Effect of metabolites on ε-N-hydroxylysine formation in cell-free extracts of Aerobacter aerogenes 62-1

1977 ◽  
Vol 55 (6) ◽  
pp. 625-629 ◽  
Author(s):  
G. J. Murray ◽  
G. E. D. Clark ◽  
M. A. Parniak ◽  
T. Viswanatha
Keyword(s):  
Enzymatic Activity ◽  
Hydroxy Derivative ◽  
Differential Centrifugation ◽  
Hydroxylase Activity ◽  
Aerobacter Aerogenes ◽  
Lactate And Pyruvate ◽  
First Time

The conversion of L-lysine to its corresponding ε-N-hydroxy derivative has been achieved for the first time by cell-free extracts of Aerobacter aerogenes 62-1. Partial fractionation by differential centrifugation (at 12 000 × g) revealed that both supernatant and pellet are essential for maximum enzymatic activity. The ω-N-hydroxylase (EC 1.14.99) was found to function optimally at pH 7–7.5 and exhibited an apparent Km of about 75 μM for L-lysine. L(+)-Lactate or DL-lactate and pyruvate greatly stimulate the ω-N-hydroxylase activity. The system is strongly inhibited by arsenite and sulfite.

Cytotoxic Effects of Ferula Latisecta on Human Glioma U87 Cells

Drug Research ◽  
2019 ◽  
Vol 69 (12) ◽  
pp. 665-670 ◽  
Author(s):  
Mohammad Jalili-Nik ◽  
Hamed Sabri ◽  
Ehsan Zamiri ◽  
Mohammad Soukhtanloo ◽  
Mostafa Karimi Roshan ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Gelatin Zymography ◽  
Annexin V ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Ic50 Values ◽  
Induced Apoptosis ◽  
Natural Herb ◽  
First Time ◽  
U87 Cells

AbstractGlioblastoma multiforme (GBM) is the fatal type of astrocytic tumors with a survival rate of 12 months. The present study, for the first time, evaluated the cytotoxic impacts of Ferula latisecta (F. latisecta) hydroalcoholic extract on U87 GBM cell line. The MTT assay measured the cellular toxicity following 24- and 48 h treatment with various doses of F. latisecta (0–800 μg/mL). Apoptosis was evaluated by an Annexin V/propidium iodide (PI) staining 24 h after treatment by F. latisecta. Moreover, to determine the cellular metastasis of U87 cells, we used a gelatin zymography assay (matrix metalloproteinase [MMP]-2/-9 enzymatic activity). The outcomes showed that F. latisecta mitigated the viability of U87 cells in a concentration- and time-dependent manner with IC50 values of 145.3 and 192.3 μg/mL obtained for 24- and 48 h treatments, respectively. F. latisecta induced apoptosis in a concentration-dependent manner after 24 h. Also, MMP-9 activity was significantly decreased following 24 h after treatment concentration-dependently with no change in MMP-2 enzymatic activity. This study showed that F. latisecta induced cytotoxicity and apoptosis, and mitigated metastasis of U87 GBM cells. Hence, F. latisecta could be beneficial as a promising natural herb against GBM after further studies.

scholarly journals Intracellular Distribution of 5' Nucleotidase in Rat Liver

1958 ◽  
Vol 4 (4) ◽  
pp. 373-376 ◽  
Author(s):  
Gaston de Lamirande ◽  
Claude Allard ◽  
Antonio Cantero
Keyword(s):  
Enzymatic Activity ◽  
Rat Liver ◽  
Biochemical Analysis ◽  
Inorganic Phosphorus ◽  
Intracellular Distribution ◽  
Differential Centrifugation ◽  
Nucleotidase Activity ◽  
Rat Liver Cells ◽  
Cell Fractions

The intracellular distribution of 5' nucleotidase was investigated in rat liver by biochemical analysis of cell fractions obtained by differential centrifugation. The enzymatic activity was measured by determination of the inorganic phosphorus liberated from 5' nucleotides. The 5' nucleotidase activity was mainly found in the nuclear and microsomal fractions. An attempt to extract the enzyme from these fractions with Mg++ ion solutions was unsuccessful. It is concluded that 5' nucleotidase is actually present in the nuclear and microsomal fractions of rat liver cells.

