scholarly journals Characterization of Proteolytic and Collagenolytic Enzymes from the Larvae of Lucilia cuprina, the Sheep Blowfly

1988 ◽  
Vol 41 (2) ◽  
pp. 269 ◽  
Author(s):  
VM Bowles ◽  
PR Carnegie ◽  
RM Sandeman
Keyword(s):  
Ethylenediaminetetraacetic Acid ◽  
Proteolytic Enzymes ◽  
Chelating Agent ◽  
Collagenolytic Activity ◽  
Enzyme Preparations ◽  
Molecular Weights ◽  
Highly Active ◽  
Wide Range ◽  
Secretory Enzyme

Isoelectric focusing was used to characterize proteolytic enzymes in homogenate and excretory-secretory preparations of the larvae of L. cuprina, the sheep blowfly. Zymogram overlays showed that the larvae produce a number of highly active proteases which have a wide range of isoelectric points and molecular weights. The alkaline and neutral pI proteases were inhibited by phenylmethyl-sulfonylfluoride, leupeptin and aprotinin; this indicated the presence of serine in the active site. Pepstatin and the metal chelating agent ethylenediaminetetraacetic acid had no effect Oil the activity of any of the proteases. Optimal pH for activity of the proteases was between 7 and 8. In addition, the proteases were found to be heat labile. Digestion of collagen fibrils confirmed the existence of collagenolytic activity in the excretory-secretory enzyme preparations. It is suggested that these enzymes may be involved in the nutrition of the larvae and in the pathogenesis of the lesion on the skin.

Synthesis and Characterization of Strontium-Doped Lanthanum Cobaltite Cathode Material Prepared by a Modified Combined Complexing Method

2013 ◽  
Vol 701 ◽  
pp. 131-135 ◽  
Author(s):  
Abdullah Abdul Samat ◽  
Mohd Azlan Mohd Ishak ◽  
Hamidi Abd Hamid ◽  
Osman Nafisah
Keyword(s):  
Cathode Material ◽  
Ethylenediaminetetraacetic Acid ◽  
Perovskite Phase ◽  
Chelating Agent ◽  
Xrd Analysis ◽  
Bulk Sample ◽  
Potential Candidate ◽  
X Ray Diffraction ◽  
Lanthanum Cobaltite

A potential candidate for intermediate temperature solid oxide fuel cell (IT-SOFC) cathode material which is strontium-doped lanthanum cobaltite, La0.6Sr0.4CoO3-α (LSCO) has been synthesized by a complexing method. Citric acid (CA) and ethylenediaminetetraacetic acid (EDTA) were used as a combined chelating agent and ethylene glycol (EG) was employed as surfactant. The obtained powder was characterized by thermogravimetric analysis (TGA), X-ray diffraction (XRD) analysis and scanning electron microscopy (SEM). TGA results showed the thermal decomposition of the precursor gel was completed at 700 °C. A single perovskite phase of LSCO with cubic structure was obtained at calcination temperature of 1000 °C with heating/cooling rate of 10 °C min-1 as confirmed by XRD analysis. SEM result revealed that the morphology of the powder was spherical in shape with diameter ranging from 250 to 650 nm. Apparently, the bulk sample consists of almost homogeneous and identical particles.

Highly active and stable protease production by an extreme halophilic archaeon Haloarcula sp. TG1 isolated from Lake Tuz, Turkey

2017 ◽  
Vol 42 (3) ◽  
Author(s):  
Büşra Abanoz ◽  
Sezer Okay ◽  
Aslıhan Kurt-Kızıldoğan
Keyword(s):  
Protease Activity ◽  
Protease Production ◽  
Extreme Conditions ◽  
Salt Mine ◽  
Producer Strain ◽  
Extreme Halophiles ◽  
Highly Active ◽  
Protease Enzyme ◽  
Wide Range

AbstractObjective:Isolation of halophilic microorganisms from Çankırı salt mine and Lake Tuz in Turkey to explore versatile protease producers for industry and characterization of protease enzyme from the best protease producer among the isolated strains.Methods:Extreme halophiles were isolated from salt samples of Çankırı salt mine and Lake Tuz. Their protease activities were determined. The isolate with the highest protease activity was characterized. Its protease activity was evaluated in different NaCl concentrations, temperature and pH ranges, and in the presence of different inhibitors and metals. Thermostability and pH stability were also determined.Results:The highest protease producer strain was identified asConclusion:Due to the high activity and stability over a wide range of extreme conditions,

scholarly journals Characterization of proteins structurally related to human N-acetyl-β-d-glucosaminidase

