Starch digestion and adsorption by beta-amylase of Emericella nidulans (Aspergillus nidulans).

A mutant strain of Emericella nidulans MNU 82 was isolated by multistep mutation. The beta-amylase produced by the mutant was able to digest raw starch. It was readily and strongly adsorbed onto raw starch at pH 5.0. The enzyme to starch ratio was 1950 U/g starch. The enzyme showed no correlation between the capacity of raw starch digestion and adsorption of the enzyme.     

Starch digestion and adsorption by β-amylase of Emericella nidulans (Aspergillus nidulans)

B. CHATTERJEE, A. GHOSH AND A. DAS. 1992. A mutant strain of Emericella nidulans MNU 82 was isolated by multistep mutation. The β-amylase produced by the mutant was able to digest raw starch. It was readily and strongly adsorbed onto raw starch at pH 5.0. The enzyme to starch ratio was 1950 U/g starch. The enzyme showed no correlation between the capacity of raw starch digestion and adsorption of the enzyme.     

Production of xylanase by Emericella nidulans NK-62 on low-value lignocellulosic substrates

Thermotolerant Emericella nidulans NK-62 was isolated from bird nesting material and was tested for its ability to produce xylanase. The fungus when grown on a medium containing wheat bran (2% w/v) supplemented with Czapek's mineral salt solution at 45 °C for 7 days produced 362 IU/ml of xylanase (EC 3.2.1.8). The specific activity of E. nidulans NK-62 xylanase was found to be 275 IU/mg of total protein. The enzyme was found to be active over a broad temperature and pH range with 60 °C as optimum temperature for enzyme activity. The enzyme was stable at 50 °C and its half-life at 55 °C was 45 min. -xylosidase (EC 3.2.1.37) and carboxymethylcellulase (EC 3.2.1.4) activities, 0.018 and 0.21 IU/ml respectively, were also noticed. The fungus was screened for its ability to produce xylanase on four different lignocellulosic substrates. It produced 318.9 IU/ml of cellulase-free xylanase on corn cobs. The fungus could also utilize lentil bran (seed husk of Lens esculentus) and meal of groundnut shells to produce 84.8 and 17.3 IU/ml xylanase respectively.     

Microsatellite markers for assessing DNA polymorphism of Emericella nidulans in nature

Based on genome sequence of Emericella nidulans, primers for 25 microsatellites were designed, of which, seven were chosen as polymorphic markers (five to 12 alleles). Simple sequence repeat (SSR) polymorphism was tested in 44 isolates from extremely arid soils of the southern Negev desert, Israel and some related species of Aspergillus. The SSR marker set appears suitable for population studies in diverse ecological conditions.     

Confirmation of the relationships of Aspergillus egyptiacus and Emericella nidulans using progesterone transformation

Isolates of Emericella nidulans (7 isolates), Aspergillus egyptiacus (7) and A. versicolor (5) were tested for their ability to transform progesterone. Consistent results were obtained between isolates of the same species. Whilst A. egyptiacus and E. nidulans possessed the enzyme system catalysing the transformation of progesterone into 11-α-hydroxyprogesterone, A. versicolor did not. Thus, progesterone transformation could be considered as further diagnostic evidence that A. egyptiacus is related to the Emericella group.     

Inheritance of guttural pouch tympany in the arabian horse

The objective of the present study was to analyze the mode of inheritance of guttural pouch tympany (GPT) using pedigrees of Arabian horses. Complex segregation analyses were employed to test for the significance of nongenetic transmission and for monogenic, polygenic, and mixed monogenic-polygenic modes of inheritance. Horses affected by GPT comprised 27 Arabian purebred foals. Of these 27 animals, 22 were patients at the Clinic for Horses, School of Veterinary Medicine Hannover, Hannover, Germany, between 1994 and 2001 and 5 Arabian foals were from stud farms. Information on the pedigrees of these patients allowed us to classify the affected foals into four families with a total of 276 animals. The regressive logistic model analysis took into account the nonrandomness of the pedigrees through multiple single ascertainment correction. The complex segregation analysis showed that, among all other models employed, a polygenic and a mixed monogenic-polygenic model best explained the segregation of Arabian foals with GPT. Models including only nongenetic distributions and monogenic inheritance could be significantly rejected. This is the first report in which a genetic component could be shown to be responsible for GPT in horses.     

