Metabolism of dimethylterephthalate byAspergillus niger

Aspergillus niger (AG-1) metabolized dimethylterephthalate through monomethylterephthalate, terephthalate and protocatechuate. Degradation of dimethylterephthalate was followed by extraction of residual dimethylterephthalate from the spent medium. The quantitative UV analysis showed that 58% of the dimethylterephthalate supplement was taken up in 144 h. The metabolites were isolated from resting cell cultures. Thin layer chromatography analysis of the extract revealed the presence of two intermediates, monomethylterephthalate and terephthalate. Use of an inhibitor in resting cell culture experiment demonstrated the accumulation of protocatechuate. The time course of protocatechuate accumulation was also studied. Metabolites were identified by employing various physicochemical methods. Enzyme studies using cell-free extracts exhibited dimethylterephthalate esterase and protocatechuate dioxygenase activities. Protocatechuate was oxidized by themeta cleavage pathway. A tentative pathway for the degradation of DMTP has been proposed inA. niger.Abbreviations A. niger Aspergillus niger (AG1) - DMSO dimethyl sulfoxide - DMTP dimethylterephthalate - MMTP monomethylterephthalate - MS mass spectra - NMR nuclear magnetic resonance spectra - PCA protocatechuate - TLC thin layer chromatography - TP terephthalate - UV ultra violet spectra     

Degradation of homophthalic acid byAspergillus niger

The fungusAspergillus niger degraded homophthalic acid through the involvement ofo-hydroxyphenylacetic acid and homogentisic acid as the metabolic intermediates. Isolation of intermediates was carried out by extracting the spent medium and by using inhibitor in replacement culture techniques. Metabolites were characterized by various physicochemical methods. Oxygen uptake studies and enzyme investigations also confirmed that the degradation of homophthalic acid follows through these intermediates in the fungus.     

Characterization of phytase produced byAspergillus niger

The extracellular activity ofAspergillus niger phytase at the end of the growth phase was 132 nkat/mL in a laboratory bioreactor. The purified enzyme has molar mass approximately 100 kDa, pH optimum at 5.0, temperature optimum at 55°C and high pH and temperature stability. TheK _m for dodecasodium phytate, calcium phytate and 4-nitrophenyl phosphate are 0.44, 0.45 and 1.38 mmol/L, respectively. The enzyme is noncompetively inhibited by inorganic monophosphate (K _i=2.85 mmol/L) and by Cu²⁺, Zn²⁺, Hg²⁺, Sn²⁺, Cd²⁺ ions and strongly by F⁻ ones; it is activated by Ca²⁺, Mg²⁺ and Mn²⁺ ions. The substrate specificity of phytase is broad with the highest affinity to calcium phytate.     

Saccharification of cellulosic substrates byAspergillus niger cellulase

Chemical pre-treatment enhanced saccharification of agricultural lignocellulosic residues usingAspergillus niger cellulase by between 16 and 38%. Maximum saccharification (76%) of alkali-treated bagasse was with 0.5% substrate over 48 h using 0.3 U enzyme/ml.

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Résumé Le pré-traitement chimique augmente de 16 à 38% la saccharification de résidus agricoles ligno-cellulosiques lorsque l'on utilise la cellulase d'Aspergillus niger. On a obtenu la saccharification maximum (76%) de la bagasse traitée à l'alcali avec 0.5% de substrat en 48 h de traitement avec 0.3 unités d'enzyme par ml.

     

Sodium gluconate production byAspergillus niger with intermittent broth replacement

Intermittent broth replacement was carried out to enhance the productivity and purity of sodium gluconate usingAspergillus niger by reducing the concentration of unmetabolized glucose. As inoculum size increased, length of lag phase was shortened and high initial production rate of sodium gluconate was achieved. However, too high inoculum concentration lowered productivity during the later stage of fermentation and increased residual glucose at the end of cultivation. When culture broth was replaced intermittently with distilled water, fresh medium, or recycled medium for comparison with traditional fermentation method, production of sodium gluconate was enhanced more than 1.5 fold and active production period could be prolonged without residual glucose. In addition, productivity was maintained at a level higher than 6 g/L/nr. Therefore, it was found that the reduction of biomass and viscosity by intermittent broth replacement could enhance the productivity.     

Solid state fermentation of dried citrus peel byAspergillus niger

Summary Solid state fermentation of dried citrus peel was studied in a packed bed reactor withAspergillus niger QH-2. When no nitrogen source was added the growth was limited. When urea and/or ammonium sulphate were added in the proportion of 0.63 g N/100 g solid dry substrate a diauxie was observed and the final content of crude protein was higher than 10%. The results of a factorial design show that ammonium sulphate has a significant effect on the specific growth rate.     