Nucleic acid and protein content of Bact . lactis aerogenes . III. Effect of certain drugs

1962 ◽  
Vol 155 (961) ◽  
pp. 599-603
Keyword(s):  
Nucleic Acid ◽  
Protein Content ◽  
Cell Size ◽  
Free Medium ◽  
Aerobacter Aerogenes ◽  
First Time ◽  
Drug Free

The effect of chloramphenicol, 5·aminoacridine or streptomycin on cell size and on the RNA, DNA and protein content of Bact . lactis aerogenes ( Aerobacter aerogenes ) has been investigated during the course of the first subculture in drag. The behaviour when fully adapted strains are returned to drug-free medium for the first time has also been examined. Results are discussed in terms of the effect of the abnormal conditions on the cell.

Enzymatic Activity and Expression of Cytochrome P450 LAω within Intrasplenically Transplanted Fetal Hepatocytes in Spontaneously Hypertensive Rats

1997 ◽  
Vol 6 (5) ◽  
pp. 531-534 ◽  
Author(s):  
Kazuya Kato ◽  
Junji Kato ◽  
W. John B. Hodgson ◽  
Nader G. Abraham ◽  
Kazuhiko Onodera ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Cytochrome P450 ◽  
Enzymatic Activity ◽  
Microscopic Examination ◽  
Spontaneously Hypertensive Rats ◽  
Hypertensive Rats ◽  
Hydroxylase Activity ◽  
Spontaneously Hypertensive ◽  
Fetal Hepatocytes ◽  
Red Pulp

We examined the expression and enzymatic activity of the cytochrome P450 LAω within transplanted hepatocytes. Fetal hepatocytes were harvested at day 20 of gestation from spontaneously hypertensive rats (SHRs) and transplanted into recipient adult SHR spleens. Microscopic examination of the recipient spleens 4 and 10 wk after transplantation revealed masses of hepatocytes with cordlike structures in the red pulp. Immunochemical studies detected cytochrome (cyto) P450 LAω in the fetal hepatocytes before transplantation without prior induction. Although the cyto P450 LAω was not detected by the second week after transplantation, by the 6th and 10th wk after transplantation, it was. Cyto P450-arachidonic acid ω/4oM-1 hydroxylase activity (formation of 20- and 19-hydroxyeico-satetraenoic acid) was detected at 10 wk after transplantation, but not 2 or 6 wk after transplantation. These results demonstrated that fetal hepatocytes can be transplanted successfully into recipient spleens and then grow in the spleens, as in the case of the adult hepatocyte response.

scholarly journals Interrenal steroid 21-hydroxylase in eels: primary structure, progesterone-specific activity and enhanced expression by ACTH

2003 ◽  
Vol 31 (2) ◽  
pp. 327-340 ◽  
Author(s):  
YY Li ◽  
K Inoue ◽  
Y Takei
Keyword(s):  
Specific Activity ◽  
Head Kidney ◽  
Functional Expression ◽  
Amino Acid Identity ◽  
Mrna Levels ◽  
Hydroxylase Activity ◽  
Enhanced Expression ◽  
Key Enzyme ◽  
Steroid Binding ◽  
First Time

Cytochrome P450 21-hydroxylase (P450c21) is a key enzyme for corticosteroidogenesis. To understand the regulatory mechanisms of cortisol production in fish, we have cloned a cDNA encoding P450c21, for the first time in non-mammalian vertebrates, from the head kidney of the eel (Anguilla japonica). The overall similarity of the deduced P450c21 sequence was modest (41-44% amino acid identity) between the eel and mammals. However, the functional domains for steroid-binding, heme-binding and proton-transfer sites were well conserved (74-100% identity). The eel P450c21 mRNA was expressed abundantly in the anterior quarter of the head kidney, but was undetectable in the remaining three-quarters or in other tissues including the gill, heart, liver, intestine, kidney, immature gonad and skeletal muscle. Functional expression of the cDNA clone in non-steroidogenic COS-1 cells produced a protein with high 21-hydroxylase activity to convert progesterone to 11-deoxycortisterone but not 17alpha-hydroxyprogesterone to 11-deoxycortisol, although the latter is a preferred substrate for mammalian P450c21. To examine whether 21-hydroxylated progesterone is actually 17alpha-hydroxylated in the eel interrenal, 11-deoxycorticosterone and (3)H-corticosterone were respectively incubated with the interrenal-containing anterior quarter of the head kidney. The separation of the steroids produced by two HPLC systems revealed that cortisol was produced from both substrates, showing the 17alpha-hydroxylation of 11-deoxycorticosterone and corticosterone in the eel interrenal. ACTH infused at 3 pmol/kg per min for 5 h consistently increased plasma cortisol levels and interrenal P450c21 mRNA levels in seawater eels. These results showed that the interrenal-specific eel P450c21 cloned in this study is involved in cortisol production through conversion of progesterone to 11-deoxycorticosterone in the interrenal-containing anterior quarter of eel head kidney. Furthermore, ACTH stimulates cortisol production in part through enhanced P450c21 expression in the eel interrenal.