1978 ◽  
Vol 173 (1) ◽  
pp. 191-196 ◽  
Author(s):  
M Carroll
Keyword(s):  
Molecular Weight ◽  
Human Liver ◽  
Deae Cellulose ◽  
Low Molecular Weight ◽  
Enzyme Preparations ◽  
Molecular Weights ◽  
Functional Relationships ◽  
Hexosaminidase A ◽  
Cellulose Column

Those proteins of human liver that cross-reacted with antibodies raised to apparently homogenous hexosamindases A and B were detected by immunodiffusion. Cross-reacting proteins with high molecular weights (greater than 2000000) and intermediate molecular weights (70000–200000) were present both in the unadsorbed fraction and in the 0.05–0.2M-NaCl eluate obtained by DEAE-cellulose chromatography at pH7.0. The unadsorbed fraction also contained a cross-reacting protein of low molecular weight (10000–70000). The possible structural and functional relationships between hexosaminidase and the cross-reacting proteins are discussed. An apparently cross-reacting protein present in the 0.05M-NaCl eluate from the DEAE-cellulose column was serologically unrelated to hexosaminidase, but it gave a reaction of immunological identify with one of the apparently cross-reacting proteins having the charge and size characteristics of hexosaminidase A. It is suggested that immunochemical methods may provide criteria for the homogeneity of enzyme preparations superior to those of conventional methods.

scholarly journals Purification and Partial Characterization of Rat Fibrinogen (rat-Fbg)

1977 ◽  
Author(s):  
W. Nieuwenhuizen ◽  
Irina A.M. van Ruijven-Vermeer
Keyword(s):  
Amino Acid ◽  
Gel Electrophoresis ◽  
Proteolytic Enzymes ◽  
Mammalian Species ◽  
Rat Plasma ◽  
Gel Chromatography ◽  
Molecular Weights ◽  
Ammonium Sulphate Precipitation ◽  
Proteolytic Activities

Rat-Fbg was purified from rat plasma using Sepharose-1ysi ne chromatography, repeated ammonium sulphate precipitation (35% saturation) and gel chromatography on Sepharose 6B.In order to minimize proteolytic activity, rats were injected intravenously with Trasylol before bleeding, and the collected blood was treated with Trasylol and DFP.A preparation was obtained, which was 95% clottable and showed a single band on SDS-poly-acrylamide gel electrophoresis. Alanine was the only detectable am i no-term i na 1 amino acid.After reduction and modification of the SH groups the material could be separated into three distinct chains (Aα, Bβ and γ) by pore-limit Polyacrylamide slab-gel electrophoresis in sod i urn dodecyl sulphate. The amino acid composition of the whole Fbg and of the separated modified chains were determined. The molecular weights were 61,000, 58,000 and 51,000 for Aα, Bβ and γ-chains, respectively.In as far as the chains are concerned, our results are in contrast with the findings of Bouma et al. (J. Biol. Chem. 250(1975) 4678), who could not discriminate between Aα- and Bβ-chains in SDS-polyacry1 am i de gel electrophoresis. Evidence will be presented that this can be due to Aa-chain degradation caused by incomplete inhibition of proteolytic enzymes during the purification.It is concluded, that complete inhibition of proteolytic activities in all purification steps is essential to obtain native fibrinogen. Moreover, in contrast to the conclusions of Bouma rat-Fbg does not differ essentially from Fbg from other mammalian species.

scholarly journals Production and Characterization of Alkaline Protease from a High Yielding and Moderately Halophilic Strain of SD11 Marine Bacteria

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Hongxia Cui ◽  
Liping Wang ◽  
Yang Yu
Keyword(s):  
Alkaline Protease ◽  
Marine Bacterium ◽  
Ethylenediaminetetraacetic Acid ◽  
Agar Plate ◽  
Skim Milk ◽  
Optimal Temperature ◽  
Phenylmethylsulfonyl Fluoride ◽  
Highly Active ◽  
Sea Area

A marine bacterium SD11, which was isolated from sea muds (Geziwo Qinhuangdao Sea area, China), was used to produce thermostable alkaline serine nonmetal protease in the skim milk agar plate medium with 10% NaCl. The optimal temperature about the manufacture of the extracellular protease was ~60°C. The crude enzyme was stable at 20–50°C. The activity was retained to 60% and 45% after heating for 1 h at 60 and 70°C, respectively. The protease was highly active in a wide pH scope (8.0–10.0) and maximum protease activity exhibited at pH 10.0. The activity was restrained by phenylmethylsulfonyl fluoride (PMSF) but mildly increased (~107%) in the presence of ethylenediaminetetraacetic acid (EDTA), indicating that the production contains serine-protease(s) and nonmetal protease(s). Moreover, the crude alkaline protease was active with the 5 mM Ca2+, Mn2+, Zn2+, Cu2+, Na+, and K+that existed separately. In addition, the protease showed superduper stability when exposed to an anionic surfactant (5 mM SDS), an oxidizing agent (1% H2O2), and several organic solvents (methanol, isopropanol, and acetone). These results suggest that the marine bacterium SD11 is significant in the industry from the prospects of its ability to produce thermally stable alkaline protease.

