scholarly journals Isolation and characterization of mitochondria and lysosome from isoproterenol induced cardiotoxic rats

2019 ◽  
Vol 8 (1) ◽  
pp. 12-18
Author(s):  
Malathi Mangalanathan ◽  
Tamiloli Devendhiran ◽  
Saraswathi Uthamaramasamy ◽  
Keerthika Kumarasamy ◽  
K Mohanraj ◽  
...  
Keyword(s):  
Heart Tissue ◽  
Cardiac Cells ◽  
Marker Enzymes ◽  
Membrane Function ◽  
Biochemical Alterations ◽  
Isolation And Characterization ◽  
The Stability ◽  
Membrane Pretreatment ◽  
Hydroethanolic Extract

Mitochondrial and lysosomal membranes are prominent membranes of cardiac cells and are the factors that determine membrane function in myocardial ischemia. In this study, isolation of mitochondria and lysosome from heart tissue under the control, isoproterenol (ISO) (8.5mg/100g) induced cardiotoxic rats and oral pretreatment with Z. armatum fruit (200, 400mg/kg body weight) treated rats. Further characterization of marker enzymes was done. A decreased in the activity of all the mitochondrial and lysosomal marker enzymes in ISO administered cardiotoxic rats when compared to control rats which indicate ISO decreased the stability of the membrane. Pretreatment with hydroethanolic extract of Z. armatum fruit to ISO induced rats significantly reverted these biochemical alterations near to normal. The possible mechanism for the protection of heart mitochondria and lysosome against oxidative damage induced by ISO might be due to quenching of free radicals and enhancing the action of marker enzymes.

scholarly journals Isolation and characterization of rabbit kidney brush borders

1972 ◽  
Vol 128 (5) ◽  
pp. 1319-1328 ◽  
Author(s):  
S. J. Quirk ◽  
G. B. Robinson
Keyword(s):  
Plasma Membranes ◽  
Sucrose Density Gradient ◽  
Molar Ratio ◽  
Rabbit Kidney ◽  
Kidney Cortex ◽  
Marker Enzymes ◽  
Isolation And Characterization ◽  
Adenosine Triphosphatases ◽  
Brush Borders

1. Brush borders were isolated from rabbit kidney-cortex homogenates by rate-zonal centrifugation through a sucrose density gradient in a B-XIV zonal rotor, followed by differential centrifugation. 2. The method of preparation gave brush borders of high purity with a reasonable yield. The morphological appearance supported the evidence from enzymic and chemical investigations, that the brush borders were only slightly contaminated with endoplasmic reticulum, mitochondria, lysosomes and nuclei. 3. The molar ratio of cholesterol to phospholipid lay within the range found in other plasma membranes, but the carbohydrate content was double that found in liver plasma membranes. 4. Alkaline phosphatase, maltase, trehalase and aminopeptidase were major enzymic constituents of the brush borders, and had an approximately equal yield and enrichment, but none of these enzymes fulfilled the criteria for marker enzymes. 5. Mg2+-dependent and Na+,K+-dependent adenosine triphosphatases, although found in brush borders, had low yields and low enrichments.

scholarly journals Produksi, isolasi dan karakterisasi superoksida dismutase dari Spirulina platensis yang dibiakkan dalam serum lateks Production, isolation, and characterization of superoxyde dismutase from Spirulina platensis cultured on latex serum

2016 ◽  
Vol 77 (1) ◽  
Author(s):  
. TRI-PANJI ◽  
. SUHARYANTO ◽  
Marini WIJAYANTI
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Spray Drying ◽  
Spirulina Platensis ◽  
Active Agent ◽  
Pilot Scale ◽  
Isolation And Characterization ◽  
The Stability ◽  
Major Amino Acid

