scholarly journals Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications

2022 ◽  
Vol 82 ◽  
Author(s):  
M. K. S. Silva ◽  
T. A. Silva ◽  
J. A. F. Silva ◽  
L. D. A. Costa ◽  
M. L. E. Leal ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Partial Purification ◽  
Biotechnological Applications ◽  
Salting Out ◽  
Process Yield ◽  
Aspartic Proteases ◽  
Biotechnological Potential ◽  
Ph Range ◽  
Pepstatin A ◽  
By Products

Abstract The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by-products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U⋅mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

scholarly journals Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766)

2022 ◽  
Vol 82 ◽  
Author(s):  
J. A. F. Silva ◽  
M. K. S. Silva ◽  
T. A. Silva ◽  
L. D. A. Costa ◽  
M. L. E. Leal ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Partial Purification ◽  
Salting Out ◽  
Aspartic Proteases ◽  
Total Inhibition ◽  
Crevalle Jack ◽  
Pepstatin A ◽  
Caranx Hippos ◽  
Do So

Abstract This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

scholarly journals The Inflammatory Response Induced by Aspartic Proteases of Candida albicans Is Independent of Proteolytic Activity

2010 ◽  
Vol 78 (11) ◽  
pp. 4754-4762 ◽  
Author(s):  
Donatella Pietrella ◽  
Anna Rachini ◽  
Neelam Pandey ◽  
Lydia Schild ◽  
Mihai Netea ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Inflammatory Response ◽  
Proteolytic Activity ◽  
Necrosis Factor Alpha ◽  
Pathogenic Yeast ◽  
Aspartic Proteases ◽  
Akt Activation ◽  
Tnf Α ◽  
Factor Alpha ◽  
Pepstatin A

ABSTRACT The secretion of aspartic proteases (Saps) has long been recognized as a virulence-associated trait of the pathogenic yeast Candida albicans. In this study, we report that different recombinant Saps, including Sap1, Sap2, Sap3, and Sap6, have differing abilities to induce secretion of proinflammatory cytokines by human monocytes. In particular Sap1, Sap2, and Sap6 significantly induced interleukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α), and IL-6 production. Sap3 was able to stimulate the secretion of IL-1β and TNF-α. All Saps tested were able to induce Ca2+ influx in monocytes. Treatment of these Saps with pepstatin A did not have any effect on cytokine secretion, indicating that their stimulatory potential was independent from their proteolytic activity. The capacity of Saps to induce inflammatory cytokine production was also independent from protease-activated receptor (PAR) activation and from the optimal pH for individual Sap activity. The interaction of Saps with monocytes induced Akt activation and phosphorylation of IκBα, which mediates translocation of NF-κB into the nucleus. Overall, these results suggest that individual Sap proteins can induce an inflammatory response and that this phenomenon is independent from the pH of a specific host niche and from Sap enzymatic activity. The inflammatory response is partially dependent on Sap denaturation and is triggered by the Akt/NF-κB activation pathway. Our data suggest a novel, activity-independent aspect of Saps during interactions of C. albicans with the host.

Isolation, Partial Purification and Characterisation of A Plasminogen Activator Produced by Endothelial Cells in Vitro

1979 ◽  
Author(s):  
W.E. Laug
Keyword(s):  
Endothelial Cells ◽  
Proteolytic Activity ◽  
Culture Medium ◽  
Partial Purification ◽  
Diisopropyl Fluorophosphate ◽  
Cell Lysate ◽  
Endothelial Cell Cultures ◽  
Ph Range ◽  
Confluent Endothelial Cell

Cloned endothelial cells obtained from the aorta of 1-2 day old calves produced high fibrinolytic activity, which was 90% dependent upon the presence of plasminogen when grown on 125 I fibrin coated dishes. High plasminogen-dependent proteolytic activity was also demonstrated in the cell lysate and in the culture medium of the cells. The production and secretion of this prtitease were found to increase during the log phase of cell growth and to reach a maximum at con fluency. Thereafter they remained constantly high. This protease, partially purified from the culture medium of confluent endothelial cell cultures, is aiginine specific and activates plasminogen by piOteolytic cleavage to plasmin. Its proteolytic activity which is highest in the pH range of 7.5 to 8.0 is irreversibly inhibited by diisopropyl fluorophosphate, suggesting that it is a serine protease. The molecular weight of this protease is approximately S2000.

scholarly journals Optimal pH in chlorinated swimming pools – balancing formation of by-products

2013 ◽  
Vol 11 (3) ◽  
pp. 465-472 ◽  
Author(s):  
Kamilla M. S. Hansen ◽  
Hans-Jørgen Albrechtsen ◽  
Henrik R. Andersen
Keyword(s):  
Body Fluid ◽  
Product Formation ◽  
Swimming Pools ◽  
Batch Experiments ◽  
Ph Values ◽  
Precursor Ratio ◽  
Decreasing Ph ◽  
Ph Range ◽  
By Products ◽  
Optimal Ph

