PubMed Health⌕ Search

Biomedical subjects

A Hüttermann

Publications and source records attributed to A Hüttermann.

At least 19 recordsLinked to original sources

The effect of ions on the enzymatically induced synthesis of lignin graft copolymers.

The effect of different ions which are constituents of technical lignin sulfonates (LS) on chemo-enzymatic graft co-polymerization was determined. The application of the iron chelator desferrioxamine in the initial reaction mixture revealed that iron impurities of LS which catalyzed a Fenton-like reaction were crucial for the initiation of grafting, whereas calcium or chloride ions showed no such effect. The addition of laccase (ATCC 11235) to the reaction mixture which contained desferrioxamine caused a significantly higher yield compared to the control; this indicates a crucial effect of laccase with regard to the initiation of copolymerization. The involvement of laccase in the initiation of the graft copolymerization was additionally confirmed by the application of low molecular weight phenolics instead of LS. In the presence of the lignin-like substrates, 3,4-dihydroxybenzoic acid and guaiacol, the rate of the decomposition of t-butylhydroperoxide was significantly enhanced by laccase. It can be assumed that the enzymatically generated phenoxy radicals mediate the production of oxygen centered radicals (alkoxy or peroxy) which initiate grafting.

Journal Article↗

Chemo-enzymatically induced copolymerization of phenolics with acrylate compounds.

Initiation of copolymerization of lignin-like phenolic and acrylic compounds by the phenoloxidase laccase (EC 1.10.3.2) and a peroxide species (t-butylhydroperoxide, t-BHP) was compared to a Fenton-like system (ferrous ion, t-BHP). Initially, the relative activity of laccase towards different phenolic compounds and the optimum pH of some characteristic phenolics were determined. The polymer yield and the average molecular weight (Mw) of chemo-enzymatically produced polymers were dependent both on the type of each phenolic tested and on the phenol/monomer ratio. Furthermore, the success of copolymerization of the phenolics was dependent both on their redox potential and on the type of acrylic monomer applied. The extent of phenol incorporation into the polymer chain was enhanced by the presence of laccase in the reaction mixture and was significantly higher than in polymerization initiated by a Fenton-like reaction.

Acrylamide↗

Modification of lignin for the production of new compounded materials.

The cell walls of woody plants are compounded materials made by in situ polymerization of a polyphenolic matrix (lignin) into a web of fibers (cellulose), a process that is catalysed by polyphenoloxidases (laccases) or peroxidases. The first attempt to transform the basic strategy of this natural process for use in human craftsmanship was the ancient lacquer method. The sap of the lacquer tree (Rhus verniciflua) contains large amounts of a phenol (urushiol), a polysaccharide and the enzyme laccase. This oil-in-water emulsion solidifies in the presence of oxygen. The Chinese began using this phenomenon for the production of highly creative artwork more than 6,000 years ago. It was the first example of an isolated enzyme being used as a catalyst to create an artificial plastic compound. In order to apply this process to the production of products on an industrial scale, an inexpensive phenol must be used, which is transferred by an enzyme to active radicals that react with different components to form a compounded material. At present, the following approaches have been studied: (1) In situ polymerization of lignin for the production of particle boards. Adhesive cure is based on the oxidative polymerization of lignin using phenoloxidases (laccase) as radical donors. This lignin-based bio-adhesive can be applied under conventional pressing conditions. The resulting particle boards meet German performance standards. By this process, 80% of the petrochemical binders in the wood-composite industry can be replaced by materials from renewable resources. (2) Enzymatic copolymerization of lignin and alkenes. In the presence of organic hydroperoxides, laccase catalyses the reaction between lignin and olefins. Detailed studies on the reaction between lignin and acrylate monomers showed that chemo-enzymatic copolymerization offers the possibility to produce defined lignin-acrylate copolymers. The system allows control of the molecular weights of the products in a way that has not been possible with chemical catalysts. This is a novel attempt to enzymatically induce grafting of polymeric side chains onto the lignin backbone, and it enables the utilization of lignin as part of new engineering materials. (3) Enzymatic activation of the middle-lamella lignin of wood fibers for the production of wood composites. The incubation of wood fibers with a phenol oxidizing enzyme results in oxidative activation of the lignin crust on the fiber surface. When such fibers are pressed together, boards are obtained which meet the German standards for medium-density fiber boards (MDF). The fibers are bound together in a way that comes close to that by which wood fibers are bound together in naturally grown wood. This process will, for the first time, yield wood composites that are produced solely from naturally grown products without any addition of resins.