AbstractSpirulina platensis is a blue-green microalgawhich is frequently used for food and feedsupplements and cosmetic active agent. Thismicroalga also produces a strong antioxidantnamely superoxide dismutase (SOD) used ascosmetic active agent for anti aging and anti freeradicals. SOD was isolated from S. platensis cellbiomass from local isolate grown on latex serumon semipilot (3.5 m 3 ) and pilot scale (40 m 3 )then dried with spray drying or sun drying andcharacterized. SOD was purified with sequentialtwo-stage sedimentation using ammoniumsulphate and fractionated in chromatographiccolumn containing Sephadex G 200. Thefractions were analysed to determine the activity,cofactor metal and amino acid composition of theantioxidant. The results showed thatsedimentation of SOD extract with 80%ammonium sulphate produced SOD with higheractivity compared to that of SOD fromcommercial S. platensis biomass. This SOD wassuccessfully isolated and purified. MetaloenzymeSOD was composed of subunits with molecularweight of 77.78; 71.74; and 19.2 kDa, whichcontained nine types of amino acids with tyrosineand lysine as the major amino acid components.Zn was the most predominant metal on SOD, thenfollowed by Fe and Mn. The main subunitcofactors consisted of Zn 72%, Fe 25%, Mn 2%,and Cu 1%, which were different from thesmall subunit that contained of Zn 55%, Mn 31%,Fe 14%, and Cu 4%. The stability of SOD wasachieved on pH 7.5 and temperature below 25 o C.AbstrakSpirulina platensis adalah mikroalga hijaubiru yang banyak digunakan sebagai suplemenpangan, pakan, dan bahan aktif kosmetika.Mikroalga ini juga menghasilkan antioksidankuat yaitu superoksida dismutase (SOD), yangmerupakan bahan aktif kosmetika anti penuaandini dan pencegah efek radikal bebas. SODdiisolasi dari biomassa sel S. platensis isolat lokalyang dibiakkan dalam serum lateks skalasemipilot (3,5 m 3 ) dan pilot (40 m 3 ) sertadikeringkan dengan cara pengeringan kabut(spray drying) atau penjemuran untuk kemudiandikarakterisasi. SOD dimurnikan dengan peng-endapan bertingkat menggunakan ammoniumsulfat dan dipisahkan dengan kolom kromatografiberisi Sephadex G 200. Hasil pemisahankemudian dianalisis untuk menentukan aktivitas,logam kofaktor serta komposisi asam amino antioksidan tersebut. Hasil penelitian menunjukkanbahwa pengendapan ekstrak SOD denganSOD lebih tinggi dari SOD asal biomassaS. platensis komersial. SOD tersebut telahberhasil diisolasi dan dimurnikan. MetaloenzimSOD tersusun atas subunit dengan BM 77,78;71,74; dan 19,2 kDa, yang mengandungsembilan jenis asam amino dengan tirosin danlisin sebagai komponen asam amino utama.Logam yang dominan pada SOD adalah Zn,disusul kemudian Fe dan Mn. Kofaktor sub unitbesar terdiri dari Zn 72%, Fe 25%, Mn 2%, danCu 1%, berbeda dengan sub unit kecil yangmengandung Zn 55%, Mn 31%, Fe 14%, dan Cu4%. Stabilitas SOD S. platensis dicapai pada pH7,5 dan suhu di bawah 25 o Cammonium sulfat 80% menghasilkan aktivitas

Isolierung und Charakterisierung einer Acetylester-Hydrolase aus Aspergillus rtiger / Isolation and Characterization of an Acetylester-Hydrolase from Aspergillus niger

1980 ◽  
Vol 35 (9-10) ◽  
pp. 696-698 ◽  
Author(s):  
B. Schöbel ◽  
W. Pollmann
Keyword(s):  
Molecular Weight ◽  
Acetic Acid ◽  
Temperature Optimum ◽  
Isolation And Characterization ◽  
Glyceryl Triacetate ◽  
Characteristic Features ◽  
The Stability ◽  
Acetic Acid Esters ◽  
Acid Esters

Abstract The characteristic features of an acetic acid esters hydrolyzing enzyme (acetylesterase, EC 3.1.1.16) are described. The pH- and temperature optimum were 7.0 and 40 °C respectively. The stability of the enzyme regarding different pH- and temperature conditions was investigated. The molecular weight of the acetylesterase could be determined to 160000. A small acetic ester hydrolyzing activity was found too with a molecular weight of about 25000. The activity was not inhibited by addition of di-isopropylphosphorofluoridate (DFP) or physostigmine. The KM-value for glyceryl triacetate was about 90 mM. Concentration of the enzyme was done by ultrafiltration and column-chromatography. The enzymatic activity tests were performed titrimetrically using glyceryl triacetate for substrate.

