biophysics, Vol. 188, no 1, p. 228-231Article in journal (Refereed)
    Abstract [en]

    The synthesis of a new general ligand, i.e., 8-(6-aminohexyl)-amino-GMP, has been achieved by bromination of GMP and subsequent substitution of the bromine for hexamethylene diamine. The overall yield of the synthesis has been 60 to 70%. This new general ligand was immobilized on BrN-activated Sepharose and used as an affinity adsorbent for inosinic acid dehydrogenase (E.C. 1.2.1.14) from Aerobacter aerogenes. Various elution methods were investigated in order to increase the specific activity of the purified enzyme. 

  • 16.
    Brodelius, Peter
    et al.
    Avdelningen for Biokemi, Kemicentrum, Lunds Universitet.
    Larsson, P-O
    Avdelningen for Biokemi, Kemicentrum, Lunds Universitet.
    Mosbach, Klaus
    Avdelningen for Biokemi, Kemicentrum, Lunds Universitet.
    The Synthesis of Three AMP-Analogues: N6-(6-Amino-hexyl)-Adenosine 5'-Monophosphate, N6-(6-Aminohexyl)-Adenosine 2',5'-Bisphosphate, and N6-(6-Aminohexyl)-Adenosine 3',5'-Bisphosphate and Their Application as General Ligands in Biospecific Affinity Chromatography1974In: European Journal of Biochemistry, ISSN 0014-2956, E-ISSN 1432-1033, Vol. 47, p. 81-89Article in journal (Refereed)
    Abstract [en]

    Phosphorylation of 6-chloropurine riboside with phosphorus oxychloride and phosphorus trichloride gave a mixture of the two isomers, 6-chloropurine-riboside 2’,5‘-bisphosphate and 6-chloropurine-riboside 3‘,5‘-bisphosphate. Reaction with Iy6-diaminohexane followed by resolution of the isomeric mixture on a Dowex 1-X2 column yielded N6-(6-aminohexyl)-adenosine 2’,5’-bisphosphate and N6-(6-aminohexyl)-adenosine 3’,5‘-bisphosphate.The inhibition of several NADP+-dependent and NAD+-dependent dehydrogenases by N6-(6-aminohexyl)-adenosine 2’,5‘-bisphosphate, N6-(6-aminohexyl)-adenosine 3’,5’-bisphosphate and N6-(6-aminohexyl)-adenosine 5’-monophosphate was examined.These three AMP-analogues were attached to Sepharose 4B by the cyanogen bromide method and the binding of several NAD(P)+-dependent enzymes were investigated. NADP+-dependent enzymes were bound to Sepharose substituted with N6-(6-aminohexyl)-adenosine 2’,5‘-bisphosphate, whereas NAD+-dependent enzymes were not bound under the same conditions. Conversely, when N6-(6-aminohexy1)-adenosine 5‘-monophosphate was used as the immobilised ligand only the NAD+- dependent enzymes were bound, as well as glucose-6-phosphate dehydrogenase showing weak affinity. These observations strongly suggest that these two immobilised analogues represent true biospecific and group-specific adsorbents. The enzymes were eluted with their complementary nucleotides, NAD(H) and NADP(H). These techniques were utilised to purify several NADPf-dependent enzymes from a crude Candida utilis extract by chromatography on the new biospecific adsorbent.

  • 17.
    Brodelius, Peter
    et al.
    UNIV LUND, CTR CHEM, DEPT PURE & APPL BIOCHEM, S-22007 LUND 7, SWEDEN .
    Mosbach, K
    Determination of Dissociation Constants for Binary Dehydrogenase-Coenzyme Complexes by (Bio)Affinity Chromatography on an Immobilized AMP-Analogue1976In: Analytical biochemistry, Vol. 72, no 1-2, p. 629-636Article in journal (Refereed)
    Abstract [en]

    Various alcohol and lactate dehydrogenases were adsorbed to an affinity column of immobilized N6-(6-aminohexyl)-AMP and subsequently eluted with gradients of coenzymes or coenzyme fragments. A linear relation was observed between the eluting concentration of nucleotide and the reported dissociation constants for the corresponding binary enzyme-nucleotide complexes. This relation has been utilized to determine unknown dissociation constants by affinity chromatography. The method presented can also be utilized for the estimation of dissociation constants betweendehydrogenases and coenzyme analogues. 