Biopolymers↗

Chemo-enzymatic synthesis and characterization of graft copolymers from lignin and acrylic compounds.

Chemo-enzymatic initiation of graft copolymerization of acrylic compounds onto different technical lignosulfonates (LS) was compared to a Fenton-like system (ferrous ion, t-BHP). The enzyme tested was a phenoloxidase laccase (EC 1.10.3.2) from the white rot basidomycete Trametes versicolor. Most applied lignins were successfully grafted, resulting in a polymer yield of more than 90%. The effect of initiator concentration and the lignin/monomer ratio on the yield and M(w) of enzymatically grafted polymers were studied. The homopolymer proportion in the enzymatically produced grafts of Ca-LS and acrylic acid was 5 to 6x lower than those initiated by the Fenton-like reagent; no such differences were observed for Na-LS.

Journal Article↗

Chemoenzymatical grafting of acrylamide onto lignin.

Laccase (E.C.1.10.3.2.) from white-rot basidiomycete Trametes versicolor and dioxane peroxides were essential in the copolymerization of acrylamide and a derivative with lignin in a dioxane-H(2)O (7:3) mixture. Both a solubility test and an elemental analysis of the eluted and separated fractions provided evidence of grafting. Gel permeation chromatography on Sephadex G-100 showed that the side chain and the lignin backbone migrate as one unit through the column. Copolymers obtained were characterized by UV-VIS-spectroscopy as well as FT-IR, and 13C-CPMAS-NMR spectroscopy. Freeze-dried copolymers of lignin and acrylamide appeared as homogeneous fibril-like particulate. The mechanism of the enzymatical grafting is discussed.

Acrylamide↗

Enhanced stability of laccase in the presence of phenolic compounds.

The storage stability of laccase (EC 1.10.3.2) from the white-rot basidomycete Trametes versicolor in potassium-citrate buffer was enhanced by various phenolic compounds as well as by lignin sulfonate. The highest storage stability was obtained with phenolics, e.g. phloroglucin and 3,5-dihydroxybenzoic acid; these represent substrates of laccase which are oxidized slowly because of their relatively high redox potential and which did not precipitate from the solution within the tested period of time. Sterilization enhanced the stability of laccase but additional stabilization by phenolics was observed both under sterile and non-sterile conditions. We thus concluded that stabilization occurred not only through prevention of microbial degradation.

Catechols↗

Genotypic variation in drought tolerance of poplar in relation to abscisic acid.

We investigated effects of water stress and external abscisic acid (ABA) supply on shoot growth, stomatal conductance and water status in 1-year-old cuttings of a drought-sensitive poplar genotype Populus x euramericana cv. I-214 (Italica) and a drought-tolerant genotype P. 'popularis 35-44' (popularis). Populus popularis was more productive and maintained higher leaf water potentials throughout the drought treatment than cv. Italica. Supply of ABA to the xylem sap caused a greater decline in growth and more leaf abscission in shoots of cv. Italica than in shoots of P. popularis. Immediately after initiation of the drought treatment in P. popularis, the ABA concentration ([ABA]) of the xylem increased rapidly and stomatal conductance declined; however, stomatal conductance had returned to control values by the third day of the drought treatment, coincident with a gradual decline in xylem [ABA]. In contrast, xylem [ABA] of cv. Italica initially increased more slowly than that of P. popularis in response to the drought treatment, but the increase continued for 3 days at which time a tenfold increase in xylem [ABA] was observed that was followed by abscission of more than 40% of the leaves. We conclude that sensitivity of poplar roots to variation in soil water content varies by clone and that a rapid short-term accumulation of ABA in shoots in response to water stress may contribute to drought tolerance.

Journal Article↗

Degradation of anthracene by laccase of Trametes versicolor in the presence of different mediator compounds.

Laccase of Trametes versicolor was generally able to oxidize anthracene in vitro. After 72 h incubation about 35% of the anthracene was transformed stoichiometrically to 9,10-anthraquinone. Transformation of anthracene increased rapidly in the presence of different mediators that readily generate stable radicals: 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 1-hydroxybenzotriazole. For the reaction, the presence of both the laccase and the mediator was necessary. In the presence of 0.005 mM 1-hydroxybenzotriazole this conversion had removed 47% of the anthracene after 72 h; 75% of the substrate was oxidized during this period when ABTS (1 mM) was used as mediator. In contrast to reactions without or with only low concentrations of a mediator, there was a discrepancy between the disappearance of anthracene and the formation of 9,10-anthraquinone in mediator-forced reactions. Coupling-products of mediators with anthracene degradation products were found. Anthracene disappeared nearly completely after incubation for 72 h with laccase in a 0.1 mM solution of 1-hydroxybenzotriazole and was transformed to 9,10-anthraquinone in about 80% yield; 90% of the substrate was transformed in the presence of ABTS (2.0 mM) resulting again in 80% quinone. Phenothiazine was not effective in this system.