In order to identify the optimal pH range for chlorinated swimming pools, the formation of trihalomethanes, haloacetonitriles and trichloramine was investigated in the pH-range 6.5–7.5 in batch experiments. An artificial body fluid analogue was used to simulate bather load as the precursor for by-products. The chlorine-to-precursor ratio used in the batch experiments influenced the amounts of by-products formed, but regardless of the ratio the same trends in the effect of pH were observed. Trihalomethane formation was reduced by decreasing pH, but haloacetonitrile and trichloramine formation increased. To evaluate the significance of the increase and decrease of the investigated organic by-products at the different pH values, the genotoxicity was calculated based on literature values. The calculated genotoxicity was approximately at the same level in the pH range 6.8–7.5 and increased when pH was 6.7 or lower. An optimal pH range for by-products formation in swimming pools was identified at pH 7.0–7.2. In the wider pH range (pH 6.8–7.5), the effect on by-product formation was negligible. Swimming pools should never be maintained at lower pH than 6.8 since formation of both haloacetonitriles and trichloramine increase significantly below this value.

scholarly journals DEVELOPMENT AND EVALUATION OF MUCOADHESIVE NANOGEL OF NEVIRAPINE FOR VAGINAL APPLICATION

2019 ◽  
pp. 144-149
Author(s):  
Sheikh Sofiur Rahman ◽  
ABDUL BAQUEE AHMED
Keyword(s):  
Hiv Infection ◽  
Ex Vivo ◽  
Vaginal Fluid ◽  
Content Uniformity ◽  
Vaginal Mucosa ◽  
Salting Out ◽  
Zero Order Kinetics ◽  
The Stability ◽  
Ph Range

Objectives: The main objective of this study was to develop and evaluate Nevirapine nanoparticle loaded mucoadhesive gel (NVP-Np mucoadhesive gel) for vaginal application for the treatment of HIV infection.  Methods: NVP loaded nanoparticles were prepared by salting out method followed by incorporation in different gel bases to produce NVP-Np mucoadhesive gel The prepared gels were evaluated for their physicochemical parameters, rheological characteristics, mucoadhesion, in-vitro drug release and ex-vivo permeation of drug across porcine vaginal mucosa.  Results: The result of FT-IR and DSC study confirmed the absence of incompatibility of NVP with excipients used in the formulations. The particle size of the prepared NVP-Np was found to be 243.8 ± 3.15 nm, a polydispersity index (PI) of 0.787± 0.002 and zeta potential value -17.12 mV, which revealed the stability of nanoparticles. All the formulations showed good homogeneity, spreadability, physical appearance and content uniformity. The pH of the mucoadhesive gel formulations was in the range of 3.70 ± 0.03 to 4.56 ± 0.02, which lies in the normal pH range of the vaginal fluid.  The cumulative amounts permeated at 6 h were 832.23 ± 63.45 μg/cm2 , 592.13 ± 82.55 μg/cm2 and 941.32 ± 81.10 μg/cm2 from F1(1% Chitosan), F2(1% Carbopol 974P) and F3 (1% HPMC K100M )  respectively. A linear relationship [r2 > 0.9 (0.97 n 0.99)] was observed between the percentage cumulative amount permeated and time, indicating zero order kinetics. Conclusion: In conclusion, NVP-Np mucoadhesive gel was prepared successfully using salting out followed by a homogenization technique for vaginal application of NVP for the prophylaxis of HIV infection.

Genomics Perspectives on Metabolism, Survival Strategies, and Biotechnological Applications of Brettanomyces bruxellensis LAMAP2480

2017 ◽  
Vol 27 (3) ◽  
pp. 147-158 ◽  
Author(s):  
Liliana Godoy ◽  
Evelyn Silva-Moreno ◽  
Wladimir Mardones ◽  
Darwin Guzman ◽  
Francisco A. Cubillos ◽  
...  
Keyword(s):  
Ethanol Production ◽  
Survival Strategies ◽  
Whole Genome ◽  
Biotechnological Applications ◽  
Genome Sequences ◽  
Wine Production ◽  
Brettanomyces Bruxellensis ◽  
Biotechnological Potential ◽  
Spoilage Yeast ◽  
Genome Comparisons

Wine production is an important commercial issue for the liquor industry. The global production was estimated at 275.7 million hectoliters in 2015. The loss of wine production due to <i>Brettanomyces bruxellensis </i>contamination is currently a problem. This yeast causes a “horse sweat” flavor in wine, which is an undesired organoleptic attribute. To date, 6 <i>B. bruxellensis </i>annotated genome sequences are available (LAMAP2480, AWRI1499, AWRI1608, AWRI1613, ST05.12/22, and CBS2499), and whole genome comparisons between strains are limited. In this article, we reassembled and reannotated the genome of <i>B. bruxellensis</i> LAMAP2480, obtaining a 27-Mb assembly with 5.5 kb of N50. In addition, the genome of <i>B. bruxellensis</i> LAMAP2480 was analyzed in the context of spoilage yeast and potential as a biotechnological tool. In addition, we carried out an exploratory transcriptomic analysis of this strain grown in synthetic wine. Several genes related to stress tolerance, micronutrient acquisition, ethanol production, and lignocellulose assimilation were found. In conclusion, the analysis of the genome of <i>B. bruxellensis</i> LAMAP2480 reaffirms the biotechnological potential of this strain. This research represents an interesting platform for the study of the spoilage yeast <i>B. bruxellensis</i>.

