Glycogen Metabolism in Aureobasidium Pullulans: A Glycogen Synthetase with Unusual Activation Properties

Abstract

Cultures of filamentous fungi that secrete significant amounts of exopolysaccharides are among the most difficult of fermentation fluids, presenting difficulties in the areas of aeration, agitation, mixing, and control that may in turn impact the physiology of the microorganism in an undesirable manner. The fungus Sclerotium glucanicum, which produces a potentially useful exopolysaccharide known as scleroglucan, illustrates many such difficulties. This review discusses in detail the range of physiological studies on the producing microorganism itself, including those concerning formation of “undesirable” byproducts, principally oxalate, but also, under certain conditions, other TCA cycle acids. In addition, the bioreactor technology in use for production of this type of biopolymer is discussed in relation to the difficulties such fluid types present. The potential of pneumatically agitated reactors for such production is evaluated, and the lack of fundamental studies on such reactors and on the hydrodynamics and mixing behavior of such complex fluids is pointed out.     

Senescence in cut carnation flowers: Temporal and physiological relationships among water status, ethylene, abscisic acid and membrane permeability

Changes in water status, membrane permeability, ethylene production and levels of abscisic acid (ABA) were measured during senescence of cut carnation flowers ( Dianthus caryophyllus L. cv. White Sim) in order to clarify the temporal sequence of physiological events during this post-harvest period. Ethylene production and ABA content of the petal tissue rose essentially in parallel during natural senescence and after treatment of young flowers with exogenous ethylene, indicating that their syntheses are not widely separated in time. However, solute leakage, reflecting membrane deterioration, was apparent well before the natural rise in ethylene and ABA had begun. In addition, there were marked changes in water status of the tissue, including losses in water potential (ψ_w), and turgor (ψ_p), that preceded the rise in ABA and ethylene. As senescence progressed, ψ_w continued to decline, but ψ_p returned to normal levels. These temporal relationships were less well resolved when senescence of young flowers was induced by treatment with ethylene, presumably because the time-scale had been shortened. Thus changes in membrane permeability and an associated water stress in petal tissue appear to be earlier symptoms of flower senescence than the rises in ABA or ethylene. These observations support the contention that the climacteric-like rise in ethylene production is not the initial or primary event of senescence and that the rise in ABA titre may simply be a response to changes in water status.     

Structure of the abdominal receptor responsive to internally applied pressure in the blood-feeding insect,Rhodnius prolixus

Summary The sensory receptor responsive to pressure applied internally to the ventral abdominal body wall of the blood-feeding insects, Rhodnius prolixus, is a single sense cell containing, at its distal end, a cilium enclosed within a scolopale, a densely staining structure characteristic of insect scolopidial sensilla. A small spherical structure lies within a dilation near the midregion of the cilium, and contains nine heavily staining bodies, the position of each corresponding to a pair of microtubules in the cilium. Proximal to the dilation, the microtubules are organized in a ring of nine pairs with one microtubule of each pair associated with dyneinlike arms. Dastal to the dilation a central pair of microtubules is present, but dyneinlike arms are absent. The scolopale cell, which gives risc to the scolopale, has cytoplasmic invaginations that form an elaborate array of extracellular compartments surrounding the body wall of the sense cell. These compartments may serve to dampen high frequency vibrations permitting the receptor to respond to pressure exerted by touch, an attribute in keeping with the receptor's proposed function of detecting abdominal distension related to the size and movement of the stomach.     

Active human erythropoietin expressed in insect cells using a baculovirus vector: A role for N-linked oligosaccharide

Biologically active recombinant human erythropoietin has been expressed at high levels in an insect cell background. Expression involved the preparation of a human erythropoietin cDNA, the transfer of this cDNA to the Autographa californica nuclear polyhedrosis virus (AcNPV) genome under the polyhedrin gene promoter, and the subsequent infection of Spodoptera frugiperda cells with recombinant AcNPV. Erythropoietin cDNA was prepared through the expression of the human erythropoietin gene in COS cells using pSV2 and the construction of a COS cell cDNA library in bacteriophage Lambda GT10. Prior to transfer to the AcNPV genome, erythropoietin cDNA isolated from this library was modified at the 3′-terminus in order to replace genomic erythropoietin for SV40 cDNA derived from pSV2. Transfer of this cDNA to AcNPV and the infection of S. frugiperda cells with cloned recombinant virus led to the secretion of erythropoietin: based on bioassay, rates of hormone secretion (over 40 U/ml per h) were 50-fold greater than observed for COS cells. The purified recombinant product possessed full biological activity (at least 200000 U/mg), but was of lower M_r (23000) than human erythropoietin produced in COS cells (30000) or purified from urine (30000 to 38000). This difference was attributed to the glycosylation of erythropoietin in S. frugiperda cells with oligosaccharides of only limited size. Further removal of N-linked oligosac-charides from this M_r 23000 hormone using N-Glycanase yielded an apo-erythropoietin (M_r 18000) which possessed substantially reduced biological activity. These results indicate that glycosylation, but not the normal processing of oligosaccharides to complex types, is required for the full hormonal activity of human erythropoietin during red cell development.     

