scholarly journals Bacillus sp. CSK2 produced thermostable alkaline keratinase using agro-wastes: keratinolytic enzyme characterization

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Nonso E. Nnolim ◽  
Uchechukwu U. Nwodo
Keyword(s):  
Incubation Temperature ◽  
Ethylenediaminetetraacetic Acid ◽  
Proteolytic Enzymes ◽  
Catalytic Efficiency ◽  
Tween 80 ◽  
Enzyme Characterization ◽  
Chicken Feathers ◽  
Laundry Detergents ◽  
Poultry Processing ◽  
Optimized Conditions

Abstract Background Chicken feathers are the most abundant agro-wastes emanating from the poultry processing farms and present major concerns to environmentalists. Bioutilization of intractable feather wastes for the production of critical proteolytic enzymes is highly attractive from both ecological and biotechnological perspectives. Consequently, physicochemical conditions influencing keratinase production by Bacillus sp. CSK2 on chicken feathers formulation was optimized, and the keratinase was characterized. Results The highest enzyme activity of 1539.09 ± 68.14 U/mL was obtained after 48 h of incubation with optimized conditions consisting of chicken feathers (7.5 g/L), maltose (2.0 g/L), initial fermentation pH (5.0), incubation temperature (30 °C), and agitation speed (200 rpm). The keratinase showed optimal catalytic efficiency at pH 8.0 and a temperature range of 60 °C – 80 °C. The keratinase thermostability was remarkable with a half-life of above 120 min at 70 °C. Keratinase catalytic efficiency was halted by ethylenediaminetetraacetic acid and 1,10-phenanthroline. However, keratinase activity was enhanced by 2-mercaptoethanol, dimethyl sulfoxide, tween-80, but was strongly inhibited by Al3+ and Fe3+. Upon treatment with laundry detergents, the following keratinase residual activities were achieved: 85.19 ± 1.33% (Sunlight), 90.33 ± 5.95% (Surf), 80.16 ± 2.99% (Omo), 99.49 ± 3.11% (Ariel), and 87.19 ± 0.26% (Maq). Conclusion The remarkable stability of the keratinase with an admixture of organic solvents or laundry detergents portends the industrial and biotechnological significance of the biocatalyst.

scholarly journals Enhanced Production, Purification and Characterization of Alkaline Keratinase from Streptomyces minutiscleroticus DNA38

2015 ◽  
Vol 43 ◽  
pp. 27-37 ◽  
Author(s):  
Allure Nandini ◽  
D.N. Madhusudhan ◽  
Agsar Dayanand
Keyword(s):  
Yeast Extract ◽  
Incubation Temperature ◽  
Direct Action ◽  
Interactive Effect ◽  
Chicken Feathers ◽  
Enhanced Production ◽  
Optimized Conditions ◽  
Critical Process ◽  
Central Composite

A thermo tolerant, feather-degrading, newly isolated actinobacterial strain Streptomyces minutiscleroticus DNA38 was investigated for its ability to produce keratinase. Maximum production (283.4 IU) of keratinase by Streptomyces minutiscleroticus DNA38 in starch chicken feathers medium under submerged bioprocess was observed at optimized conditions of pH 9.0 of the medium and 45 °C incubation temperature. Further, an enhanced production (435.8 IU) of keratinase was achieved employing response surface methodology. Combined interactive effect of starch (7.50 g/L), yeast extract (0.74 g/L) and chicken feathers (7.50 g/L) were found to be the critical process variables for enhanced production under central composite design. Chicken feathers showed a direct action and addition of starch and yeast extract to the medium proved effective for a significant increase in the production of keratinase. The purified keratinase was monomeric and had a molecular mass of 29 kDa. The enzyme activity was significantly inhibited after pH 9.0 and temperature 50 °C.

scholarly journals Molecular Elucidation of a Urate Oxidase from Deinococcus radiodurans for Hyperuricemia and Gout Therapy

2021 ◽  
Vol 22 (11) ◽  
pp. 5611
Author(s):  
Yi-Chih Chiu ◽  
Ting-Syuan Hsu ◽  
Chen-Yu Huang ◽  
Chun-Hua Hsu
Keyword(s):  
Uric Acid ◽  
High Stability ◽  
Deinococcus Radiodurans ◽  
Catalytic Efficiency ◽  
Enzyme Characterization ◽  
Urate Oxidase ◽  
Medical Applications ◽  
Physiological Conditions ◽  
Extremophile Microorganism ◽  
Mesophilic Conditions

Urate oxidase initiates the uric acid degradation pathways and is extensively used for protein drug development for gout therapy and serum uric acid diagnosis. We first present the biochemical and structural elucidation of a urate oxidase from the extremophile microorganism Deinococcus radiodurans (DrUox). From enzyme characterization, DrUox showed optimal catalytic ability at 30 °C and pH 9.0 with high stability under physiological conditions. Only the Mg2+ ion moderately elevated its activity, which indicates the characteristic of the cofactor-free urate oxidase family. Of note, DrUox is thermostable in mesophilic conditions. It retains almost 100% activity when incubated at 25 °C and 37 °C for 24 h. In this study, we characterized a thermostable urate oxidase, DrUox with high catalytic efficiency and thermal stability, which strengthens its potential for medical applications.