  • 18.
    Brodelius, Peter
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Mosbach, K
    The Utilization of Immobilized Substrate/Product in Affinity Chromatography. A Model Study using alfa-Chymotrypsin1973In: Acta chemica Scandinavica, Vol. 27, p. 2634-2638Article in journal (Refereed)
  • 19.
    Brodelius, Peter
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Mosbach, Klaus
    Separation of the Isoenzymes of Lactate Dehydrogenase by Affinity Chromatography Using an Immobilized AMP-Analogue1973In: FEBS Letters, ISSN 0014-5793, E-ISSN 1873-3468, Vol. 35, no 2, p. 223-226Article in journal (Refereed)
  • 20.
    Brodelius, Peter
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Nilsson, K
    Entrapment of Plant Cells in Different Matrices. A Comparative Study1980In: FEBS letters, Vol. 122, no 2, p. 312-316Article in journal (Refereed)
  • 21.
    Brodelius, Peter
    et al.
    Lunds Universitet.
    Nilsson, K
    Permeabilization of Immobilized Plant Cells, Resulting in Release of Intracellularly Stored Products with Preserved Cell Viability1983In: European Journal of Applied Microbiology and Biotechnology, ISSN 0340-2118, Vol. 17, no 5, p. 275-280Article in journal (Refereed)
  • 22.
    Brodelius, Peter
    et al.
    Lunds Universitet.
    Nilsson, K
    Mosbach, K
    Production of alfa-Keto Acids. Part I - Immobilized Cells of Trigonopsis variabilis containing D-Amino Acid Oxidase1981In: Applied biochemistry and biotechnology, Vol. 6, p. 293-308Article in journal (Refereed)
  • 23.
    Brodelius, Peter
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Pedersen, H
    Increasing Secondary Metabolite Production in Plant Cell Cultures by Redirecting Transport1993In: Trends in biotechnology, Vol. 11, p. 30-36Article in journal (Refereed)
  • 24.
    Brodelius, Peter
    et al.
    Institute of Biotechnology, Swiss Federal Institute of Technology, Honggerberg, CH-8093 Zurich, Switzerland.
    Vogel, H J
    A Phosphorus-31 Nuclear Magnetic Resonance Study of Phosphate Uptake and Storage in Cultured Catharanthus roseus and Daucus carota Plant Cells1985In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 260, p. 3556-3560Article in journal (Refereed)
    Abstract [en]

    High resolution "P NMR spectra (103.2 MHz) ofoxygenated Catharanthus roseu8 and Daucus carotacells grown in suspension cultures were obtained usinga solenoidal perfusion probe. The spectra showed resonancesfor various phosphorylated metabolites suchas ATP, ADP, NAD(P)(H), nucleoside diphosphoglucose,and sugar phosphates. The relative levels ofthe phosphorylated metabolites remained constantthroughout the growth curve. No resonances for storagecompounds such as polyphosphates, pyrophosphate,or phytates were observed. Two resolved resonancesfor Pi indicated an intracellular pH of 7.3 and5.7 (or below) for the cytoplasm and vacuoles, respectively.The time course of Pi uptake and storage duringgrowth in fresh culture medium was followed by studyingthe level of vacuolar Pi with 31PN MR (145.7 MHz).Simultaneously, the level of Pi in the culture mediumwas followed with radioactive s2P. C. roseus quicklytakes up all the Pi from the culture medium (maximumrate 1.7 pmol min" g" (dry weight of cells)). The Pi isfirst stored in the vacuoles; subsequently, one part ofthis pool is used to keep a constant cytoplasmic Pi levelwhile another part is apparently accumulated as anNMR invisible Pi store, probably in another cell organelle.In contrast, D. carota does not accumulate Pi inthe vacuoles and consequently it takes up Pi from themedium at a much slower rate (0.05 pmol min" g"(dry weight of cells)). 