Purine catabolism in man: characterization of placental microsomal 5′-nucleotidase

1976 ◽  
Vol 54 (5) ◽  
pp. 462-469 ◽  
Author(s):  
Irving H. Fox ◽  
Pamela J. Marchant
Keyword(s):  
Specific Activity ◽  
Ph Optimum ◽  
Electrophoretic Variants ◽  
Molecular Weights ◽  
Wide Range ◽  
Stokes Radius ◽  
Michaelis Constants ◽  
Light Form ◽  
Molecular Weight Form

Human placental microsomal 5′-nucleotidase (EC 3.1.3.5) was prepared free of alkaline phosphatase by isoelectric focusing. A total of seven electrophoretic variants were isolated during the preparation of six placentas. Only three to six variants were found in a single placenta. The isoelectric pH's were 6.70, 6.44, 6.23, 6.02, 5.76, 5.63 and 5.44. These were found to be composed of variable quantities of a large, medium and low molecular weight form. The apparent molecular weights of the medium and light form of the enzyme were 86 500 and 43 500, respectively, as estimated from Stokes radius and sedimentation velocity determinations. The electrophoretic variants were not distinguishable with respect to specific activity and Michaelis constants for AMP, GMP or CMP or inhibition by ATP, CTP or adenosine. These electrophoretic variants appeared to be pseudoisozymes based upon different states of aggregation of a common primary sequence.There was a wide range of substrate specificity among nucleoside 5′-monophosphates which included 2-deoxyribose compounds. With AMP as 100, substrate activity was: CMP, 122; NMN, 74; GMP, 68; IMP, 63; XMP, 28 and UDP–glucose, 68. The Michaelis constants for AMP, GMP and CMP ranged from 12–18 μM, from 33–67 μM and from 170–250 μM, respectively. Although 5′-nucleotidase was active in the absence of divalent cation, 5 mM MgCl2 stimulated the enzyme activity to 234% of control and shifted the pH optimum of 9.8 to a plateau from pH 7.4–9.8.

scholarly journals SCREENING OF ASPERGILLUS FUNGI FOR EXTRA CELLULAR PROTEASE AND COLLAGENASE PRODUCTION

REPORTS ◽  
2020 ◽  
Vol 2 (330) ◽  
pp. 36-40
Author(s):  
R.K. Blieva ◽  
A.K. Kalieva ◽  
Zh.B. Suleimenova ◽  
A.S. Zhakipbekova ◽  
I.E. Tapenbayeva
Keyword(s):  
Proteolytic Enzymes ◽  
Raw Materials ◽  
Meat Industry ◽  
Improve Quality ◽  
Meat Processing ◽  
Enzyme Preparations ◽  
Technological Processes ◽  
Fungal Association ◽  
Selection Of

Protease and collagenase are the most important enzymes used for the processing of meat raw materials. In the meat industry, proteolytic enzymes are used to accelerate the maturation of meat and increase its yield. The use of enzyme preparations in meat processing makes it possible to rationally use meat raw materials, intensify technological processes, improve quality and expand the range of products. Collagenase, unlike protease, acts on those connective proteins of meat raw materials that determine its stiffness, breaking down hard-hydrolyzable and non-digestible collagen. The aim of this study was selection of strains of industrially valuable micromycetes from the collection of micromycetes that have the ability to synthesize extracellular protease and collagenase and create a fungal association. A comparative characterization of 7 strains of micromycetes of the genus Aspergillus and Penicillium - potential producers of protease and collagenase enzymes, was carried out. A. awamori 16 and A. awamori 22 showed the highest clearance zones and was used for further studies. The clearance zones of casein of A. awamori 16 on day 5 were 22.8 mm, and collagen 20.8 mm, while the clearance zones of casein of A. awamori 22 were 20.1 mm, and collagen - 19.1 mm.