scholarly journals Artificial Neural Networks and Response Surface Methodology Approach for Optimization of an Eco-Friendly and Detergent-Stable Lipase Production from Actinomadura Keratinilytica Strain Cpt29

2021 ◽  
Vol 68 (3) ◽  
pp. 575-586
Author(s):  
Noura Semache ◽  
Fatiha Benamia ◽  
Bilal Kerouaz ◽  
Inès Belhaj ◽  
Selma Bounour ◽  
...  
Keyword(s):  
Incubation Temperature ◽  
Tween 80 ◽  
Fold Increase ◽  
Data Fitting ◽  
Lipase Production ◽  
Potential Candidate ◽  
Cultivation Time ◽  
North East ◽  
Lipase A ◽  
Commercial Laundry

This work mainly focused on the production of an efficient, economical, and eco-friendly lipase (AKL29) from Actinomadura keratinilytica strain Cpt29 isolated from poultry compost in north east of Algeria, for use in detergent industries. AKL29 shows a significant lipase activity (45 U/mL) towards hydrolyzed triacylglycerols, indicating that it is a true lipase. For maximum lipase production the modeling and optimization of potential culture parameters such as incubation temperature, cultivation time, and Tween 80 (v/v) were built using RSM and ANN approaches. The results show that both the two models provided good quality predictions, yet the ANN showed a clear superiority over RSM for both data fitting and estimation capabilities. A 4.1-fold increase in lipase production was recorded under the following optimal condition: incubation temperature (37.9 °C), cultivation time (111 h), and Tween 80 (3.27%, v/v). Furthermore, the partially purified lipase showed good stability, high compatibility, and significant wash performance with various commercial laundry detergents, making this novel lipase a promising potential candidate for detergent industries.

scholarly journals Extruction of keratinase from the local isolate Bacillus licheniformis and partially purified and characterized

2009 ◽  
Vol 3 (2) ◽  
pp. 41-52
Author(s):  
Rasha T. Abdullah ◽  
Abdulkareem J. Hashim ◽  
JASIM M. Karhout
Keyword(s):  
Enzyme Activity ◽  
Bovine Serum Albumin ◽  
Ion Exchange ◽  
Bacillus Licheniformis ◽  
Enzyme Characterization ◽  
Optimal Temperature ◽  
Chicken Feathers ◽  
Local Isolate ◽  
Cellulose Column ◽  
Optimal Ph

The keratinase produced from local isolate Bacillus licheniformis was purified by two steps included precipitation by ammonium sulphate with 40% saturation; followed by ion exchange using CM-Cellulose column. The enzyme was purified to 12.6 times in the last step with an enzyme yield of 17%. Enzyme characterization results indicated that: The optimal pH for enzyme activity was 7.5 and it was stable at 7-9.5. The optimal temperature for enzyme activity was 50°C and it was stable for 30 min at 25-45 °C. Substrate specifity was tested using casein, Bovine serum albumin, gelatin, hooves, human hair, chicken feathers and wool; higher specifity was recorded using casein gave 0.6 unit /ml. The enzyme was inhibited by PMSF and metal ions like Hg+2, Fe+2, Cu+2 and Mn+2, and activated by Ca+2, Mg+2, Zn+2and Al+3.

scholarly journals Canola Cake as a Potential Substrate for Proteolytic Enzymes Production by a Selected Strain of Aspergillus oryzae: Selection of Process Conditions and Product Characterization

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Adriana C. Freitas ◽  
Ruann J. S. Castro ◽  
Maria A. Fontenele ◽  
Antonio S. Egito ◽  
Cristiane S. Farinas ◽  
...  
Keyword(s):  
Aspergillus Oryzae ◽  
Protease Activity ◽  
Incubation Temperature ◽  
Initial Conditions ◽  
Proteolytic Enzymes ◽  
Inoculum Size ◽  
Protease Production ◽  
Process Conditions ◽  
Enzymatic Extract ◽  
Technological Advances

Oil cakes have excellent nutritional value and offer considerable potential for use in biotechnological processes that employ solid-state fermentation (SSF) for the production of high value products. This work evaluates the feasibility of using canola cake as a substrate for protease production by a selected strain of Aspergillus oryzae cultivated under SSF. The influences of the following process parameters were considered: initial substrate moisture content, incubation temperature, inoculum size, and pH of the buffer used for protease extraction and activity analysis. Maximum protease activity was obtained after cultivating Aspergillus oryzae CCBP 001 at 20°C, using an inoculum size of 107 spores/g in canola cake medium moistened with 40 mL of water to 100 g of cake. Cultivation and extraction under selected conditions increased protease activity 5.8-fold, compared to the initial conditions. Zymogram analysis of the enzymatic extract showed that the protease molecular weights varied between 31 and 200 kDa. The concentrated protease extract induced clotting of casein in 5 min. The results demonstrate the potential application of canola cake for protease production under SSF and contribute to the technological advances needed to increase the efficiency of processes designed to add value to agroindustrial wastes.