  • 25.
    Brodelius, Peter
    et al.
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Xue, Zhong-tian
    Isolation and Characterization of a cDNA from Cell Suspension Cultures of Vanilla planifolia Andr. Encoding 4-Coumarate:CoA Ligase1997In: Plant physiology and biochemistry, Vol. 35, p. 497-506Article in journal (Refereed)
  • 26. C White, Paul
    et al.
    Brodelius, Maria
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Arnold, Daniele
    Kay, John
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Processing, activity, and inhibition of recombinant cyprosin, an aspartic proteinase from Cardoon (Cynara cardunculus)1999In: Journal of biological chemistry, Vol. 274, p. 16685-16693Article in journal (Refereed)
  • 27. Collinge, M A
    et al.
    Brodelius, Peter
    Institute of Biotechnology ETH-Hönggerberg CH-8093 Zürich, Switzerland.
    Dynamics of Benzophenanthridine Alkaloid Production in Suspension Cultures of Eschscholtzia californica after Treatment with a Yeast Elicitor1989In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 28, no 4, p. 1101-1104Article in journal (Refereed)
    Abstract [en]

    A number of benzophenanthridine alkaloids are induced in suspension cultures of Eschscholtzia californica after treatment with an elicitor prepared from yeast extract. The formation of the alkaloids sanguinarine, chelerythrine and macarpine has been studied in relation to; elicitor concentration, incubation time after elicitation, and culture age. A significant portion of these alkaloids is released into the medium. Sanguinarine and chelerythrine reach maximum levels a few hours after the time of elicitation. Thereafter, their levels decline and the amount of macarpine increases. Viability of elicited cells, as determined by their subsequent growth, is not significantly reduced. There is a good correlation between induced tyrosine decarboxylase activity and alkaloid formation. 

  • 28. Cordeiro, M C
    et al.
    Jacob, E
    Puhan, Z
    S Pais, Maria
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Milk Clotting and Proteolytic Activities of Purified Cynarases from Cynara cardunculus L.; A Comparison to Chymosin1992In: Milchwissenschaft, Vol. 47, p. 683-687Article in journal (Refereed)
  • 29. Cordeiro, M C
    et al.
    S Pais, Maria
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Tissue-Specific Expression of Multiple Forms of Cyprosin (Aspartic Proteinase) in Flowers of Cynara cardunculus1994In: Physiologia plantarum, Vol. 92, p. 645-653Article in journal (Refereed)
  • 30. Cordeiro, M C
    et al.
    Xue, Zhong-tian
    Pietrzak, M
    S Pais, Maria
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Isolation and Characterization of a cDNA from Flowers of Cynara cardunculus Encoding Cyprosin (an Aspartic Proteinase) and Its Use to Study the Organ-Specific Expression of Cyprosin1994In: Plant molecular biology, Vol. 24, p. 733-741Article in journal (Refereed)
  • 31. Cordeiro, M C
    et al.
    Xue, Zhong-tian
    S Pais, Maria
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Proteases from Cell Suspension Cultures of Cynara cardunculus1993In: Phytochemistry, Vol. 33, p. 1323-1326Article in journal (Refereed)
  • 32. Domingos, Ana
    et al.
    C Cardoso, Paula
    Xue, Zhong-tian
    Clemente, Alda
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    S Pais, Maria
    Purification, cloning and autoproteolytic processing of an aspartic proteinase from Centaurea calcitrapa2000In: European Journal of Biochemistry, Vol. 267, p. 6824-6831Article in journal (Refereed)
  • 33. Drakenberg, T
    et al.
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    McIntyre, D D
    Vogel, H J
    Structural Studies of Digitoxin and Related Cardenolides by two-dimentional NMR1990In: Canadian journal of biochemistry, ISSN 0008-4018, Vol. 68, p. 272-277Article in journal (Refereed)
  • 34. Felix, H
    et al.
    Brodelius, Peter
    Lunds Universitet.
    Mosbach, K
    Enzyme Activities of the Primary and Secondary Metabolism of Simultaneously Permeabilized and Immobilized Plant Cells1981In: Analytical biochemistry, Vol. 116, no 2, p. 462-470Article in journal (Refereed)
  • 35. Funk, C
    et al.
    Brodelius, Peter
    Institute of Biotechnology ETH-Hönggerberg CH-8093 Zürich, Switzerland.
    Influence of Growth Regulators and an Elicitor on Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia1990In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 29, no 3, p. 845-848Article in journal (Refereed)
    Abstract [en]