Androgen-binding proteins in sheep epididymis: characterization of a cytoplasmic androgen receptor in the ram epididymis

1984 ◽  
Vol 103 (3) ◽  
pp. 273-279 ◽  
Author(s):  
S. Carreau ◽  
M. A. Drosdowsky ◽  
M. Courot
Keyword(s):  
Androgen Receptor ◽  
Binding Sites ◽  
Sucrose Gradient ◽  
Proteolytic Enzymes ◽  
Molecular Weights ◽  
High Affinity ◽  
Isoelectric Points ◽  
Extracellular Transport ◽  
Androgen Binding

ABSTRACT An androgen receptor (Rc) was demonstrated in caput, corpus and cauda epididymal cytosols of the ram. This receptor had a high affinity for 5α-dihydrotestosterone (Kd = 5·2 × 10−9 mol/l) and could be distinguished from the androgen-binding protein (ABP) by several characteristics. On polyacrylamidegel electrophoresis, Rc had a mobility of 0·37 and ABP 0·61; Rc sedimented in the 9S region of a linear sucrose gradient whereas ABP migrated in the 4·3S region; the molecular weights were 192 000 and 90 000 for Rc and ABP; their isoelectric points were 5·7 and 4·8–5·0; they were proteinaceous components since they were destroyed by proteolytic enzymes and heating (50 °C for Rc and 60 °C for ABP); they exhibited different half-times of dissociation: 20 h at 0 °C for Rc and 6 min for ABP, which is in agreement with their respective physiological roles, intra- and extracellular transport of androgens. The content of Rc-binding sites in caput epididymis was 18, in corpus 4 and in cauda 22 fmol/mg protein. J. Endocr. (1984) 103, 273–279

scholarly journals Evaluation of Factors Affecting Antimicrobial Activity of Bacteriocin from Lactobacillus plantarum Microencapsulated in Alginate-Gelatin Capsules and Its Application on Pork Meat as a Bio-Preservative

2019 ◽  
Vol 16 (6) ◽  
pp. 1017 ◽  
Author(s):  
Ngoc Le ◽  
Long Bach ◽  
Duy Nguyen ◽  
Tran Le ◽  
Khanh Pham ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Lactobacillus Plantarum ◽  
Incubation Temperature ◽  
Ethylenediaminetetraacetic Acid ◽  
Sodium Dodecyl ◽  
Fermented Foods ◽  
Pork Meat ◽  
Indicator Organisms ◽  
Highly Active ◽  
Wide Range

Antimicrobial compounds from traditional fermented foods have shown activity against a wide range of pathogen and spoilage microorganisms for several years. In this study, a Lactic acid bacteria (LAB), isolated from Vietnamese traditional fermented yogurt (Lactobacillus plantarum SC01), was encapsulated in alginate-gelatin (ALG-GEL) and the effect of incubation temperature, medium pH and surfactants were assessed. The aims of this research were to evaluate antimicrobial activity of bacteriocin produced by L. plantarum SC01. Another aim the research was to study the quality of pork meat treated with its Bacteriocin in 2 h as a bio-preservative at different storage times (0 h, 12 h, 24 h and 48 h) in room temperature, compared to control (treated with salt 40.0%). The antimicrobial activity of L. plantarum SC01 was identified through the inhibition rate of five indicator organisms, including Escherichia coli, Salmonella sp., Staphylococcus aureus, Listeria monocytogenes, and Bacillus subtilis by co-culture method. The results showed that L. plantarum SC01 microencapsulated in ALG-GEL (2.5% alginate and 6.0% gelatin, w/v) and 3.0% bacteria supplied into modified MRS medium (MRSOPTSC01) produced highly active compound inhibited the growth of indicator organisms at a density of 104–108 CFU/mL. Antibacterial compounds were highly active in a treatment at 80 °C; not to be affected by pH; affected by surfactant as Ethylenediaminetetraacetic acid (EDTA), Sodium dodecyl sulfate (SDS), and Tween. Moreover, LAB obtained from this study show the potent Bacteriocin in its usage as a preservative in food.

Purification and Characterization of an Alkaline Protease from Bacillus alcalophilus

2012 ◽  
Vol 192 ◽  
pp. 285-288 ◽  
Author(s):  
Shan Shan Liu ◽  
Li Li Wang ◽  
Lin Yuan
Keyword(s):  
Alkaline Protease ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Ammonium Sulfate Precipitation ◽  
Purification And Characterization ◽  
Highly Active ◽  
Wide Range ◽  
Commercial Applications ◽  
Ph Range

The purification and characterization of an alkaline protease produced by Bacillus alcalophilus were investigated. The enzyme was purified in two steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by cation-exchange chromatography. The purified protease had a molecular mass of approximately 28 kDa, was highly active over an alkaline pH range of 10.0 to 11.0, and remained stable over a pH range of 7.0 to 12.0. The optimum temperature for the enzyme activity was found to be 40~60°C, while the thermotolerance of the enzyme was poor. Therefore, these characteristics of the protease indicate its potential for a wide range of commercial applications.

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