Role of glutamine synthetase in citric acid fermentation byAspergillus niger

The activity of glutamine synthetase fromAspergillus niger was significantly lowered under conditions of citric acid fermentation. The intracellular pH of the organism as determined by bromophenol blue dye distribution and fluorescein diacetate uptake methods was relatively constant between 6·0–6·5, when the pH of the external medium was varied between 2·3–7·0.Aspergillus niger glutamine synthetase was rapidly inactivated under acidic pH conditions and Mn²⁺ ions partially protected the enzyme against this inactivation. Mn²⁺-dependent glutamine synthetase activity was higher at acidic pH (6·0) compared to Mg²⁺-supported activity. While the concentration of Mg²⁺ required to optimally activate glutamine synthetase at pH 6·0 was very high (≥ 50 mM), Mn²⁺ was effective at 4 mM. Higher concentrations of Mn²⁺ were inhibitory. The inhibition of both Mn²⁺ and Mg²⁺-dependent reactions by citrate, 2-oxoglutarate and ATP were probably due to their ability to chelate divalent ions rather than as regulatory molecules. This suggestion was supported by the observation that a metal ion chelator, EDTA also produced similar effects. Of the end-products of the pathway, only histidine, carbamyl phosphate, AMP and ADP inhibitedAspergillus niger glutamine synthetase. The inhibitions were more pronounced when Mn²⁺ was the metal ion activator and greater inhibition was observed at lower pH values. These results permit us to postulate that glutamine synthesis may be markedly inhibited when the fungus is grown under conditions suitable for citric acid production and this block may result in delinking carbon and nitrogen metabolism leading to acidogenesis     

Evaluation of alkali treatment for biodegradation of corn cobs byAspergillus niger

Anstract Effect of NaOH pretreatment on the biodegradation of corn cobs for the production of cellulase and protein was studied usingAspergillus niger. Delignification of cobs with NaOH remarkaby increased the production of cellulase and protein. Treatment of cobs with 2% NaOH was found to be the best with respect to their susceptibility to biodegradation for maximum production of cellulose 1,4-β-cellobiosidase, cellulase, β-glucosidase soluble protein and crude protein; this also led to the highest protein recovery, maximum cellulose utilization and also for the maximum degradation of substrate.     

The production of homogentisic acid out of phenylacetic acid byAspergillus niger

Summary With the aid of the replacement culture technique it was found that a strain ofAspergillus niger was able to produce homogentisic acid out of phenylacetic acid with a yield upto 15.7 % of the theoretical amount.Evidence is in favour of the view that homogentisic acid is a normal intermediate in the oxidation of the said substrate.     

Production of protein and cellulase byAspergillus niger in solid state culture

Summary Single-cell protein (SCP) and cellulase production byAspergillus niger AS-101, grown on alkalitreated corn cobs, was studied under various cultural conditions. The maximum yields of SCP and cellulase, under solid-state fermentation were obtained when the culture was incubated at pH 5.5 for 18 d at 30°C with 12% substrate.

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Effect des conditions de culture sur la production de protéines et de cellulases par Aspergillus niger en milieu solide

Résumé La production de protéine d'origine unicellulaire (POU) et de cellulase parAspergillus niger AS-101, cultivé sur des épis de maïs traités à l'alcali, est étudiée sous diverses conditions de culture. Les rendements maximum en POU et en cellulase par fermentation en milieu solide sont obtenus lorsque la culture est incubée à pH 5.5 pendant 18 jours à 30°C et avec une concentration en substrat de 12%.

     

Effect of gamma irradiation pretreatment on biodegradation of forest lignocelluloses byAspergillus niger

Summary Biodegradation of gamma irradiated lignocellulosic forest wastes byAspergillus niger exhibited higher release of reducing sugars and proteins. The pretreated needles ofCedrus deodara showed maximum release of sugars(8%) among fifteen materials studied whereas, only 1.5% sugars were recorded from wood ofPopulus ciliata. In all, this treatment increased the susceptibility of all lignocelluloses to the hydrolytic action of the fungus and resulted in increase in the level of sugars released in comparison of untreated materials. The needles ofCedrus andPinus and woods ofPopulus andRobinia emerged as promising raw materials for fermentation.     

Evaluation of chemical pre-treatment for biodegradation of agricultural lignocellulosic wastes byAspergillus niger

Summary Chemical pre-treatment with NaOH, NaClO₂ or peracetic acid of wheat straw, bagasse, corn cobs and groundnut shells increased cellulose utilization, cellulase activity and protein content ofAspergillus niger AS-101 used as a test biodegradative microorganism.

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Resumen El pre-tratamiento químico con NaOH, NaClO2 o àcido peracético de paja de trigo, bagazo, mazorcas de maíz y cáscaras de cacahuete, incrementó la utilización de celulosa, la actividad celulásica y el contenido en proteínas deAspergillus niger AS-101.

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Résumé Le prétraitement chimique par NaOH, NaClO₂ ou par l'acide peracétique de la paille de froment, de la bagasse, des épis de maïs ou des coquilles d'arachide, augmente l'utilisation de la cellulose, l'activité de la cellulase et le contenu en protéines d'Aspergillus niger AS-101, utilisé comme souche test dans la biodégradation par les microorganismes.