    A cell suspension culture of Vanilla planifolia has been established in MS-medium. 2,4-D suppressed while NAA enhanced the formation of extractable phenolics. Cytokinins appeared to favour lignin biosynthesis. Treatment of the culture with chitosan resulted in the induction of various enzymes of phenylpropanoid metabolism, while the amount of extractable phenolics decreased due to their rapid incorporation into polymeric ligneous material. 

  • 36. Funk, C
    et al.
    Brodelius, Peter
    Department of Plant Biochemistry, University of Lund.
    Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia. Andr. II. Effects of Precursor Feeding and Metabolic Inhibitors1990In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 94, p. 95-101Article in journal (Refereed)
    Abstract [en]

    Feeding of cinnamic acid and ferulic acid to non-treated andchitosan-treated cell suspension cultures of Vanilla planifoIiaresulted in the formation of trace amounts of p-hydroxy benzoicacid (5.2 micrograms per gram fresh weight of cells) and vanillicacid (6.4 micrograms per gram fresh weight of cells), respectively.Addition of a 4-hydroxycinnamate: CoA-ligase inhibitor, 3,4-(methylenedioxy)-cinnamic acid (MDCA), resulted in a reducedbiosynthesis of ligneous material with a simultaneous significantincreased vanillic acid formation (around 75 micrograms per gramfresh weight of cells). A K, of 100 micromolar for 4-hydroxycinnamate:CoA-ligase in a crude preparation was estimated for thisinhibitor. It is suggested that the conversion of cinnamic acidsinto benzoic acids does not involve cinnamoyl CoA esters asintermediates. Feeding of 14C-cinnamic acid and 14C-ferulic acidto cells treated with MDCA indicate that cinnamic acid, but notferulic acid, is a precursor of vanillic acid in these cultivated cellsof V. planifolia. 

  • 37. Funk, C
    et al.
    Brodelius, Peter
    Department of Plant Biochemistry, University of Lund.
    Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia. Andr. III. Conversion of 4-Methoxycinnamic acids into 4-Hydroxybenzoic acids1990In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 94, p. 102-108Article in journal (Refereed)
    Abstract [en]

    Feeding of 4-methoxycinnamic acid, 3,4-dimethoxycinnamicacid and 3,4,5-trimethoxycinnamic acid to cell suspension culturesof Vanilla planifolia resulted in the formation of 4-hydroxybenzoicacid, vanillic acid, and syringic acid, respectively. Thehomologous 4-methoxybenzoic acids were demethylated to thesame products. It is concluded that the side chain degradingenzyme system accepts the 4-methoxylated substrates while thedemethylation occurs at the benzoic acid level. The demethylatingenzyme is specific for the 4-position. Feeding of [10_4Cmethyl]-3,4-dimethoxycinnamic acid revealed that the first stepin the conversion is the glycosylation of the cinnamic acid to itsglucose ester. A partial purification of a UDP-glucose: transcinnamicacid glucosyltransferase is reported. 4-Methoxy substitutedcinnamic acids are better substrates for this enzyme than4-hydroxy substituted cinnamic acid. It is suggested that 4-methoxysubstituted cinnamic acids are intermediates in the biosyntheticconversion of cinnamic acids to benzoic acids in cells ofV. planifolia. 