One-step isolation and characterization of nuclear membranes

1974 ◽  
Vol 268 (891) ◽  
pp. 101-108 ◽  
Keyword(s):  
Rat Liver ◽  
Concanavalin A ◽  
Nuclear Membrane ◽  
Marker Enzymes ◽  
Nuclear Membranes ◽  
Isolation And Characterization ◽  
One Step ◽  
Freeze Etching

Purified fractions of nuclei from rat liver and isolated calf thymocytes are treated with 0.5 M MgCl 2 or with an alkaline buffer, pH 8.5. Such procedures dissociate nuclear membranes from the chromatin. The membranes are simply floated in a gradient. The recovery in the membrane band obtained by the Mg 2+ technique is 55 to 60 %. Freeze-etching studies of the membrane pellets provide a good way to check their purity, pores being very conspicuous. A 5'-nucleotidase and an ATPase are detected on the nuclear membranes of thymocytes, as well as a still undescribed 3'-nucleotidase. However, no marker enzymes for the nuclear membrane have been found. The topology of sugar residues on the nuclear membranes is studied by means of lectins. Concanavalin A binds in large amounts to nuclear envelopes, both on the inner and outer leaflets, but not to pores.

scholarly journals Isolation and characterization of pigeon breast muscle cytosolic 5'-nucleotidase-I (cN-I).

2005 ◽  
Vol 52 (4) ◽  
pp. 789-796 ◽  
Author(s):  
Kinga Tkacz-Stachowska ◽  
Katarzyna Lechward ◽  
Andrzej C Skladanowski
Keyword(s):  
Physiological Function ◽  
Specific Activity ◽  
Heart Tissue ◽  
Breast Muscle ◽  
Kinetic Properties ◽  
Type I ◽  
Isolation And Characterization ◽  
Atp Breakdown ◽  
Wet Weight

5'-Nucleotidase specific towards dCMP and AMP was isolated from avian breast muscle and characterized. It was found to be similar to a type-I form (cN-I) identified earlier as the AMP-selective 5'-nucleotidase responsible for adenosine formation during ATP breakdown in transfected COS-7 cells. Expression pattern of the cN-I gene in pigeon tissues indicated breast muscle as a rich source of the transcript. We purified the enzyme from this source using two-step chromatography and obtained an active homogenous preparation, free of ecto-5'-nucleotidase activity. The tissue content of the activity was calculated at 0.09 U/g wet weight. The specific activity of the enzyme preparation was 4.33 U/mg protein and it preferred dCMP and AMP to dAMP and IMP as a substrate. Its kinetic properties were very similar to those of the enzyme purified earlier from heart tissue. It was strongly activated by ADP. Inhibition by inorganic phosphate was more pronounced than in heart-isolated cN-I. Despite this difference, a similar physiological function is suggested for cN-I in both types of muscle.

scholarly journals Produksi, isolasi dan karakterisasi superoksida dismutase dari Spirulina platensis yang dibiakkan dalam serum lateks Production, isolation, and characterization of superoxyde dismutase from Spirulina platensis cultured on latex serum

2016 ◽  
Vol 77 (1) ◽  
Author(s):  
. TRI-PANJI ◽  
. SUHARYANTO ◽  
Marini WIJAYANTI
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Spray Drying ◽  
Spirulina Platensis ◽  
Active Agent ◽  
Pilot Scale ◽  
Isolation And Characterization ◽  
The Stability ◽  
Major Amino Acid