PHYSIOLOGICAL MODIFICATIONS RELATED TO DENSITY INCREASE IN PERIPHYTIC ASSEMBLAGES 1

Periphytic communities in running waters were examined as they developed on granite rocks, concrete balls and glass slides. At equivalent cell densities, no differences in pigment concentrations, species diversity or production levels were found among the different substrata examined. Development of the assemblage appeared to result from the elongation of short algal filaments which had initially settled on the surface. As these communities matured, a distinct canopy and understory developed. Cellular metabolisms were comparable among the communities. In the understory of the communities, even though the cellular content of chl a and b did not differ, chl c and carotenoid pigment concentrations were higher than those in the over-story. Bicarbonate assimilation of Tabellaria fenestrata (Lyng.) Külz. and Eunotia pectinalisi var. pectinalis (O. F. Müll?) Rabh. was higher than that of the more abundant Tabellaria flocculosa (Roth.)Kütz. var. flocculosa IV (sensu Koppen) at both high and low cell densities. This probably reflects a seasonal succession of colonizing species. Glucose assimilation appeared to be mainly attributable to bacterial activity, and algal cells of the upper layer were less active than those of the bottom. The small amount of glucose that was incorporated by the algal cells was probably absorbed passively since its amount was in direct proportion to cell volumes.     

Constancy of ascending projections in the metamorphosing brain of the meal-beetle Tenebrio molitor L. (Insecta: Coleoptera)

Summary The fate of ascending projections of thoracic interneurons in the metamorphosing brain of Tenebrio molitor is described. Persistent brain neurons were identified and their fate is described during metamorphosis. The projection sites of ascending elements are invariable throughout metamorphosis both in quantitative and in qualitative terms. Some of these ascending neurons are serotonin-immunoreactive and this set of neurons maintains a constant projection site within the metamorphosing brain. The alterations in the projection sites of these and other ascending neurons in the ventral nerve cord were analysed experimentally. The central projection sites of these persistent ascending neurons are not important for the maintenance of their nerve cord projections throughout metamorphosis. Experimental deletion of ascending neurons which project into the suboesophageal ganglion varies the shape of persistent central neurons.     

The midline metathoracic ear of the praying mantis,Mantis religiosa

Summary The praying mantis, Mantis religiosa, is unique in possessing a single, tympanal auditory organ located in the ventral midline of its body between the metathoracic coxae. The ear is in a deep groove and consists of two tympana facing each other and backed by large air sacs. Neural transduction takes place in a structure at the anterior end of the groove. This tympanal organ contains 32 chordotonal sensilla organized into three groups, two of which are 180° out of line with the one attaching directly to the tympanum. Innervation is provided by Nerve root 7 from the metathoracic ganglion. Cobalt backfills show that the auditory neuropile is a series of finger-like projections terminating ipsilaterally near the midline, primarily near DC III and SMC. The auditory neuropile thus differs from the pattern common to all other insects previously studied.     

Evidence and bioassay for diuretic factors in the nervous system of larvalHeliothis virescens

Summary Diuretic factors were studied in the central nervous system of larvae of the tobacco budworm,Heliothis virescens, using [¹⁴C]urea as a sensitive indicator for water movement through isolated Malpighian tubules. The assay required Na⁺ and a pH of 6.0–6.2 for maximum activity. Malpighian tubules had high secretory activity in feeding larvae of the fifth instar, but the activity declined during the burrowing-digging stage that preceded pupation. Malpighian tubules from starved larvae showed a greater response to extracts of nervous tissues than did tubules from feeding larvae, and extracts showed a dose-response relationship with fluid secretion. Diuretic activity was distributed throughout all parts of the central nervous system with the brain having the most activity. Brain extracts increased fluid secretion by in vitro Malpighian tubules by more than 3-fold and doubled the rate of dye clearance from the hemolymph in vivo. Diuretic activity in nervous tissue extracts was unaffected by boiling but sensitive to proteases. Fluid secretion by in vitro tubules was increased by cAMP, dbcAMP, theophylline, octopamine and dopa. These studies provide evidence for the presence of diuretic factors in the central nervous system ofH. virescens larvae and describe a sensitive bioassay for these factors.Abbreviations AR activation ratio - cAMP cyclic AMP - dbcAMP dibutyryl cyclic AMP - dbcGMP dibutyryl cyclic GMP - Dopa dihydroxyphenylalanine - 5-HT 5-hydroxytryptamine - L1 larval instar - VCNS ventral central nervous system     

Neural activity pattern is not necessary for the development of adult ultrastructure in katydid (Neoconocephalus robustus) singing muscles

Summary The singing muscles of the katydid Neoconocephalus robustus develop adult ultrastructure late in the last nymphal instar and during the first few days of adult life. The ultrastructural changes during early adulthood were not affected by unilateral axotomy shortly after the adult molt. Both denervated and innervated muscles developed adult proportions of mitochondria, myofibril, and sarcoplasmic reticulum and transverse tubules.     

Thermoanaerobium lactoethylicum spec. nov. a new anaerobic bacterium from a hot spring of Kamchatka

A new anaerobic thermophilic Gram-positive, nonsporeforming bacterium strain ZE-1 was isolated from a hot spring of Kamchatka (USSR). The cells are rod-shaped, (0.5–0.8 · 2.0–20 m), non-motile. The bacterium can grow between 42 and 75°C; the optimal temperature is 65°C. The growth is possible between pH values 5.0 and 8.5; optimal pH is 7.0. The cultures grow on the media containing peptone, yeast extract, or casein hydrolysate as nitrogen sources in the presence of glucose or some other sugars, mannitol or starch. The main fermentation products of glucose are ethanol, acetate, lactate, H₂, CO₂; byproducts are propionic, butyric and isovaleric acids. Glucose is metabolized via Embden-Meyerhoff-Parnas pathway. Molecular hydrogen does not inhibit growth. The bacterium does not reduce aceton to isopropanol, but is able to form H₂S from elemental sulfur. The bacterium contains a soluble hydrogenase. This enzyme catalyzes both evolution and uptake of H₂ and is active in the presence of methyl viologen. The DNA-base composition is 34.6 mol%; the genome size 2.08x10⁹ D. The name proposed for the isolated bacterium strain ZE-1 is Thermoanaerobium lactoethylicum spec. nov.