  • 38. Funk, C
    et al.
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia. Andr. IV. Induction of Vanillic Acid Formation1992In: Plant physiology, Vol. 99, p. 256-262Article in journal (Refereed)
  • 39. Funk, C
    et al.
    Gügler, K
    Brodelius, Peter
    Institute of Biotechnology, ETH/Hönggerberg, CH-8093 Zürich, Switzerland.
    Increased Secondary Product Formation in Plant Cell Suspension Cultures after Treatment with a Yeast Carbohydrate (Elicitor)1987In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 26, p. 401-405Article in journal (Refereed)
    Abstract [en]

    A carbohydrate fraction isolated from yeast extract by ethanolic precipitation was used as an elicitor to induce secondary product formation in plant cell suspension cultures. The elicitor preparation is effective in inducing glyceollin isomer synthesis (up to 200 μg glyceollin per g dry wt) in cells of Glycine max and enhancing berberine biosynthesis (up to four-fold) in cells of Thalictrum rugosum. The response of the cell cultures to the elicitor treatment is dependent on the amount of carbohydrate per unit of biomass and on the physiological state of the cells. Cells are optimally induced in late exponential or early stationary growth phases. 

  • 40.
    Gliszczynska, Anna
    et al.
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Brodelius, Peter E.
    Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
    Sesquiterpene coumarins2012In: Phytochemistry Reviews, ISSN 1568-7767, E-ISSN 1572-980X, Vol. 11, no 1, p. 77-96Article in journal (Refereed)
    Abstract [en]

    Plants have a long history as therapeutic tools in the treatment of human diseases and have been used as a source of medicines for ages. In search of new biologically active natural products, many plants and herbs used in traditional medicine are screened for natural products with pharmacological activity. In this paper, we present a group of natural products, the sesquiterpene coumarins isolated from plants, and describe their wide range of biological activity. Sesquiterpene coumarins are found in some plants of the families Apiaceae (Umbelliferae), Asteraceae (Compositae) and Rutaceae. The coumarin moiety is often umbelliferone (7-hydroxycoumarin) but scopo- letin (7-hydroxy-6-methoxycoumarin) and isofraxidin (7-hydroxy-6,8-dimethoxycoumarin) are also found. These coumarins are linked to a C15 terpene moiety through an ether linkage. Another group of sesquiter- pene coumarins is the prenylated 4-hydroxycoumarins where the link between the coumarin and the C15 terpene moiety is a C–C-bond at carbon 3 of the coumarin moiety. Finally, the prenylfurocoumarin-type sesquiterpenoids are a separate group of sesquiterpene coumarins based on the suggested biosynthetic pathway. Our relatively limited knowledge on the biosynthesis of sesquiterpene coumarins is reviewed.

  • 41.
    Guo, Ming
    et al.
    Zhejiang A & F University, China.
    Lu, Xiaowang
    Zhejiang A & F University, China.
    Wang, Yan
    Zhejiang A & F University, China.
    Brodelius, Peter E.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Comparison of the interaction between lactoferrin and isomeric drugs2017In: Spectrochimica Acta Part A - Molecular and Biomolecular Spectroscopy, ISSN 1386-1425, E-ISSN 1873-3557, Vol. 173, p. 593-607Article in journal (Refereed)
    Abstract [en]

    The binding properties of pentacyclic triterpenoid isomeric drugs, i.e. ursolic acid (UA) and oleanolic acid (OA), to bovine lactoferrin (BLF) have been studied by molecule modeling, fluorescence spectroscopy, UV-visible absorbance spectroscopy and infrared spectroscopy (IR). Molecular docking, performed to reveal the possible binding mode or mechanism, suggested that hydrophobic interaction and hydrogen bonding play important roles to stabilize the complex. The results of spectroscopic measurements showed that the two isomeric drugs both strongly quenched the intrinsic fluorescence of BLF through a static quenching procedure although some differences between UM and OA binding strength and non-radiation energy transfer occurred within the molecules. The number of binding sites was 3.44 and 3.10 for UA and OA, respectively, and the efficiency of Forster energy transfer provided a distance of 0.77 and 1.21 nm for UA and OA, respectively. The conformation transformation of BLF affected by the drugs conformed to the "all-or-none" pattern. In addition, the changes of the ratios of alpha-helices, beta-sheets and beta-turns of BLF during the process of the interaction were obtained. The results of the experiments in combination with the calculations showed that there are two modes of pentacyclic triterpenoid binding to BLF instead of one binding mode only governed by the principle of the lowest bonding energy.

