Influence of pH on endo- and exo-pectinase production by Aspergillus sp. CH-Y-1043

1991 ◽  
Vol 37 (12) ◽  
pp. 912-917 ◽  
Author(s):  
Guillermo Aguilar ◽  
Blanca A. Trejo ◽  
Juan M. García ◽  
Carlos Huitrón
Keyword(s):  
Fungal Growth ◽  
Protein Band ◽  
Ph Values ◽  
Sds Page ◽  
Pectin Esterase ◽  
Aspergillus Sp ◽  
Pectinase Production ◽  
Key Words Aspergillus ◽  
Maximal Production

Aspergillus sp. CH-Y-1043 grown on pectins with various degrees of esterification produces endo- and exo-pectinases at pH values as low as 2.5. Maximal production was attained at this pH, although fungal growth only approximated 50% of that obtained at higher pH values. Endopectinase was produced at pH 2.5–3.5 when the fungus was grown on low degree esterified pectin. With higher degree esterified pectin this enzyme was produced at all pH values analyzed. Exopectinase production was less affected by pH values. Still, maximal production was also reached at pH 2.5–3.5. Exopectinase was found to be associated to the cell and could be released after incubation at different pH values, whereas endo pectinase was not detected in the cellular fraction. Results confirmed by SDS–PAGE coupled with in situ activity assays in pectin–agarose gels allowed the identification of a protein band corresponding to endopectinase and a band with pectin esterase activity. Stability of Aspergillus sp. CH-Y-1043 pectinases at various pH values was also evaluated. Key words: Aspergillus sp. CH-Y-1043, extreme acidic pH pectinase production, in situ pectinase detection, cell-associated exopectinase.

scholarly journals A murine hybridoma with large cytoplasmic inclusions of kappa light chains.

1987 ◽  
Vol 165 (2) ◽  
pp. 578-583 ◽  
Author(s):  
D C Reason
Keyword(s):  
Cell Line ◽  
Inclusion Bodies ◽  
Protein Band ◽  
Light Chains ◽  
Cytoplasmic Inclusions ◽  
Single Protein Band ◽  
Sds Page ◽  
Single Protein ◽  
Kappa Light Chains

A murine hybridoma cell line has been established that consistently forms large cytoplasmic inclusions. These structures bind antibody specific for mouse kappa L chain when stained in situ. SDS-PAGE analysis of isolated inclusion bodies produce a single protein band of approximately 26,000 Mr that reacts with anti-kappa antibody when transferred to nitrocellulose. No carbohydrate was detected in association with the purified protein. These data are consistent with the intracellular retention and deposition of complete kappa L chain protein.

scholarly journals Isolation and partial characterization of a lectin from Bauhinia pentandra (bong) vog. Ex. Steua.

2001 ◽  
Vol 13 (3) ◽  
pp. 262-269 ◽  
Author(s):  
ANDRÉ LUIS COELHO DA SILVA ◽  
ANA CECÍLIA GÓES HORTA ◽  
RENATO DE AZEVEDO MOREIRA
Keyword(s):  
Protein Band ◽  
Carbohydrate Specificity ◽  
Original Activity ◽  
Divalent Metal Cations ◽  
Ph Values ◽  
Seed Flour ◽  
Sds Page ◽  
Low Levels ◽  
Sepharose 4B

Bauhinia pentandra (Bong) Vog. ex. Steua seeds were investigated with respect to phenologic aspects (size, mass, hilum and length) and with respect to their chemical composition. The total nitrogen content of the seed flour was determined, and the flour was extracted in different pH values. A lectin was isolated from the seeds by Sepharose-4B affinity chromatography. The homogeneity of the lectin was demonstrated by SDS-PAGE in the presence of beta-mercaptoethanol. Only one protein band with an apparent molecular mass of 30 kDa was found. The B. pentandra lectin showed a carbohydrate specificity for D-galactose, a requirement for divalent metal cations (Ca2+ and Mn2+) for full activity and amino acid composition with a high content of aspartic acid, glutamic acid and alanine and low levels of methionine, cysteine and tryptophan. The lectin agglutinated rabbit and human A group erythrocytes and was relatively stable to heat treatment, retaining half of its original activity after 60 min at 70 ºC.