Effect of pH on the stability of Pleurotus eryngii versatile peroxidase during heterologous production in Emericella nidulans

Complementary DNA (cDNA) encoding the new versatile peroxidase from the ligninolytic basidiomycete Pleurotus eryngii has been expressed in the ascomycete Emericella nidulans. In recombinant E. nidulans cultures, the pH reached values as high as 8.3, correlating with a sharp decrease in peroxidase activity. Peroxidase was rapidly inactivated at alkaline pH, but was comparatively stable at acidic pH. The peroxidase inactivation in alkaline buffer could be reversed by adding Ca²⁺ and lowering the pH. However, reactivation did not result after incubating the enzyme in non-buffered E. nidulans cultures that reached pH 7.5. To optimize recombinant peroxidase production, the effect of controlling the pH in E. nidulans bioreactor cultures was studied. An extended growth period, and a significant increase in the recombinant peroxidase level (5.3-fold higher activity than in the bioreactor without pH control) was obtained when the pH was maintained at 6.8, showing that culture pH is an important parameter for recombinant peroxidase production.     

Small enzymes with esterase activities from two thermophilic fungi, Emericella nidulans and Talaromyces emersonii

Extracellular esterase activities in Emericella nidulans and Talaromyces emersonii are attributed to small enzymes with molecular weights less than 10 kDa (microenzymes). A 1.6 kDa esterase accounted for most of the esterase activity observed in both organisms and one of them also contained a 4.1 kDa microenzyme with weaker esterase activity. These esterases were growth-associated and active towards fluorescein dibutyrate and -naphthyl acetate as well as tributyrin.     

Diversity of microsatellites in natural populations of ascomycetous fungus, Emericella nidulans, in Israel on local and regional scales

The genetic divergence of Israeli populations of soil ascomycetous fungus Emericella nidulans was studied on regional and local scales using seven microsatellite (SSR) trinucleotide markers. The study was performed in the framework of the “Evolution Canyon” research program at the Institute of Evolution, University of Haifa, in three “Evolution Canyons” (EC): EC I (Mt. Carmel), EC II (western Upper Galilee), and EC III (the southern Negev desert). The first two canyons (EC I and EC II) are located in the northern part of Israel at a distance of 38 km. EC III is located to the south at a distance of nearly 350 km from the northern ECs. In each canyon, E. nidulans strains were isolated from the opposite slopes and (in EC III) the valley bottom. All three EC populations of E. nidulans were found to be genetically distinct. The estimated genetic divergences correspond to geographical distances and ecological differences among the three studied microsites. On a regional scale, SSR polymorphism tends to increase with severity of ecological conditions. In general, the effect of “structural” factors (predicted number of repeats in SSR markers and distance to the centromere) in the microsatellite diversity was higher compared to ecological factors.     

Emericella appendiculata, a new species from Chinese soil

Emericella appendiculata, a new species isolated from soil of the Pamire Plateau, is described and illustrated. It is characterized by grayish green non-ostiolate ascomata surrounded by a thick layer of hülle cells, membranaceous peridium, prototunicate asci, violet-brown, lenticular ascospores which are ornamented by two stellate equatorial crests, capitate convex surfaces, and long filiform appendages, and anAspergillus anamorph with biseriate conidiogenous cells.     

Emericella omanensis,a new species from Oman soil

A new species ofEmericella isolated from forest soil in the Oman,E. omanensis, is described and illustrated. It differs from the other known species of the genus in having bivalvate ascospores with a tuberculate or verruculose convex wall. The new species is compared with the closely related speciesE. desertorum andE. echinulata.     

Emericella qinqixianii, a new species from desert soil in China

Emericella qinqixianii, a new species isolated from desert soil from Sanchakou, Aksu, Qiemo, Yuli, Yutian, and the Taklimakan desert 100 km inland from Minfeng, Xinjiang Province, China, is described and illustrated. It is characterized by grayish yellow to olive brown, non-ostiolate ascomata surrounded by hyaline to pale yellowish brown hülle cells, membranaceous peridium, prototunicate asci, and violet-brown, lenticular ascospores with two equatorial crests, smooth convex surfaces, and long filiform appendages. It hasAspergillus anamorph with biseriate aspergilla.     