Anthracenes↗

Aluminum toxicity and forest decline.

The rate of root elongation in seedlings of Picea abies was drastically inhibited by exposure to AlCl(3) at 800 or 1200 mumol/dm(3) in nutrient solutions. A 35-day exposure to Al at 700 mumol/dm(3) reduced Mg and Ca in roots and needles of the seedlings. The Mg content of needles was reduced to levels considered to be critical for Mg deficiency. In investigations of (45)Ca uptake into roots, exposure to Al at 100-800 mumol/dm(3) reduced (45)Ca uptake by 77-92%, respectively. By using x-ray microanalysis, the distribution of Al, Mg, Ca, and K was found to be similar in roots of Picea abies seedlings grown in solution culture and in roots collected from declining spruce stands at Solling, F.R.G. In solution culture Al displaced Mg and Ca in the root cortex. A mechanism of Al toxicity for root growth and ion uptake is proposed, and its relevance to forest decline is discussed.

Journal Article↗

Catabolism of single ring aromatic acids by four Aspergillus species.

Four species of the genus Aspergillus, viz A. fumigatus, A. japonicus, A. niger and A. terreus, decarboxylated, demethoxylated and ring-cleaved aromatic compounds but to different extents. Decarboxylation of vanillate occurred before ring-cleavage, which preceded the release of 14CO2 from the methoxyl group. A large proportion of labelled carbon from the ring of ferulate and vanillate was found in particulate or trichloracetic acid precipitable material of homogenized fungal mycelium. The four Aspergillus species contained vanillate-inducible protocatechuate-3,4-dioxygenase and catechol-1,2-dioxygenase activities.

Aspergillus fumigatus↗

A method for separation of pigments from plasmodia of the true slime molds, Physarum polycephalum and Physarum nudum.

Plasmodia of the true slime molds Physarum polycephalum and Physarum nudum were grown on agar semidefined medium (J. W. Daniel and H. Baldwin (1964) in Methods in Cell Physiology (Precsott, D. M., Ed.), Vol. 1, pp. 9-44, Academic Press, New York.) and pigments were extracted from 8-day-old plasmodia with the same solvent mixture that was used later as a mobile phase in the HPLC analysis. The separation was carried out on a strong anion exchanger column in a methanol/borate buffer solution. Under these conditions eight pigments from P. polycephalum and eight from P. nudum were revealed. This method can be used for detailed studies of plasmodial pigments in these species.

Chromatography, High Pressure Liquid↗

Plasmodium formation in Physarum polycephalum: cytological events and their timing relative to commitment.

Amoebae of "selfing" strains of Physarum polycephalum transform into multinucleate plasmodia without change in ploidy. The transformation occurs when cultures of amoebae reach sufficiently high cell densities or when cultures of low densities are exposed through filters to dense "inducing" cultures of amoebae. We have induced selfing amoebae to transform individually and in populations in order to study cytological changes and their timing relative to the irreversible commitment of cells to plasmodium formation. An increase in cell size was observed to precede commitment, and the first nuclear mitosis, which gives rise to a binucleate cell, was found to follow commitment.

Cell Communication↗

Aminopeptidases in seeds of picea abies (L.) Karst.: characterization of leucine aminopeptidase by molecular properties and inhibitors.

By either acrylamide or starch gel electrophoresis of Norway spruce (Picea abies) seed extracts, two prominent isoenzyme bands were obtained after staining for leucine aminopeptidase (LAP). These bands were proved to correspond to each other by reelectrophoresis in both gel media. Single endosperm studies with acrylamide gels showed clearly that, in addition to LAP, two bands are expressed after staining for alanine aminopeptidase (AAP) as well. Both the LAP and the AAP activities appeared together as a single peak between catalase and ferritin after gel chromatograhy on Sepharose. Isoelectric focusing in sucrose gradients proved the two LAP activities to have identical isoelectric points revealed that LAP, but not AAP, is detectable by standard starch gel electrophoretic procedures. The two LAP bands refer to approximate molecular weights of 71,000 and 131,000, respectively. Disaggregation studies did not conclusively determine whether these two bands represent different enzymes or not. only inhibitors succeeded in producing a definite differentiation by selective inhibition of one of the two bands. It is concluded that in both gel media the isoenzyme bands reflect the activity of two distinct leucine aminopeptidases.