scholarly journals Effect of Ocular Hypertension on D-β-Aspartic Acid-Containing Proteins in the Retinas of Rats

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Takashi Kanamoto ◽  
Takashi Tachibana ◽  
Yasushi Kitaoka ◽  
Toshio Hisatomi ◽  
Yasuhiro Ikeda ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Ocular Hypertension ◽  
Aspartic Acid ◽  
Atp Synthase ◽  
Wistar Rats ◽  
Protein Band ◽  
Mass Spectrometry Analysis ◽  
Liquid Chromatography Mass Spectrometry ◽  
Spectrometry Analysis ◽  
Sds Page

Purpose. To investigate the effect of ocular hypertension-induced isomerization of aspartic acid in retinal proteins. Methods. Adult Wistar rats with ocular hypertension were used as an experimental model. D-β-aspartic acid-containing proteins were isolated by SDS-PAGE and western blot with an anti-D-β-aspartic acid antibody and identified by liquid chromatography-mass spectrometry analysis. The concentration of ATP was measured by ELISA. Results. D-β-aspartic acid was expressed in a protein band at around 44.5 kDa at much higher quantities in the retinas of rats with ocular hypertension than in those of normotensive rats. The 44.5 kDa protein band was mainly composed of α-enolase, S-arrestin, and ATP synthase subunits α and β, in both the ocular hypertensive and normotensive retinas. Moreover, increasing intraocular pressure was correlated with increasing ATP concentrations in the retinas of rats. Conclusion. Ocular hypertension affected the expression of proteins containing D-β-aspartic acid, including ATP synthase subunits, and up-regulation of ATP in the retinas of rats.

scholarly journals Bioactive Metabolites from the Endophytic Fungus Aspergillus sp. SPH2

2021 ◽  
Vol 7 (2) ◽  
pp. 109
Author(s):  
Viridiana Morales-Sánchez ◽  
Carmen E. Díaz ◽  
Elena Trujillo ◽  
Sonia A. Olmeda ◽  
Felix Valcarcel ◽  
...  
Keyword(s):  
Endophytic Fungus ◽  
Plant Pathogens ◽  
Insect Pests ◽  
Ethyl Acetate Extract ◽  
Rhopalosiphum Padi ◽  
Fungal Growth ◽  
Bioactive Metabolites ◽  
Endemic Plant ◽  
Antifungal Effects ◽  
Aspergillus Sp

In the current study, an ethyl acetate extract from the endophytic fungus Aspergillus sp. SPH2 isolated from the stem parts of the endemic plant Bethencourtia palmensis was screened for its biocontrol properties against plant pathogens (Fusarium moniliforme, Alternaria alternata, and Botrytis cinerea), insect pests (Spodoptera littoralis, Myzus persicae, Rhopalosiphum padi), plant parasites (Meloidogyne javanica), and ticks (Hyalomma lusitanicum). SPH2 gave extracts with strong fungicidal and ixodicidal effects at different fermentation times. The bioguided isolation of these extracts gave compounds 1–3. Mellein (1) showed strong ixodicidal effects and was also fungicidal. This is the first report on the ixodicidal effects of 1. Neoaspergillic acid (2) showed potent antifungal effects. Compound 2 appeared during the exponential phase of the fungal growth while neohydroxyaspergillic acid (3) appeared during the stationary phase, suggesting that 2 is the biosynthetic precursor of 3. The mycotoxin ochratoxin A was not detected under the fermentation conditions used in this work. Therefore, SPH2 could be a potential biotechnological tool for the production of ixodicidal extracts rich in mellein.

