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J R Colvin

Publications and source records attributed to J R Colvin.

At least 37 records · Page 2Linked to original sources

Purification and properties of a soluble polymer of glucose from cultures of Acetobacter xylinum.

A soluble nondialyzable polymer of glucose was isolated and purified by selective ethanol and ammonium sulfate precipitation from the supernatant of a culture of Acetobacter xylinum which was actively producing cellulose. This polymer was heterogeneous in size with an average sedimentation constant S20,w, of the most abundant fraction of 11.1. On drying from dilute solution in water, the polymer(s) showed extended linear fibrils or aggregates of such fibrils by transmission electron microscopy. The infrared spectrum resembled closely that of cellulose II. Preparations of the lyophilized polymer were amorphous by X-ray diffraction. Composition and structure of the polymer were established by enzymatic digestion, acid hydrolysis before paper chromatography, and methylation followed by gas-liquid chromatography. Glucose was the only component of the polymer. There were few, if any, alpha 1 leads to 4 linkages between glucose residues. The polymer(s) is a linear chain of glucose units linked beta 1 leads to 4 with single glucose residues as branches at position 2 of every third glucose on the average. The possibility that this branched glucose polymer is an intermediate in cellulose biosynthesis is examined.

Chemical Phenomena↗

New evidence for an intermediate polymer of glucose in cellulose biosynthesis by Acetobacter xylinum.

The results of sucrose-density-gradient centrifugation of a cell-free particulate enzyme system from Acetobacter xylinum which was incubated with uridine diphosphoglucose indicate that there is a polymeric intermediate in the biosynthesis of cellulose. This intermediate has the properties of an oligomer of glucose, is normally attached to the heaviest particle of the suspension, but, when released by hydrolysis, is preferentially adsorbed to fragments of preformed cellulose. It may form short segments of microfibrils when precipitated from alkaline solution by ethanol. The presence of this intermediate raises again the question of a primer in cellulose biosynthesis.

Carbon Radioisotopes↗

Variability of the iron, copper and mercury contents of individual red blood cells.

The relative iron, copper and mercury contents of individual, isolated erythrocytes from eight people were determined by analytical electron microscopy. The variation in iron content between erythrocytes of the same sample is more than six times, for copper content more than tem times and for mercury more than five times. Similar variations were observed for 1-day-old chick defintive erythrocytes and for 4-day-old chick embryo primitive erythrocytes. The range of variation does not depend greatly, it at all, on the age of the erythroyctes or the tissue of origin. There is little or no correlation between the variation of iron contnet and that of copper. The cause of the wide variation of metallic ion contnt among erythroyctes is not yet known.

Adult↗

The structure of novel C35 pentacyclic terpenes from Acetobacter xylinum.

A novel C(35) terpene and its monounsaturated analogue were isolated from cultures of Acetobacter xylinum, together with traces of their C(36) homologues. These substances were found to be hopane derivatives substituted by a five-carbon chain bearing four vicinal hydroxyl groups. For the parent hydrocarbon the term bacteriohopane is proposed. The elucidation of the structures utilized high-resolution mass spectrometry of the terpenes, degradation to C(32) hydrocarbons and detailed mass-spectrometric comparison of these with C(32) hydrocarbons synthesized from known pentacyclic triterpenes. High-resolution mass-spectral data of the terpenes are presented. N.m.r. data are in agreement with the proposed structures, which are further supported by the isolation from the same organism of 22-hydroxyhopane and derivative hopene(s).

Chromatography, Gas↗

Induction of orientation of bacterial cellulose microfibrils by a novel terpenoid from Acetobacter xylinum.