  • 42.
    Guo, Ming
    et al.
    Zhejiang Agr & Forestry Univ, Peoples R China.
    Wang, Xiaomeng
    Zhejiang Agr & Forestry Univ, Peoples R China.
    Lu, Xiaowang
    Zhejiang Agr & Forestry Univ, Peoples R China.
    Wang, Hongzheng
    Zhejiang Agr & Forestry Univ, Peoples R China.
    Brodelius, Peter E.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    alpha-Mangostin Extraction from the Native Mangosteen (Garcinia mangostana L.) and the Binding Mechanisms of alpha-Mangostin to HSA or TRF2016In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 11, no 9, article id e0161566Article in journal (Refereed)
    Abstract [en]

    In order to obtain the biological active compound, alpha-mangostin, from the traditional native mangosteen (Garcinia mangostana L.), an extraction method for industrial application was explored. A high yield of a-mangostin (5.2%) was obtained by extraction from dried mangosteen pericarps with subsequent purification on macroporous resin HPD-400. The chemical structure of alpha-mangostin was verified mass spectrometry (MS), nuclear magnetic resonance (H-1 NMR and C-13 NMR), infrared spectroscopy (IR) and UV-Vis spectroscopy. The purity of the obtained alpha-mangostin was 95.6% as determined by HPLC analysis. The binding of native alpha-mangostin to human serum albumin (HSA) or transferrin (TRF) was explored by combining spectral experiments with molecular modeling. The results showed that amangostin binds to HSA or TRF as static complexes but the binding affinities were different in different systems. The binding constants and thermodynamic parameters were measured by fluorescence spectroscopy and absorbance spectra. The association constant of HSA or TRF binding to alpha-mangostin is 6.4832x10(5) L/mol and 1.4652x10(5) L/mol at 298 K and 7.8619x10(5) L/mol and 1.1582x10(5) L/mol at 310 K, respectively. The binding distance, the energy transfer efficiency between alpha-mangostin and HSA or TRF were also obtained by virtue of the Forster theory of non-radiation energy transfer. The effect of alpha-mangostin on the HSA or TRF conformation was analyzed by synchronous spectrometry and fluorescence polarization studies. Molecular docking results reveal that the main interaction between amangostin and HSA is hydrophobic interactions, while the main interaction between alpha-mangostin and TRF is hydrogen bonding and Van der Waals forces. These results are consistent with spectral results.

  • 43. Gügler, K
    et al.
    Dèfago, G
    Brodelius, Peter
    University of Kalmar, School of Pure and Applied Natural Sciences.
    Pathogenesis-related Protein b1" in Plants and in Cell Suspension Cultures of Nicotiana glutinosa, Nicotiana debneyi and an Amphidiploid Cross (N. glutinosa x N. debneyi)1992In: Physiologia plantarum, Vol. 85, p. 1-8Article in journal (Refereed)
  • 44. Gügler, K
    et al.
    Funk, C
    Brodelius, Peter
    Institute of Biotechnology, Swiss Federal Institute of Technology, Hönggerberg, CH-8093 Zfirich, Switzerland.
    Elicitor-induced Tyrosine Decarboxylase in Berberine Synthesizing Suspension Cultures of Thalictrum rugosum1988In: European Journal of Biochemistry, ISSN 0014-2956, E-ISSN 1432-1033, Vol. 170, no 3, p. 661-666Article in journal (Refereed)
    Abstract [en]

    Tyrosine decarboxylase (EC 4.1.1.25) was induced in suspension cultures of Thalictrum rugosum by treatment with a yeast glucan elicitor. Maximum induction was observed at a carbohydrate concentration of 0.4 mg/g fresh weight of cells and maximum enzyme activity was reached 20 h after addition of elicitor. The enzyme was inducible in late exponential and early stationary growth phases. A good correlation between induced tyrosine decarboxylase activity and berberine biosynthesis has been established. It is suggested that tyrosine decarboxylase may be a key enzyme between primary and secondary metabolisms in the biosynthesis of norlaudanosoline-derived alkaloids. 