scholarly journals Isolation of Bacillus Strains with Keratinolitic Activity

2020 ◽  
Author(s):  
S. Aktayeva ◽  
K. Baltin ◽  
B. Khassenov
Keyword(s):  
Proteolytic Enzymes ◽  
Sole Source ◽  
Environmental Safety ◽  
Economic Feasibility ◽  
Bacillus Sp ◽  
Optimal Conditions ◽  
Processing Waste ◽  
Keratinolytic Activity ◽  
Chicken Feathers ◽  
Promising Source

Environmental safety and economic feasibility determine the search for new ways of processing waste in poultry farms. Most of this waste is down and feathers, which are 90% β-keratin. Feathers can be a valuable source of amino acids and peptones when properly processed. The most effective is enzymatic treatment of feather keratin. The search for new strains producing keratinolytic enzymes seems to be a promising direction. On the territory of the poultry farm, 4 strains were isolated from the places of accumulation of feathers. They are able to use chicken feathers as their sole source of organic matter. Based on morphological, genomic, and proteomic analyzes, the isolated strains were identified as Bacillus sp. It was found that the strains secrete proteolytic enzymes that hydrolyze collagen, casein, β-keratin and do not hydrolyze bovine serum albumin. Feather hydrolysis experiments showed that the Bacillus sp. A5.3 possesses maximum keratinolytic activity, and on the second day, the destruction of the second order barbs is observed. The keratinase activity of the strain on azokeratin after an hour of incubation on feather medium was 27.4 U/ml. The optimal conditions for the complex of secreted proteolytic enzymes are pH 7.0-8.0 and temperature 35-40 °C. The isolated Bacillus sp. A5.3 strain is a promising source of proteases and keratinases.

Sensitivity to proteolytic enzymes in laundry detergents

1971 ◽  
Vol 47 (2) ◽  
pp. 76-79
Author(s):  
Richard S. Shapiro ◽  
B.S. Pharm ◽  
Ben C. Eisenberg
Keyword(s):  
Proteolytic Enzymes ◽  
Laundry Detergents

Uptake of latex particles by macrophages: characterization using flow cytometry

1983 ◽  
Vol 245 (3) ◽  
pp. C220-C226 ◽  
Author(s):  
R. J. Parod ◽  
J. D. Brain
Keyword(s):  
Flow Cytometry ◽  
Incubation Temperature ◽  
Ethylenediaminetetraacetic Acid ◽  
Divalent Cations ◽  
Small Samples ◽  
Fluorescent Intensity ◽  
Latex Particles ◽  
Particle Uptake ◽  
Pulmonary Macrophages ◽  
Cell To Cell Variability

Flow cytometry can be used to characterize the uptake of particles by small samples of pulmonary macrophages both quickly and accurately. We found that there was a linear relationship between the number of fluorescent latex particles and the fluorescent intensity associated with each cell up to 47 particles/cell. Macrophages lavaged from the lungs of Syrian golden hamsters were metabolically and functionally stable for 3 h of incubation; the average intracellular concentrations of Na+ and K+ were 29 +/- 2 and 158 +/- 6 mM, respectively. Particle uptake was significantly inhibited by removing divalent cations with ethylenediaminetetraacetic acid and lowering the incubation temperature (37 degrees C) to 24 and 3 degrees C. The cell-to-cell variability in the uptake of particles was greater than the Poisson distribution would predict based on the mean number of particles per cell.

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.

Enzyme-linked Immunosorbent Assay for a Soluble Antigen of Renibacterium salmoninarum, the Causative Agent of Salmonid Bacterial Kidney Disease

1987 ◽  
Vol 44 (1) ◽  
pp. 183-191 ◽  
Author(s):  
R. J. Pascho ◽  
D. Mulcahy
Keyword(s):  
Immunosorbent Assay ◽  
Incubation Temperature ◽  
Tween 80 ◽  
Broth Culture ◽  
Tween 20 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Soluble Antigen ◽  
Renibacterium Salmoninarum ◽  
Bacterial Kidney Disease ◽  
Wash Buffer

A double-antibody enzyme-linked immunosorbent assay (ELISA) for detection of a soluble fraction of Renibacterium salmoninarum was developed from components extracted from the supernatant of an R. salmoninarum broth culture. The Costar® Serocluster™ EIA microplate gave the highest absorbance and signal-to-noise ratios among seven types tested. Including Tween 80 in the wash buffer resulted in higher absorbances than Tween 20 when antigen was present. Background absorbance did not increase when Tween 80 was added to the wash buffer, but did when Tween 80 replaced Tween 20 in antigen and conjugate diluents. Adsorption of coating antibody peaked within 4 h at 37 °C and 16 h at 4 °C. Antigen attachment to antibody-coated microplate wells depended more on incubation temperature than duration; we adopted a 3-h incubation at 25 °C. Conjugate incubation for longer than 1 h at 37 °C or 3 h at 25 °C resulted in unacceptable background levels. No cross-reactions resulted from heat-extracted antigens of 10 other species of bacteria. The optimized ELISA is a 6-h test that enables detection of levels of soluble antigen as low as 2–20 ng.

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