The cytochemical localization of alkaline phosphatase in the testicular nephridia of the Indian leech, Hirudinaria granulosa, with special reference to the inner lobe

1965 ◽  
Vol s3-106 (73) ◽  
pp. 31-35
Author(s):  
BHOOMITTRA DEV
Keyword(s):  
Alkaline Phosphatase ◽  
Enzymatic Activity ◽  
Special Reference ◽  
Functional Significance ◽  
Central Canal ◽  
Cytochemical Localization ◽  
First Time ◽  
Peripheral Cytoplasm

Previous studies by the author have revealed the intense localization of alkaline phosphatase in the initial-lobe cells and in the intracellular central canal on its first ‘round’ in the testicular nephridia of Hirudinaria granulosa. The present work throws light, for the first time, on the detailed cytochemical localization of the enzyme in the region where these two components communicate with each other. Openings of the central canal into an inner-lobe cell and of the latter into an initial-lobe cell have been noticed at one and the same level. An enzymatically negative inner-lobe cell serves as a bridge between the two positive zones. The bridge is, however, rarely observed because of the tendency to direct coalescence between the two enzymatically positive parts. The initial lobe sometimes opens into the central canal by means of more than one aperture. After the opening of the initial lobe into the central canal, the latter loses its enzymatic activity, and the radially arranged tubular inner-lobe cells disappear. The enzymatic activity in the peripheral cytoplasm of the intracellular central canal is usually more intense towards the point of its union with the initial lobe. The functional significance of the findings is discussed. Evidence for the concentration of the enzyme round the nucleus and in the nucleolus has also been noticed in the nephridial cells.

scholarly journals AN ENZYME IN MAST CELLS WITH PROPERTIES LIKE CHYMOTRYPSIN

1959 ◽  
Vol 110 (3) ◽  
pp. 451-460 ◽  
Author(s):  
Earl P. Benditt ◽  
Margaret Arase
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Methyl Ester ◽  
Enzymatic Activity ◽  
Differential Centrifugation ◽  
Cell Enzyme ◽  
Sucrose Solutions ◽  
Naphthoic Acid ◽  
Activity Curve ◽  
Chymotrypsin A

Mast cells contain an enzyme which hydrolyzes 3-chloroacetoxy-2-naphthoic acid anilide. By using highly purified mast cells isolated by differential centrifugation in high density sucrose solutions we have been able to study this enzymatic activity in more detail. The enzyme has properties similar to those of chymotrypsin: Chymotrypsin will hydrolyze the histochemical substrate, and the chymotrypsin and mast cell activities with this substrate are similarly inhibited by diisopropylfluorophosphate. The mast cell enzyme is capable of hydrolyzing the N-acetyl esters of tryptophan, tyrosine, and phenylalanine, the relative rates of hydrolysis being similar to those seen with chymotrypsin. A characteristic trypsin substrate, p-toluenesulfonyl arginine methyl ester, is not acted upon by the mast cell enzyme or chymotrypsin. The pH activity curve of the new cell enzyme is similar to that of chymotrypsin as determined with N-acetyl-L-tryptophan ethyl ester as substrate.

scholarly journals The Common P450 Oxidoreductase Variant A503V Is Not a Modifier Gene for 21-Hydroxylase Deficiency

2008 ◽  
Vol 93 (7) ◽  
pp. 2913-2916 ◽  
Author(s):  
Larissa G. Gomes ◽  
Ningwu Huang ◽  
Vishal Agrawal ◽  
Berenice B. Mendonça ◽  
Tania A. S. S. Bachega ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Maximum Velocity ◽  
Modifier Gene ◽  
Wild Type ◽  
Hydroxylase Activity ◽  
Michaelis Constant ◽  
Hydroxylase Deficiency ◽  
Cyp21a2 Gene ◽  
P450 Oxidoreductase ◽  
21 Hydroxylase Deficiency

