Ultrastructural Localization of Cellulase in Diplodia Maydis and in D. Maydis-Infected Com Stalk Tissue

1977 ◽  
Vol 35 ◽  
pp. 454-455
Author(s):  
Judith A. Murphy ◽  
Mary R. Thompson ◽  
A.J. Pappelis
Keyword(s):  
Culture Medium ◽  
Cupric Oxide ◽  
Cellulase Activity ◽  
Growth Period ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Low Ph ◽  
Cellulolytic Enzymes ◽  
Cellulolytic Activity ◽  
X Ray

BeMiller et.al.(l) found that D. maydis did not have the solubilizing enzyme C1. They reported that D- maydis exhibited cellulolytic activity constitutively, and hypothesized that the cellulolytic enzymes were attached to fungal hyphal surfaces because they found cellulase released to the culture medium only after the growth period, when available cellulose had been used up.The purpose of this study was to determine the location of cellulolytic enzymes (EC 3.2.1.4; beta-1,4-glucan glucanohydrolase) in D. maydis and D. maydis-infected corn tissue at the ultrastructural level.Cellulase activity produces glucose as an end product which will reduce cupric oxide and can be visualized with an EM because it is electron dense and the Cu component can be verified with x-ray analysis(Figs.l,2). After thorough washing, samples fixed in aldehydes are incubated in a substrate mixture at a low pH. The enzyme is activated and reducing sugar is released. The sample is then reacted with Benedict's solution at a high temperature, allowing CuO crystals to be deposited at the site of reaction.

Ultrastructural localization of cellulase in Trichoderma reesei using immunocytochemistry and enzyme cytochemistry.

1983 ◽  
Vol 31 (12) ◽  
pp. 1363-1366 ◽  
Author(s):  
C M Chapman ◽  
J R Loewenberg ◽  
M J Schaller ◽  
J E Piechura
Keyword(s):  
Endoplasmic Reticulum ◽  
Enzyme Activity ◽  
Trichoderma Reesei ◽  
Growth Medium ◽  
Culture Medium ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Enzyme Cytochemistry ◽  
Specific Staining ◽  
Cellulase Complex

Two components of the cellulase complex (E.C. 3.2.1.4) of the fungus Trichoderma reesei were localized at the ultrastructural level. Immunocytochemistry and enzyme cytochemistry demonstrated that cellobiohydrolase and beta-1,4 glucanase were localized within cisternae of endoplasmic reticulum and within membrane complexes of cellulose-grown hyphae. Both enzymes were also present in the culture medium. Glucose-grown control hyphae lacked enzyme-specific staining, and no enzyme activity was detected in the growth medium.

scholarly journals Isolation, Cellulase Activity Test and Molecular Identification of Selected Cellulolytic Bacteria Indigenous Rice Bran

2018 ◽  
Vol 18 (3) ◽  
pp. 514 ◽  
Author(s):  
Akyunul Jannah ◽  
Aulanni`am Aulanni`am ◽  
Tri Ardyati ◽  
Suharjono Suharjono
Keyword(s):  
Rice Bran ◽  
Waste Product ◽  
Cellulase Activity ◽  
Raw Material ◽  
Cellulolytic Enzymes ◽  
Cellulolytic Activity ◽  
Cellulolytic Bacteria ◽  
Endoglucanase Activity ◽  
Activity Test ◽  
Rice Milling

Rice bran is the waste product of rice milling which is abundant in Indonesia, it can be used as a raw material for the manufacture of bioethanol by fermentation. Before being fermented, rice bran must be hydrolyzed into glucose by biomass degrading. This study was aimed to isolate indigenous cellulolytic bacteria from rice bran as producer of cellulolytic enzymes and resulted in 22 bacterial isolates that demonstrated cellulolytic activity being identified. Among them, BE 8 and BE 14 isolates showed the highest endoglucanase activity at 2.16 and 1.31 U/mL respectively. Identification of the 16S rDNA showed that BE 8 belongs to Bacillus subtilis and BE 14 in Bacillus cereus.