     

Influence of environmental conditions on the production of pectinesterase and polygalacturonase byAspergillus niger

Summary A pectinesterase- and polygalacturonase-producer strain ofAspergillus niger showed a diauxic growth when cultivated in a medium containing pectin as the carbon source. Diauxie was suppressed by adding yeast extract to the medium. The specific production of polygalacturonase was decreased by increasing the initial concentration of pectin, while that of pectinesterase was enhanced with up to 15 g pectin/l. Yeast extract increased the production of biomass but decreased that of the enzymes. The temperature of incubation (25 to 40°C) did not affect the production of biomass or that of polygalacturonase, but production of pectinesterase was markedly affected, 30°C being the optimal temperature.

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Influence de la composition du milieu et les conditions des cultures sur la production de pectinestérase et polygalacturonase par Aspergillus niger

Résumé Aspergillus niger que produit pectinestérase et polygalacturonase, quand est developpé dans un milieu avec pectine comme source de carbone a presenté le phenomene de diauxie. Cela ne succedait pas quand le milieu avait aussi extrait de levure dans sa composition. L'effect de la concentration initiale de pectine sur la production specifique des enzymes des cellules (unité d'enzyme g^–1 de cellules poid sec) a été différent selon la phase du developpement de la culture, sugérant une rélation étroite parmi la synthèse des enzymes et l'état physiologique des cellules. L'augmentation de la concentration de pectine a troublé beaucoup la production spécifique de polygalacturonase. Au contraire, jusqu'à une concentration de 15 g/l a augmenté la synthèse de pectinestérase. La grandeur et structure des pellets ont changé avec la variation de la concentration de pectine, ces modifications peuvent avoir eu influence sur les valeurs des coefficients de transfert de masse et en conséquence sur la cinétique de production des enzymes. L'augmentation de la concentration iniciale d'extrait de levure dans le milieu que contenait pectine comme source de carbone a augmenté la production de biomasse, mais a baisse la synthèse des enzymes surtout celle de pectinestérase. Températures parmi 25°C et 40°C n'ont pas modifié la production de biomasse et non plus polygalacturonase. Au contraire la production de pectinesterase a été bien différente selon la température utilisé ayant un optimum de 30°C.

     

Effect of cadmium and copper on the production of citric acid byAspergillus niger

Copper and cadmium inhibited the growth as well as citric acid production (depending on the heavy metal concentrations) by citric-acid-producingAspergillus niger. Activity of citrate synthase was connected with citrate synthesis in the absence as well as in the presence of heavy metals. The activity of aconitase, and both NAD- and NADP-isocitrate dehydrogenases was strongly inhibited by copper. The contents of DNA and proteins in the cells decreased but the contents of lipids and polysaccharides increased considerably in the presence of both heavy metals.     

The role of acid phosphatase activity during enzymic dephosphorylation of phytates byAspergillus niger phytase

Acid phosphatase present in preparations ofAspergillus niger phytase accelerated dephosphorylation of sodium phytate. Its influence on the reaction rate and distribution ofmyo-inositol phosphates was most apparent at low pH value (2.5) and when acid-hydrolysed substrate was de-esterified enzymatically. With partly purified phytase, the predominant inositol form was tetraphosphate but a preparation having acid phosphatase activity caused an even distribution of lower inositol phosphates after a few hours.     

An indirect method for studying the fine control of citric acid formation byAspergillus niger

Summary During the main period of citric acid fermentation byAspernillus niger product formation is easily determinable from oxygen uptake and carbon dioxide output rates. This was applied in a study on the response of citrate formation to shifts of some environmental parameters of known importance during fermentation.     

Comparison of citric acid production byAspergillus niger immobilized in gels and cryogels of polyacrylamide

Aspergillus niger was immobilized in cryogels and in conventional gels of polyacrylamide. The growth of cells entrapped in two kinds of gels and the production of citric acid by the immobilized cells were investigated and compared. Cells immobilized in cryogels were more suitable for citric acid production.     

Citric acid production byAspergillus niger using media containing low concentrations of glucose or corn starch

By using an appropriate ratio of carbon source to mineral components, we obtained comparable citrate yields in media containing different concentrations of glucose. The enzyme system of inoculum passed on gradually from “growth” state to “production” state during the mould growth. In the starch medium, the critical factors of citric acid production are the aeration efficiency of the medium and the amylase formation of the strain. The air interruption exhibited a prolonged inhibition of the production rate but not of the citrate yield in glucose medium while those parameters in starch medium containing excessive urea were briefly but severely inhibited. After being affected by these unfavorable conditions, the production activity ofAspergillus niger could be restored by applying an appropriate fermentation process.     

Solid-state fermentation of carob pods byAspergillus niger for protein production: effect of particle size

Two strains ofAspergillus niger were cultured in solid-state fermentation system on carob pods ground from 1.25 to 8 mm diam. A particle size of 2.5 mm gave the highest protein content of the final product (20%, w/w) and 52% of the total soluble carbohydrates were utilized. The total tannin concentration of the carob pods decreased by 83% in 4 days of fermentation.T. Smail and O. Salhi are with the Laboratory of Microbiology, U.R.B.A.F., Institute of Biology, Tizi-Ouzou University, Algeria. J.S. Knapp is with the Department of Microbiology, The University of Leeds, Leeds LS2 9JT, UK;