Abstract Context: 21-hydroxylase deficiency (21OHD) is a common genetic disorder caused by mutations in the CYP21A2 gene, which encodes the adrenal 21-hydroxylase, microsomal P450c21. CYP21A2 gene mutations generally correlate well with impaired P450c21 enzymatic activity and the clinical findings in 21OHD, but occasional discrepancies between genotype and phenotype suggest the effects of modifier genes. Mutations in P450 oxidoreductase (POR), the protein that transfers electrons from reduced nicotinamide adenine dinucleotide phosphate to all microsomal P450s, can ameliorate the 21OHD phenotype and, therefore, could be a modifier gene. Objectives: We sought to identify POR variants in patients with 21OHD having discordant phenotype and genotype, and to evaluate their effect on 21-hydroxylase activity. Patients and Methods: We determined the CYP21A2 genotypes of 313 Brazilian patients with 21OHD and correlated the genotype and phenotype. The POR gene was sequenced in 17 patients with discordant genotype and phenotype. Wild-type and A503V POR, and P450c21 were expressed in bacteria and reconstituted in vitro. Activities were assayed by conversion of [14C]progesterone to deoxycorticosterone and [3H]17-hydroxyprogesterone to 11-deoxycortisol, and assessed by thin layer chromatography and phosphorimaging. Results: The A503V POR variant was found in 10 of 30 alleles, the same ratio as in the normal population. There were no significant differences in Michaelis constant, maximum velocity and maximum velocity/Michaelis constant of 21-hydroxylase activity supported by wild-type and A503V POR. Conclusion: The only POR missense polymorphism found in atypical 21OHD patients was A503V. Although A503V reduces P450c17 enzymatic activity, it does not influence P450c21 activity, indicating that POR A503V does not modify the 21OHD phenotype.

Immobilization of Bacillus subtilis lipase on a Cu-BTC based hierarchically porous metal–organic framework material: a biocatalyst for esterification

2016 ◽  
Vol 45 (16) ◽  
pp. 6998-7003 ◽  
Author(s):  
Yu Cao ◽  
Zhuofu Wu ◽  
Tao Wang ◽  
Yu Xiao ◽  
Qisheng Huo ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Enzymatic Activity ◽  
Metal Organic Framework ◽  
Porous Metal ◽  
Esterification Reaction ◽  
Hierarchically Porous ◽  
Metal Organic ◽  
First Time ◽  
High Enzymatic Activity ◽  
Organic Framework

Bacillus subtilislipase (BSL2) has been successfully immobilized into a Cu-BTC based hierarchically porous MOF material for the first time. The immobilized BSL2 presents high enzymatic activity and perfect reusability during the esterification reaction.

scholarly journals Sugarcane Bagasse Saccharification by Enzymatic Hydrolysis Using Endocellulase and β-glucosidase Immobilized on Different Supports

Catalysts ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 340
Author(s):  
Wilson Morais Junior ◽  
Thályta Pacheco ◽  
Shipeng Gao ◽  
Pedro Martins ◽  
José Guisán ◽  
...  
Keyword(s):  
Enzymatic Hydrolysis ◽  
Enzymatic Activity ◽  
Sugarcane Bagasse ◽  
Commercial Enzymes ◽  
Iron Oxide Magnetic Nanoparticles ◽  
Pretreated Bagasse ◽  
Pretreated Sugarcane Bagasse ◽  
Aldehyde Groups ◽  
Solid Liquid ◽  
First Time

The saccharification of sugarcane bagasse by enzymatic hydrolysis is one of the most promising processes for obtaining fermentable sugar to be used in the production of second-generation ethanol. The objective of this work was to study the immobilization and stabilization of two commercial enzymes: Endocellulase (E-CELBA) in dextran coated iron oxide magnetic nanoparticles activated with aldehyde groups (DIOMNP) and β-glucosidase (E-BGOSPC) in glyoxyl agarose (GLA) so that their immobilized derivatives could be applied in the saccharification of pretreated sugarcane bagasse. This was the first time that the pretreated sugarcane bagasse was saccharified by cascade reaction using a endocellulase immobilized on dextran coated Fe2O3 with aldehyde groups combined with a β-glucosidase immobilized on glyoxyl agarose. Both enzymes were successfully immobilized (more than 60% after reduction with sodium borohydride) and presented higher thermal stability than free enzymes at 60, 70, and 80 °C. The enzymatic hydrolysis of the sugarcane bagasse was carried out with 15 U of each enzyme per gram of bagasse in a solid-liquid ratio of 1:20 for 48 h at 50 °C. Under these conditions, 39.06 ± 1.18% of the cellulose present in the pretreated bagasse was hydrolyzed, producing 14.11 ± 0.47 g/L of reducing sugars (94.54% glucose). In addition, DIOMNP endo-cellulase derivative maintained 61.40 ± 1.17% of its enzymatic activity after seven reuse cycles, and GLA β-glucosidase derivative maintained up to 58.20 ± 1.55% of its enzymatic activity after nine reuse cycles.

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