scholarly journals Competition between Metals for Binding to Methanobactin Enables Expression of Soluble Methane Monooxygenase in the Presence of Copper

2014 ◽  
Vol 81 (3) ◽  
pp. 1024-1031 ◽  
Author(s):  
Bhagyalakshmi Kalidass ◽  
Muhammad Farhan Ul-Haque ◽  
Bipin S. Baral ◽  
Alan A. DiSpirito ◽  
Jeremy D. Semrau
Keyword(s):  
Methane Monooxygenase ◽  
High Specificity ◽  
Copper Uptake ◽  
Content Type ◽  
Soluble Methane Monooxygenase ◽  
Sds Page ◽  
Genes Encoding ◽  
Key Factor ◽  
Membrane Bound

ABSTRACTIt is well known that copper is a key factor regulating expression of the two forms of methane monooxygenase found in proteobacterial methanotrophs. Of these forms, the cytoplasmic, or soluble, methane monooxygenase (sMMO) is expressed only at low copper concentrations. The membrane-bound, or particulate, methane monooxygenase (pMMO) is constitutively expressed with respect to copper, and such expression increases with increasing copper. Recent findings have shown that copper uptake is mediated by a modified polypeptide, or chalkophore, termed methanobactin. Although methanobactin has high specificity for copper, it can bind other metals, e.g., gold. Here we show that inMethylosinus trichosporiumOB3b, sMMO is expressed and active in the presence of copper if gold is also simultaneously present. Such expression appears to be due to gold binding to methanobactin produced byM. trichosporiumOB3b, thereby limiting copper uptake. Such expression and activity, however, was significantly reduced if methanobactin preloaded with copper was also added. Further, quantitative reverse transcriptase PCR (RT-qPCR) of transcripts of genes encoding polypeptides of both forms of MMO and SDS-PAGE results indicate that both sMMO and pMMO can be expressed when copper and gold are present, as gold effectively competes with copper for binding to methanobactin. Such findings suggest that under certain geochemical conditions, both forms of MMO may be expressed and activein situ. Finally, these findings also suggest strategies whereby field sites can be manipulated to enhance sMMO expression, i.e., through the addition of a metal that can compete with copper for binding to methanobactin.

PROTEIN PROFILING OF DIFFERENT PLANT TISSUES FROM HERB PHYLLANTHUS NIRURI

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Ainul Mardhiah Mohd Nail ◽  
Noor Hasniza Md Zin
Keyword(s):  
Molecular Weight ◽  
Protein Band ◽  
Protein Profiling ◽  
Protein Extract ◽  
Phyllanthus Niruri ◽  
Two Dimensional Gel Electrophoresis ◽  
Plant Parts ◽  
Sds Page ◽  
Isoelectric Points

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties.  In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa.

scholarly journals ATP citrate-lyase and glycogen synthase kinase-3β in 3T3-L1 cells during differentiation into adipocytes

1994 ◽  
Vol 300 (2) ◽  
pp. 477-482 ◽  
Author(s):  
W B Benjamin ◽  
S N Pentyala ◽  
J R Woodgett ◽  
Y Hod ◽  
D Marshak
Keyword(s):  
Glycogen Synthase ◽  
Protein Band ◽  
Glycogen Synthase Kinase ◽  
Fat Cells ◽  
Apparent Mass ◽  
Atp Citrate Lyase ◽  
Protein Levels ◽  
Citrate Lyase ◽  
Sds Page ◽  
Dependent Protein