AbstractSpirulina platensis is a blue-green microalgawhich is frequently used for food and feedsupplements and cosmetic active agent. Thismicroalga also produces a strong antioxidantnamely superoxide dismutase (SOD) used ascosmetic active agent for anti aging and anti freeradicals. SOD was isolated from S. platensis cellbiomass from local isolate grown on latex serumon semipilot (3.5 m 3 ) and pilot scale (40 m 3 )then dried with spray drying or sun drying andcharacterized. SOD was purified with sequentialtwo-stage sedimentation using ammoniumsulphate and fractionated in chromatographiccolumn containing Sephadex G 200. Thefractions were analysed to determine the activity,cofactor metal and amino acid composition of theantioxidant. The results showed thatsedimentation of SOD extract with 80%ammonium sulphate produced SOD with higheractivity compared to that of SOD fromcommercial S. platensis biomass. This SOD wassuccessfully isolated and purified. MetaloenzymeSOD was composed of subunits with molecularweight of 77.78; 71.74; and 19.2 kDa, whichcontained nine types of amino acids with tyrosineand lysine as the major amino acid components.Zn was the most predominant metal on SOD, thenfollowed by Fe and Mn. The main subunitcofactors consisted of Zn 72%, Fe 25%, Mn 2%,and Cu 1%, which were different from thesmall subunit that contained of Zn 55%, Mn 31%,Fe 14%, and Cu 4%. The stability of SOD wasachieved on pH 7.5 and temperature below 25 o C.AbstrakSpirulina platensis adalah mikroalga hijaubiru yang banyak digunakan sebagai suplemenpangan, pakan, dan bahan aktif kosmetika.Mikroalga ini juga menghasilkan antioksidankuat yaitu superoksida dismutase (SOD), yangmerupakan bahan aktif kosmetika anti penuaandini dan pencegah efek radikal bebas. SODdiisolasi dari biomassa sel S. platensis isolat lokalyang dibiakkan dalam serum lateks skalasemipilot (3,5 m 3 ) dan pilot (40 m 3 ) sertadikeringkan dengan cara pengeringan kabut(spray drying) atau penjemuran untuk kemudiandikarakterisasi. SOD dimurnikan dengan peng-endapan bertingkat menggunakan ammoniumsulfat dan dipisahkan dengan kolom kromatografiberisi Sephadex G 200. Hasil pemisahankemudian dianalisis untuk menentukan aktivitas,logam kofaktor serta komposisi asam amino antioksidan tersebut. Hasil penelitian menunjukkanbahwa pengendapan ekstrak SOD denganSOD lebih tinggi dari SOD asal biomassaS. platensis komersial. SOD tersebut telahberhasil diisolasi dan dimurnikan. MetaloenzimSOD tersusun atas subunit dengan BM 77,78;71,74; dan 19,2 kDa, yang mengandungsembilan jenis asam amino dengan tirosin danlisin sebagai komponen asam amino utama.Logam yang dominan pada SOD adalah Zn,disusul kemudian Fe dan Mn. Kofaktor sub unitbesar terdiri dari Zn 72%, Fe 25%, Mn 2%, danCu 1%, berbeda dengan sub unit kecil yangmengandung Zn 55%, Mn 31%, Fe 14%, dan Cu4%. Stabilitas SOD S. platensis dicapai pada pH7,5 dan suhu di bawah 25 o Cammonium sulfat 80% menghasilkan aktivitas

scholarly journals Isolation and characterization of a Sca-1+/CD31- progenitor cell lineage derived from mouse heart tissue

2014 ◽  
Vol 14 (1) ◽  
pp. 75 ◽  
Author(s):  
Hao Wang ◽  
Hao Chen ◽  
Bei Feng ◽  
Xiang Wang ◽  
Xiaomin He ◽  
...  
Keyword(s):  
Progenitor Cell ◽  
Cell Lineage ◽  
Heart Tissue ◽  
Mouse Heart ◽  
Isolation And Characterization
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