Guttural pouches, brain temperature and exercise in horses

Selective brain cooling (SBC) is defined as the lowering of brain temperature below arterial blood temperature. Artiodactyls employ a carotid rete, an anatomical heat exchanger, to cool arterial blood shortly before it enters the brain. The survival advantage of this anatomy traditionally is believed to be a protection of brain tissue from heat injury, especially during exercise. Perissodactyls such as horses do not possess a carotid rete, and it has been proposed that their guttural pouches serve the heat-exchange function of the carotid rete by cooling the blood that traverses them, thus protecting the brain from heat injury. We have tested this proposal by measuring brain and carotid artery temperature simultaneously in free-living horses. We found that despite evidence of cranial cooling, brain temperature increased by about 2.5 degrees C during exercise, and consistently exceeded carotid temperature by 0.2-0.5 degrees C. We conclude that cerebral blood flow removes heat from the brain by convection, but since SBC does not occur in horses, the guttural pouches are not surrogate carotid retes.     

Temperature dependence of horse heart cytochromec oxidation by membrane preparations ofAnacystis nidulans: Characterization of two distinct arrhenius discontinuities

The oxidation of reduced horse heart cytochromec by membranes isolated from the cyanobacteriumAnacystis nidulans after growth at different temperatures was studied between 4°C and 41°C in the light and the dark using both spectrophotometric and polarographic techniques. Arrhenius plots of the temperature dependence of cytochromec photooxidation showed a single discontinuity at 25°C, 15°C, and 12°C in membranes derived from cells grown at 40°C, 30°C, and 25°C, respectively. By contrast. Arrhenius plots of the temperature dependence of dark respiratory cytochromec oxidation always displayed two distinct breaks at 25 and 18°C, 15 and 8.5°C, and 12 and 5.5°C in membranes isolated from cells grown at 40°C, 30°C, and 25°C, respectively. The results are discussed in terms of the thermotropic lipid-phase transitions known to take place in the membranes ofA. nidulans. Special reference will be made to possibly distinct localizations of the membrane-bound cytochromec oxidase complexes in respiration and photosynthesis.     

Ringworm infection by Microsporon canis in a horse

Summary The authors comment on horses ringworm byM. canis. They present a clinical, mycological and epidemiological observation on the first Brazilian case.

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Sumário Os autores comentam a incidência de Tinha porM. canis em cavalos e sua raridade atualmente. Apresentam a observacão clínica, micológica e epidemiológica do primeiro caso brasileiro de Tinha em equino devida aoM. canis.

     

Transformation by integration in Aspergillus nidulans

DNA-mediated genetic transformation of Aspergillus nidulans has been achieved by incubating protoplasts from a strain of A. nidulans carrying a deletion in the acetamidase structural gene with DNA of derivatives of plasmid pBR322 containing the cloned structural gene for acetamidase [Hynes et al., Mol. Cell. Biol. 3 (1983) 1430-1439; p3SR2] in the presence of polyethylene glycol and CaCl2. The highest frequency obtained was 25 transformants per microgram of DNA. No enhancement of the transformation frequency was observed when DNAs of plasmids carrying either a fragment of the A. nidulans ribosomal repeat (p3SR2rr) or a fragment containing a possible A. nidulans mitochondrial origin of replication (p3SR2mo) in addition to the acetamidase gene were used. Both pBR322 and acetamidase gene sequences become integrated into the genome of A. nidulans in transformant strains. Integration events into the residual sequences adjacent to the deletion in the acetamidase gene, and probably (for p3SR2rr and p3SR2mo) into the ribosomal repeat unit are described.     

Experimental acute poisoning in mice induced by emestrin,a new mycotoxin isolated from Emericella species

The effects of emestrin (EMS), a secondary metabolite of the Emericella species, on male ICR mice were examined. The intraperitoneal LD₅₀ values of EMS were 17.7 and 13.0 mg/kg at 24 and 48 hr, respectively. The target organs of EMS were the heart, liver and thymus. In doses over 30 mg/kg the experimental animals died from cardiac failure shortly after the injections. Several survivors that were given EMS in doses under 20 mg/kg showed severe centrilobular necrosis in the liver at 24 hr. Marked degeneration of mitochondria was seen in electron micrographs of both cardiac muscle cells and hepatocytes. In the degenerated hepatocytes, prominent proliferation of RER, membrane-limited inclusions containing both ribosome-like granules and RER, and fenestrated lamella-like structures were observed. Massive necrosis of lymphocytes was always observed in the cortical layer of the thymus of the survivors within 24 hr, while bilateral adrenalectomized mice showed no discernible pathomorphological changes in the lymphoid tissues. Pretreatment of mice with diethyl maleate increased the incidence and severity of hepatic necrosis, whereas that with either cysteine or CoCl₂ reduced the severity of centrilobular necrosis of the liver. Pretreatment with phenobarbital had no significant effect on EMS-induced hepatic lesions.