ATP citrate-lyase (CL), acetyl-CoA carboxylase (ACC) and glycogen synthase kinase-3 beta (GSK-3 beta) levels were measured in cytosol from 3T3-L1 cells during differentiation from fibroblasts into fat-cells. Protein levels were estimated from immunoblots using specific antisera. Cytosol from confluent cells contain significant amounts of GSK-3 beta, which fell during differentiation of these cells into adipocytes. CL from confluent cells was found to be mostly in the form of a single protein band of apparent mass 110 kDa. Levels of CL and ACC increased during cell differentiation into adipocytes. During the first 3 days of differentiation, CL migration changed, and it was expressed as a complex of protein bands of apparent mass 110 kDa, 113 kDa and 115 kDa. At later stages of differentiation, when these cells had assumed the phenotype of fat-cells, they expressed CL mainly as protein bands of 110 and 113 kDa. When samples containing these bands were treated with alkaline phosphatase, the 113 kDa protein band collapsed into the 110 kDa species. This suggests that the slower-migrating species of CL is a higher-order phosphorylation state of the same protein. Furthermore, when purified CL, mostly expressed as the 110 kDa species, was phosphorylated with cyclic AMP-dependent protein kinase alone or together with GSK-3 and resolved by SDS/PAGE, the phosphorylated CL now migrated more slowly as the 113 kDa and 115 kDa forms. CL phosphorylation was hormone-regulated, since, in samples from fat-cells that had the complex two-band pattern, when cultured in medium without serum or hormones, CL migration reverted to a single band of 110 kDa, similar to confluent cells. Treatment of these ‘down-regulated’ cells with insulin rapidly induced substantial amounts of the 113 kDa species, with a concomitant decrease in the 110 kDa species.

scholarly journals The venom and the toxicity of Pelagia noctiluca (Cnidaria: Scyphozoa). A review of three decades of research in Italian laboratories and future perspectives

2015 ◽  
Vol 88 (2) ◽  
Author(s):  
Rossana Morabito ◽  
Angela Marino ◽  
Giuseppina La Spada ◽  
Luigi Pane ◽  
Gian Luigi Mariottini
Keyword(s):  
Neuroblastoma Cells ◽  
Reactive Oxygen Species Generation ◽  
Cell Membrane Permeability ◽  
Crude Venom ◽  
Ph Values ◽  
Pelagia Noctiluca ◽  
Future Challenges ◽  
Induced Apoptosis ◽  
Discharging Capacity

Recurrent outbreaks of <em>Pelagia noctiluca</em> and health problems consequent to stings were recorded during the last decades. This phenomenon forced some Italian University laboratories to study this cnidarian. The first studies concerned the distribution, biochemical composition and morphology of nematocysts of <em>Pelagia noctiluca</em>. The discharge mechanism of nematocysts was defined starting from early 1980s when enzymes, cations, anions, and pH were observed to have an influence on this process. Notably, trypsin, extreme pH values, some anions (I<sup>–</sup>, Cl<sup>–</sup>, SCN<sup>–</sup>), and thioglycolate were seen to induce, while La<sup>3+</sup> and Gd<sup>3+</sup> to prevent, nematocyst discharge. The discharge of both <em>in situ</em> and isolated nematocyst was found to be Ca<sup>2+</sup> dependent. Furthermore, <em>Pelagia noctiluca</em> nematocysts were seen to retain their discharging capacity in distilled water. The toxicological evaluations were carried out mainly using the crude venom from <em>Pelagia noctiluca</em> because, unfortunately, to date the composition of venom remains unknown. Hemolytic and cytotoxic properties of crude venom have been evaluated on erythrocytes and cultured guinea-pig fibroblasts, mouse fibroblasts, and cancer (neuroblastoma) cells. The activity of <em>Pelagia noctiluca</em> venom on other cnidarians has been also assessed. The crude venom induced apoptosis by reactive oxygen species generation and decrease in mitochondrial transmembrane potential, loss of mitochondrial integrity, and alteration of cell membrane permeability. A pore-forming action mechanism on mitochondrial membrane with oxidative damage was also suggested. The protective activity of some compounds against envenomations has been also evaluated. Future challenges will concern the attempts to characterize the venom and to perform a wider screening of cytotoxicity induced to normal and cancer cells.

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