scholarly journals Immobilization of (Aqueous) Cations in Low pH M-S-H Cement

2021 ◽  
Vol 11 (7) ◽  
pp. 2968
Author(s):  
Maximilian R. Marsiske ◽  
Christian Debus ◽  
Fulvio Di Lorenzo ◽  
Ellina Bernard ◽  
Sergey V. Churakov ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Metal Cations ◽  
Growth Period ◽  
Magnesium Silicate ◽  
Low Ph ◽  
Secondary Phases ◽  
X Ray ◽  
Early Stages ◽  
Total Metal ◽  
Wide Range

Incorporation of heavy metal ions in cement hydrates is of great interest for the storage and immobilization of toxic, hazardous, and radioactive wastes using cementitious matrix. Magnesium silicate hydrate (M-S-H) is a low pH alternative cementitious binder to commonly used Portland cement. Low pH cements have been considered as promising matrix for municipal and nuclear waste immobilization in the last decades. It is however crucial to assure that the incorporation of secondary ions is not detrimental for the formation of the hydration products. Herein, we investigate the early stages of formation of M-S-H from electrolyte solutions in presence of a wide range of metal cations (LiI, BaII, CsI, CrIII, FeIII, CoII, NiII, CuI, ZnII, PbII, AlIII). The final solid products obtained after 24 h have been characterized via powder X-ray diffraction (PXRD), attenuated total reflectance-Fourier transformed infrared spectroscopy (FTIR-ATR), elemental analysis via energy-dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), and high-resolution transmission electron microscopy (HR-TEM). In all the experiments, the main precipitated phase after 24 h was confirmed to be M-S-H with a ratio (total metal/Si) close to one. The obtained M-S-H products showed strong immobilization capacity for the secondary metal cations and can incorporate up to 30% of the total metal content at the early stages of M-S-H formation without significantly delaying the nucleation of the M-S-H. It has been observed that presence of Cr, Co, and Fe in the solution is prolonging the growth period of M-S-H. This is related to a higher average secondary metal/total metal ratio in the precipitated material. Secondary phases that co-precipitate in some of the experiments (Fe, Pb, Ni, and Zn) were also effectively trapped within in the M-S-H matrix. Barium was the only element in which the formation of a secondary carbonate phase isolated from the M-S-H precipitates was detected.

X-Ray Microanalysis of Nickel and Cobalt Intensified Peroxidase- Antiperoxidase For Verification of Reaction Sites

1985 ◽  
Vol 43 ◽  
pp. 468-469
Author(s):  
A. Angel ◽  
K. Miller ◽  
V. Seybold ◽  
R. Kriebel
Keyword(s):  
Reaction Mechanisms ◽  
Visual Discrimination ◽  
Osmium Tetroxide ◽  
Ultrastructural Level ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
X Ray ◽  
Insoluble Polymer ◽  
Cytochemical Reaction

Localization of specific substances at the ultrastructural level is dependent on the introduction of chemicals which will complex and impart an electron density at specific reaction sites. Peroxidase-antiperoxidase(PAP) methods have been successfully applied at the electron microscopic level. The PAP complex is localized by addition of its substrate, hydrogen peroxide and an electron donor, usually diaminobenzidine(DAB). On oxidation, DAB forms an insoluble polymer which is able to chelate with osmium tetroxide becoming electron dense. Since verification of reactivity is visual, discrimination of reaction product from osmiophillic structures may be difficult. Recently, x-ray microanalysis has been applied to examine cytochemical reaction precipitates, their distribution in tissues, and to study cytochemical reaction mechanisms. For example, immunoreactive sites labelled with gold have been ascertained by means of x-ray microanalysis.