Aminopeptidases↗

Muliple sites of action of cycloheximide in addition to inhibition of protein synthesis in Physarum polycephalum.

The specificity action of cycloheximide was tested using a cycloheximide resistant mutant of Physarum polycephalum. This resistance has previously been shown to reside with the ribosomes, making cytoplasmic protein synthesis refractile to the action of the drug. We show here that cycloheximide in the mutant strain causes specific alterations in metabolism without influencing the growth rate. These are: 1. lowered specific activity of glutamate dehydrogenase during starvation, 2. alteration of the molecular weight of glutamate dehydrogenase, 3. inhibition of uptake of amino acids from the medium into the internal pools. Possible explanations for these effects of cycloheximide outside of protein synthesis per se are considered. We conclude that cycloheximide may not be considered a specific inhibitor of protein synthesis, and that a causal relationship between protein synthesis and any biological process cannot be claimed unless such specificity is demonstrated in each case, preferably by use of mutants.

Amino Acids↗

Characterization and comparison of the DNAs of the three closely related bacteriophages gd, ge and gf with the genome DNA of the hydrogen-oxidizing host strain Pseudomonas pseudoflava GA3.

The double-stranded (ds)DNAs of the three closely related temperate Pseudomonas pseudoflava bacteriophages gd, ge and gf were studied biochemically and biophysically. The GC content of the DNA was 67.4 +/- 0.5% and differed only slightly from that of the host P. pseudoflava. By electron microscopic length measurements a mol. wt. of 26.1 X 10(6) to 26.7 X 10(6) was calculated for the three bacteriophage DNAs. Homogeneity of the bacteriophage DNAs was further demonstrated by specific cleavage with restriction endonucleases WcoRI and HindIII. It was concluded that the three homo-immune bacteriophages are identical. The genome size of the host P. pseudoflava GA3 was 3.7 X 10(9) as calculated from optical renaturation rate measurement with Xanthomonas pelargonii reference DNA. The bacteriophage gd genome thus amounts to 0.7% of the chromosome of this bacterial host.

Bacteriophages↗

Melanin biosynthesis during differentiation of Physarum polycephalum.

Melanin synthesis in the myxomycete Physarum polycephalum occurs during sporulation but not during spherule formation. Melanin-like pigment was extracted from spores. An almost identical substance of polyphenols was extracted from spherules and characterized by its ultraviolet and infrared absorbance spectra. Polyphenol oxidase activity in spherules was very low and showed only one weak isoenzyme band in isoelectric focusing polyacrylamide gels. A much higher activity, and an increasing number of isoenzymes, were detected in sporulating cultures after illumination during the differentiation process. The addition of melanin precursors resulted in the synthesis of brownish-yellow spherules, probably containing dopachrome, whereas the addition of polyphenol oxidase inhibitors resulted in yellow sporangia. The results indicate that melanin synthesis is probably only a stage in maturation but not an essential part of the morphogenetic process itself.

Catechol Oxidase↗

Studies on the cell cycle of Myxobacter AL-1. II. Activities of seven enzymes during the cell cycle.

The properties of seven enzymes were studied in extracts from Myxobacter AL-1. The enzymes were isocitrate dehydrogenase (E.C.1.1.1.42), succinate dehydrogenase (E.C.1.3.99.1), alkaline phosphatase (E.C.3.1.3.1), alpha-glucosidase (E.C.3.2.1.20), beta-glucosidase (E.C.3.2.1.21), beta-galactosidase (E.C.3.2.1.23), and N-acetyl-glucosaminidase (E.C. 3.2.1.30). Four of these enzymes: isocitrate dehydrogenase, alpha-glucosidase, beta-glucosidase, and beta-galactosidase are cytosolic enzymes. Succinate dehydrogenase was found to be located on the cytoplasmic membrane system, whereas alkaline phosphatase and N-acetylglucosaminidase were considered as enzymes which bind the outer membranes resp. the cell wall. During the cell cycle, all enzymes have a pattern of discontinuous activity increase. Succinate dehydrogenase and isocitrate dehydrogenase exhibit a stepwise increase of activity, whereas the other enzymes follow the pattern of a peak enzyme.

Acetylglucosaminidase↗