1. The bacterium Acetobacter xylinum produces extracellular cellulose microfibrils that form a pellicle in the medium enmeshing the bacterial cells. These microfibrils may show some localized alignment, which can be seen as birefringence when the culture is viewed between crossed Polaroid sheets. 2. An increase in birefringence can be induced by the addition of small amounts of certain classes of lipids, particularly sterols, to the cultures. 3. A crude lipid extract from Acetobacter cells induced greatly increased birefringence when added to fresh cultures of this organism. 4. When the bacterial lipids were fractionated, most of the activity was recovered in a complex, polar lipid. The lipid is secreted into the medium during growth and is unstable. The non-saponifiable portion of this lipid is shown to be a 1:1 mixture of a saturated and a monounsaturated C(35) tetrahydroxy terpene with a hopane ring system in the accompanying paper by Förster et al. (1973). The saturated molecule is referred to as tetrahydroxybacteriohopane. 5. Tetrahydroxybacteriohopane is itself capable of inducing birefringence in cultures as is 22-hydroxyhopane, which was also isolated from the non-saponifiable fraction of the total lipids. 6. The mechanism of induction of birefringence (orientation of microfibrils) is not known. This is unlikely to be a specific effect, since all the above compounds are active (intact lipid, tetrahydroxybacteriohopane, 22-hydroxyhopane), as are other classes of lipid. It is suggested, however, that a common mechanism may be involved and that similar compounds may be concerned with control of microfibril alignment in the cells of higher plants.

Alcohols↗

Electron-opaque fibrils and granules in and between the cell walls of higher plants.

The components of higher-plant cell walls which become electron-opaque after staining with ruthenium-osmium were studied by electron microscopy. A fibrillar material which absorbs this stain is a major wall constituent in the root epidermal cells of carrot and morning glory. In both form and size, these fibrils resemble those found on the surface of suspension-cultured cells of the same species Some cells of woody species show an irregular distribution of electron-opaque material in the cell wall matrix and middle lamella. This material, which has an amorphous appearance with many electron stains, is shown by ruthenium-osmium staining to be an aggregate of discrete granules, 150-220 A in diameter. These observations are not consistent with the concept of the cell wall matrix and middle lamella as an amorphous, uniform gel

Cell Wall↗

Lignofibrils on the external cell wall surface of cultured plant cells.

Small strands and bundles of strands extend from the outside surface of suspension-cultured cells of Daucus, Ipomoea, and Phaseolus into the medium. This fibrous cell coat is present in all samples from various growth stages but appears to increase in quantity in the order Ipomoea < Phaseolus < Daucus. The bundles are often many microns in length and display great variation in frequency, size, and form. Identification of the composition of the strands and bundles as lignin is consistent with the following observations: alkaline nitrobenzene oxidation of the strands to compounds which resemble monomers of wood lignin; resistance of the strands to pronase, trypsin, pectinase, and lipase; strong irreversible adsorption of heavy metals; deposition of silver granules by treatment with silver nitrate-hexamine reagent; extraction of the bundles with aqueous dioxane (Björkman procedure); presence in quantity of a structured form of Klason lignin; and existence of material giving a positive test with the Wiesner reagent. Large individual strands (lignofibrils) from Phaseolus show the form of a flat ribbon with very thin branches at irregular intervals. This form does not vary with preparatory techniques, although its electron opacity does. Intercellular spaces display considerable structure and sometimes contain sheets of fibrillar material merging with both the middle lamella between the cells and the surface bundles facing the medium. These sheets are probably another form of association of the lignofibrils. It is suggested that natural fibrous lignin may be a much commoner component of plant tissue than suspected hitherto.

Cell Fractionation↗

Structure of the cell wall of Pythium debaryanum.

The structure of hyphal walls of Pythium debaryanum was investigated by electron microscopy of shadowed replicas and thin sections, before and after digestion by snail gut enzymes or by 1 n HCl at 100 C for 1 hr, and by X-ray diffraction. We found that the wall had two phases, one composed of microfibrils of unknown composition and a second consisting of an amorphous matrix, part of which stained like protein with potassium permanganate and part of which was removed by snail-gut enzymes. In the microfibrillar phase, there were two layers; an outer, thicker layer of randomly disposed microfibrils and an inner, thin layer of microfibrils oriented parallel to the hyphal axis. As in Neurospora crassa, the amorphous phase included a branching system of pores, 40-80 A in diameter. Unlike N. crassa, the cytoplasm of Pythium showed Golgi bodies frequently, and many lomasomes were observed between the cytoplasmic membrane and the wall. The relations between these organelles and the mechanism of wall formation in Pythium are not understood.

Cell Wall↗