Ultrastructural Localization of Antigens by Capillary Action Immunocytochemistry

1996 ◽  
Vol 54 ◽  
pp. 40-41
Author(s):  
László G. Kömüves
Keyword(s):  
San Francisco ◽  
Colloidal Gold ◽  
Specific Binding ◽  
Ultrastructural Localization ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Adhesive Tape ◽  
Distilled Water ◽  
Capillary Action ◽  
Rabbit Igg

Light microscopic immunohistochemistry based on the principle of capillary action staining is a widely used method to localize antigens. Capillary action immunostaining, however, has not been tested or applied to detect antigens at the ultrastructural level. The aim of this work was to establish a capillary action staining method for localization of intracellular antigens, using colloidal gold probes.Post-embedding capillary action immunocytochemistry was used to detect maternal IgG in the small intestine of newborn suckling piglets. Pieces of the jejunum of newborn piglets suckled for 12 h were fixed and embedded into LR White resin. Sections on nickel grids were secured on a capillary action glass slide (100 μm wide capillary gap, Bio-Tek Solutions, Santa Barbara CA, distributed by CMS, Houston, TX) by double sided adhesive tape. Immunolabeling was performed by applying reagents over the grids using capillary action and removing reagents by blotting on filter paper. Reagents for capillary action staining were from Biomeda (Foster City, CA). The following steps were performed: 1) wet the surface of the sections with automation buffer twice, 5 min each; 2) block non-specific binding sites with tissue conditioner, 10 min; 3) apply first antibody (affinity-purified rabbit anti-porcine IgG, Sigma Chem. Co., St. Louis, MO), diluted in probe diluent, 1 hour; 4) wash with automation buffer three times, 5 min each; 5) apply gold probe (goat anti-rabbit IgG conjugated to 10 nm colloidal gold, Zymed Laboratories, South San Francisco, CA) diluted in probe diluent, 30 min; 6) wash with automation buffer three times, 5 min each; 7) post-fix with 5% glutaraldehyde in PBS for 10 min; 8) wash with PBS twice, 5 min each; 9) contrast with 1% OSO4 in PBS for 15 min; 10) wash with PBS followed by distilled water for5 min each; 11) stain with 2% uranyl acetate for 10 min; 12) stain with lead citrate for 2 min; 13) wash with distilled water three times, 1 min each. The glass slides were separated, and the grids were air-dried, then removed from the adhesive tape. The following controls were used to ensure the specificity of labeling: i) omission of the first antibody; ii) normal rabbit IgG in lieu of first antibody; iii) rabbit anti-porcine IgG absorbed with porcine IgG.

A Facile Synthesis of Cu2O and CuO Nanoparticles Via Sonochemical Assisted Method

2018 ◽  
Vol 15 (2) ◽  
pp. 209-213 ◽  
Author(s):  
Sathish Mohan Botsa ◽  
Ramadevi Dharmasoth ◽  
Keloth Basavaiah
Keyword(s):  
Diffuse Reflectance Spectroscopy ◽  
Cupric Oxide ◽  
Copper Acetate ◽  
Chemical Properties ◽  
Copper Oxide Nanoparticles ◽  
Reducing Agents ◽  
Sonochemical Method ◽  
Phase Purity ◽  
X Ray ◽  
Cuo Nps

Background: During past two decades, functional nanomaterials have received great attention for many technological applications such as catalysis, energy, environment, medical and sensor due to their unique properties at nanoscale. However, copper oxide nanoparticles (NPs) such as CuO and Cu2O have most widely investigated for many potential applications due to their wide bandgap, high TC, high optical absorption and non-toxic in nature. The physical and chemical properties of CuO and Cu2O NPs are critically depending on their size, morphology and phase purity. Therefore, lots of efforts have been done to prepare phase CuO and Cu2O NPs with different morphology and size. Method: The synthesis of cupric oxide (CuO) and cuprous oxide (Cu2O) NPs using copper acetate as a precursor by varying the reducing agents such as hydrazine sulphate and hydrazine hydrate via sonochemical method. The phase, morphology and crystalline structure of a prepared CuO and Cu2O NPs were investigated by X-ray diffraction (XRD), Fourier transform infrared (FTIR), Field emission scanning electron microscopy (FESEM), Energy dispersive X-ray (EDS) and UV-Visible Diffuse reflectance spectroscopy (DRS). Results: The phase of NPs was tuned as a function of reducing agents.XRD patterns confirmed the formation of pure phase crystalline CuO and Cu2O NPs. FTIR peak at 621 cm-1 confirmed Cu(I)-O vibrations, while CuO vibrations confirmed by the presence of two peaks at 536 and 586 cm-1. Further investigation was done by Raman, which clearly indicates the presence of peaks at 290, 336, 302 cm-1 and 173, 241 cm-1 for CuO and Cu2O NPs, respectively. The FESEM images revealed rod-like morphology of the CuO NPs while octahedral like shape for Cu2O NPs. The presence of elemental Cu and O in stoichiometric ratios in EDS spectra confirms the formation of both CuO and Cu2O NPs. In summary, CuO and Cu2O NPs were successfully synthesized by a sonochemical method using copper acetate as a precursor at different reducing agents. The bandgap of CuO and Cu2O NPs was 2.38 and 1.82, respectively. Furthermore, the phase purity critically depends on reducing agents.