  • 45. Haldimann, D
    et al.
    Brodelius, Peter
    Institute of Biotechnology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland.
    Redirecting Cellular Metabolism by Immobilization of Cultured Plant Cells: A Model Study with Coffea arabica1987In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 26, no 5, p. 1431-1434Article in journal (Refereed)
    Abstract [en]

    Suspension-cultured cells of Coffea arabica have been immobilized by entrapment in calcium alginate gels to mimic natural aggregation. The production of methylxanthine alkaloid was increased up to 13-fold by the immobilization. This increased production has been ascribed to organization of the entrapped cells through physicochemical interactions between the polymer (alginate) and the plant cell wall. It has been shown that the metabolic changes induced by the immobilization are reversible. 

  • 46.
    Han, Junli
    et al.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Wang, Hongzhen
    Zhejiang Agr & Forestry Univ, Peoples Republic of China.
    Kanagarajan, Selvaraju
    Swedish University of Agricultural Sciences.
    Hao, Mengshu
    Lund University.
    Lundgren, Anneli
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Brodelius, Peter E.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Promoting Artemisinin Biosynthesis in Artemisia annua Plants by Substrate Channeling2016In: Molecular Plant, ISSN 1674-2052, E-ISSN 1752-9867, Vol. 9, no 6, p. 946-948Article in journal (Refereed)
  • 47.
    Han, Junli
    et al.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Wang, Hongzhen
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. Zhejiang Agriculture and Forestry University, Linan 311300, Zhejiang, PR China..
    Lundgren, Anneli
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Brodelius, Peter E.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Effects of overexpression of AaWRKY1 on artemisinin biosynthesis in transgenic Artemisia annua plants2014In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 102, p. 89-96Article in journal (Refereed)
    Abstract [en]

    The effective anti-malarial medicine artemisinin is costly because of the low content in Artemisia annua. Genetic engineering of A. annua is one of the most promising approaches to improve the yield of artemisinin. In this work, the transcription factor AaWRKY1, which is thought to be involved in the regulation of artemisinin biosynthesis, was cloned from A. annua var. Chongqing and overexpressed using the CaMV35S promoter or the trichome-specific CYP71AV1 promoter in stably transformed A. annua plants. The transcript level of AaWRKY1 was increased more than one hundred times under the CaMV35S promoter and about 40 times under the CYP71AV1 promoter. The overexpressed AaWRKY1 activated the transcription of CYP71AV1 and moreover the trichome-specific overexpression of AaWRKY1 improved the transcription of CYP71AV1 much more effectively than the constitutive overexpression of AaWRKY1, i.e. up to 33 times as compared to the wild-type plant. However the transcription levels of FDS, ADS, and DBR2 did not change significantly in transgenic plants. The significantly up-regulated CYP71AV1 promoted artemisinin biosynthesis, i.e. up to about 1.8 times as compared to the wild-type plant. It is demonstrated that trichome-specific overexpression of AaWRKY1 can significantly activate the transcription of CYP71AV1 and the up-regulated CYP71AV1 promotes artemisinin biosynthesis.