Fluoride-containing bioactive glasses and Bioglass® 45S5 form apatite in low pH cell culture medium

2014 ◽  
Vol 119 ◽  
pp. 96-99 ◽  
Author(s):  
Furqan A. Shah ◽  
Delia S. Brauer ◽  
Nikita Desai ◽  
Robert G. Hill ◽  
Karin A. Hing
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Low Ph ◽  
Cell Culture Medium ◽  
Bioactive Glasses

scholarly journals An Efficient and Improved Methodology for the Screening of Industrially Valuable Xylano-Pectino-Cellulolytic Microbes

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Avtar Singh ◽  
Amanjot Kaur ◽  
Anita Dua ◽  
Ritu Mahajan
Keyword(s):  
Low Cost ◽  
Cost Effective ◽  
Industrial Sector ◽  
Agricultural Residues ◽  
High Yield ◽  
Cellulolytic Enzymes ◽  
Cellulolytic Activity ◽  
Nutrient Media ◽  
First Report ◽  
Primary Screening

Xylano-pectino-cellulolytic enzymes are valuable enzymes of the industrial sector. In our earlier study, we have reported a novel and cost effective methodology for the qualitative screening of cellulase-free xylano-pectinolytic microorganisms by replacing the commercial, highly expensive substrates with agricultural residues, but the microorganisms with xylanolytic, pectinolytic, cellulolytic, xylano-pectinolytic, xylano-cellulolytic, pectino-cellulolytic, and xylano-pectino-cellulolytic potential were obtained. The probability of getting the desired combination was low, so efforts were made to further improve this cost effective methodology for obtaining the high yield of the microbes capable of producing desired combination of enzymes. By inclusion of multiple enrichment steps in sequence, using only practically low cost substrates and without any nutrient media till primary screening stage, this improved novel protocol for screening gave only the desired microorganisms with xylano-pectino-cellulolytic activity. Using this rapid, efficient, cost effective, and improved methodology, microbes with required combination of enzymes can be obtained and the probability of getting the desired microorganisms is cent percent. This is the first report presenting the methodology for the isolation of xylano-pectino-cellulolytic positive microorganisms at low cost and consuming less time.

scholarly journals Controllability of cupric particle synthesis by linear alcohol chain number as additive and pH control in cupric acetate solution using X-ray radiolysis

2019 ◽  
Vol 26 (6) ◽  
pp. 1986-1995 ◽  
Author(s):  
Akinobu Yamaguchi ◽  
Ikuo Okada ◽  
Ikuya Sakurai ◽  
Hirokazu Izumi ◽  
Mari Ishihara ◽  
...  
Keyword(s):  
Aqueous Solutions ◽  
Chain Length ◽  
Cupric Oxide ◽  
Liquid Solution ◽  
Ph Control ◽  
Acetate Solution ◽  
Reaction Route ◽  
X Ray ◽  
Rate Of Reaction ◽  
Oxide Particles

Synthesis and immobilization of caltrop cupric particles onto a Si substrate using X-ray radiolysis directly from a liquid solution of Cu(COOCH3)2 is demonstrated. Caltrop cupric oxide particles are formed in the X-ray radiolysis of aqueous solutions of Cu(COOCH3)2, which also contain methanol, ethanol, 2-propanol or 1-propanol as ^\bulletOH scavenger. The blade lengths of the caltrop particles are dependent on the alcohol chain length. In particular, it was found that an alkyl alcohol whose chain length is longer than four is unable to synthesize any particles in aqueous solutions of Cu(COOCH3)2 in X-ray radiolysis. These results are attributed to the alkyl alcohol chain length influencing the rate of reaction of radicals and determines the solvable ratio of its alcohol into water. In addition, it was found that the synthesized particle geometric structure and composition can also be controlled by the pH of the aqueous solution in the X-ray radiolysis. This study may open a door to understanding and investigating a novel photochemical reaction route induced under X-ray irradiation. The development of the X-ray radiolysis process enables us to achieve the rapid and easy process of synthesis and immobilization of higher-order nano/microstructure consisting of various materials.

scholarly journals First Report of Black Soft Rot of Indian Gooseberry Caused by Syncephalastrum racemosum

Plant Disease ◽  
2004 ◽  
Vol 88 (5) ◽  
pp. 575-575 ◽  
Author(s):  
Neelima Garg ◽  
Om Prakash ◽  
B. K. Pandey ◽  
B. P. Singh ◽  
G. Pandey
Keyword(s):  
Cellulase Activity ◽  
Basal Medium ◽  
Petri Dish ◽  
Soft Rot ◽  
Cellulolytic Activity ◽  
Academic Press ◽  
First Report ◽  
Syncephalastrum Racemosum ◽  
Indian Gooseberry ◽  
Pathogenicity Tests

Indian gooseberry (Emblica officinalis Gaertn.) is a medicinal plant with high nutraceutical value. During November and December 2003, soft rot was noticed on harvested and stored (20 ± 5°C and 65 ± 5% relative humidity) fruits at the experimental farm in Rehmanhera, Lucknow, India (26°50′N, 80°54′E). These fruits had numerous, minute brown necrotic lesions showing white mycelial growth. A pronounced halo of water-soaked, faded tissue surrounded the lesion between the fringe of mycelium and healthy tissues. The rotted surface was covered with a black, powdery layer of spores. On Czapek yeast extract agar, fungal colonies were blackish grey, moderately dense, and covered the entire petri dish. The fungus produced aseptate mycelium. The sporangial heads were 30 to 50 μm in diameter with sporangiospores found linearly within cylindrical sacs (merosporangia) borne on spicules around the columella. Sporangiospores, spherical to cylindrical in shape and borne in chains, measured 3.0 to 5.0 μm long. The fungus was morphologically and physiologically identified as Syncephalastrum racemosum Schr. (2). For pathogenicity tests, healthy fruits (10 replicates) were surface sterilized and punctured inoculated aseptically with 1.0 × 106 conidia and incubated at 20 ± 5°C Typical symptoms of the disease appeared after 4 days. The fungus exhibited a strong level of cellulolytic activity as indicated by prolific growth on Indian gooseberry fiber waste under solid-state fermentation conditions. The level of cellulase activity (1) was 21 filter paper activity unit per ml at 72 hr in culture supernatant of basal medium having carboxymethyl cellulose as the carbon source. The fungus showed resistance to tannins (as much as 2%), since it could grow well in liquid growth medium (Czapek Dox broth) with 2% tannins and aonla juice with 1.8% tannins. Since Indian gooseberry is rich in fiber (2.5 to 3.4%) and tannins (1.5 to 2.0%), this may be an important pathogen. To our knowledge, this is the first report of the occurrence of Syncephalastrum racemosum on Indian gooseberry fruits. References: (1) T. K. Ghose. Pure Appl. Chem. 59(2):257, 1987. (2) J. I. Pitt and A. D. Hocking. Fungi and Food Spoilage. Academic Press. North Ryde, Australia, 1985.

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