  • 48. Heimgartner, U
    et al.
    Pietrzak, M
    Geertsen, R
    Brodelius, Peter
    Biotechnology ETH-Hönggerberg CH-8093 Zürich, Switzerland .
    da Silva Figueiredo, A C
    S Pais, Maria
    Purification and Partial Characterization of Milk Clotting Proteases from Flowers of Cynara cardunculus1990In: Phytochemistry, ISSN 0031-9422, E-ISSN 1873-3700, Vol. 29, no 5, p. 1405-1410Article in journal (Refereed)
    Abstract [en]

    Three proteases (cynarases 1, 2 and 3) with milk-clotting activity have been purified from dried flowers of Cynara cardunculus. The proteases are each composed of one large and one small subunit. The native Mr of the dimeric proteins is 49 000. The three proteases are glycoproteins containing N-linked high mannose type glycans. Cynarase 3 shows the highest proteolytic and milk-clotting activity. All three enzymes express maximum activity at pH 5.1. Inhibitor studies indicate that the cynarases are of the aspartic acid type. Antibodies raised against the large subunit of cynarase 3 cross-reacts with the large subunits of the other two cynarases after destruction of the glycan structure by periodate oxidation. 

  • 49. Hulst, A C
    et al.
    Tramper, J
    Brodelius, Peter
    Institute of Biotechnology, Swiss Federal Institute of Technology, Hönggerberg, CH-8093 Zürich, Switzerland.
    Eijkenboom, L J C
    Luyben, K Ch A M
    Immobilized Plant Cells: Respiration and Oxygen Transfer1985In: Journal of chemical technology and biotechnology (1986), ISSN 0268-2575, E-ISSN 1097-4660, Vol. 35, no 3, p. 198-204Article in journal (Refereed)
    Abstract [en]

    The influence of support material (calcium alginate, x-carrageenan and agarose), cell loading and, in case of alginate, bead diameter on the rate of respiration of immobilised plant cells (Daucus carota) was investigated. No significant differences were observed between the three supports and no loss of respiration activity occurred as result of immobilisation per se. The results show further that above a critical combination of cell loading and bead diameter limitations of the rate of respiration by diffusion of oxygen increases with increasing loading and diameter. 

  • 50. Höjeberg, B
    et al.
    Brodelius, Peter
    UNIV LUND, CTR CHEM, DEPT PURE & APPL BIOCHEM, S-22007 LUND 7, SWEDEN .
    Rydström, J
    Mosbach, K
    Affinity Chromatography and Binding Studies on Immobilized Adenosine 5'-Monophosphate and Adenosine 2'.5'-Bisphosphate of Nicotinamide Nucleotide Transhydrogenase from Pseudomonas aeruginosa1976In: European Journal of Biochemistry, Vol. 66, no 3, p. 467-475Article in journal (Refereed)
    Abstract [en]

    Nicotinamide nucleotide transhydrogenase from Pseudomonas aeruginosa was purified to apparent homogeneity with an improved method employing affinity chromatography on N6-(6-aminoyl)-adenosine-2′,5′-bisphosphate-Sepharose 4B. Polyacrylamide gel electrophoresis of the purified transhydrogenase carried out in the presence of sodium dodecyl sulphate, indicated a minimal molecular weight of 55000 ± 2000. The kinetic and regulatory properties of the purified transhydrogenase resembled those of the crude enzyme, i.e., NADPH, adenosine 2′-monophosphate and Ca2+were activators whereas NADP+was inhibitory.  Nicotinamide nucleotide-specific release of binding of the transhydrogenase to N6-(6-aminohexyl)-adenosine-2′,5′-bisphosphate-Sepharose and N6-(6-aminohexyl)-adenosine-5′-monophospliate-Sepharose suggests the presence of at least two separate binding sites for nicotinamide nucleotides, one that is specific for NADP(H) and one that binds both NAD(H) and NADP(H). Binding of transhydrogenase to N6-(6-aminohexyl)-adenosine-2′,5′-bisphosphate-Sepharose and activation of the enzyme by adenosine-2′,5′-bisphosphate showed a marked pH dependence. In contrast, inhibition of the Ca2+-activated enzyme by adenosine 2′,5′-bisphosphate was virtually constant at various pH values. This discrepancy was interpreted to indicate the existence of separate nucleotide-binding effector and active sites. 

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