Isolation and partial purification of phytotoxins from liquid cultures of Leucostoma cincta and Leucostoma persoonii

1991 ◽  
Vol 69 (9) ◽  
pp. 1998-2003 ◽  
Author(s):  
A. M. Svircev ◽  
A. R. Biggs ◽  
N. W. Miles
Keyword(s):  
Isoelectric Focusing ◽  
Prunus Persica ◽  
Partial Purification ◽  
Liquid Cultures ◽  
Phytotoxic Activity ◽  
Stem Necrosis ◽  
Cold Acetone ◽  
Characterization Studies ◽  
Ph Range ◽  
Cytospora Canker

Phytotoxins from Leucostoma persoonii and Leucostoma cincta were isolated by sequential ultrafiltration of cell-free culture filtrates through a series of exclusion membranes. Phytotoxic activity, characterized by stem necrosis and leaf wilt, chlorosis, and necrosis, was evident when a cold acetone precipitate from the < 1000-Da fraction was tested in a peach shoot tip bioassay. Analytical polyacrylamide isoelectrofocusing of the < 1000-Da fraction in the pH range 3.5–6.0 identified two peptide bands at pH 4.0 and 6.0. Preparative granulated bed isoelectric focusing in the same pH range yielded two fractions with phytotoxic activity against peach shoots. Preliminary characterization studies indicate that the two fractions with toxin activity contain small polypeptides. Key words: Cytospora spp., cytospora canker, Prunus persica, peach.

scholarly journals Screening, Identification of Alkaline Proteases Producing Fungi from Soil of Different Habitats of Amalner Tahsil [Maharashtra] and Their Applications

2017 ◽  
Vol 5 (3) ◽  
pp. 397-402 ◽  
Author(s):  
H.K. Suryawanshi ◽  
N.D. Pandya
Keyword(s):  
Proteolytic Activity ◽  
Soil Samples ◽  
Precipitation Method ◽  
Diffusion Method ◽  
Clot Lysis ◽  
Partial Purification ◽  
Fungal Isolation ◽  
Lysis Activity ◽  
Stain Removal ◽  
Pharmaceutical Industries

Fungal proteases had wide applications in textile, leather, food and Pharmaceutical industries. As proteases shows proteolytic activity they are helpful in proteinic stain removal hence also used in various commercial detergent industries. For fungal isolation soil samples were collected from different sites of Amalner tahsil. (Maharashtra) e.g. crop fields, near dairy areas, poultry farms etc. Those soil samples showing alkaline pH were selected for isolation of fungi on Potato Dextrose Agar plates. Then among 14 different isolates 2 were selected for their most proteolytic activity after screening on casein agar, skimmed milk agar and gelatine agar. For submerged fermentation, these selected isolates were inoculated in production media in shake flask. After 72 hrs, plate assay was performed by taking crude enzyme after filtration and centrifugation as well as by taking partially purified enzyme.(partial purification done by ammonium sulphate precipitation method). Protease activity assay was performed by agar well diffusion method, as well as blood clot lysis activity and blood stain removal ability of protease obtained from selected isolates was studied.Selected isolates were identified,among them Aspergillus niger gives more proteolytic activity than Trichoderma longibrachiatum. Int. J. Appl. Sci. Biotechnol. Vol 5(3): 397-402

scholarly journals Conversion of Shrimp Head Waste for Production of a Thermotolerant, Detergent-Stable, Alkaline Protease by Paenibacillus sp.

Catalysts ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 798 ◽  
Author(s):  
Chien Thang Doan ◽  
Thi Ngoc Tran ◽  
I-Hong Wen ◽  
Van Bon Nguyen ◽  
Anh Dzung Nguyen ◽  
...  
Keyword(s):  
Alkaline Protease ◽  
Nitrogen Nutrition ◽  
Crab Shell ◽  
Carbon And Nitrogen ◽  
Optimal Activity ◽  
Shrimp Shell ◽  
Paenibacillus Sp ◽  
Shrimp Head ◽  
Ph Range ◽  
By Products

Fishery processing by-products have been of great interest to researchers due to their beneficial applications in many fields. In this study, five types of marine by-products, including demineralized crab shell, demineralized shrimp shell, shrimp head, shrimp shell, and squid pen, provided sources of carbon and nitrogen nutrition by producing a protease from Paenibacillus sp. TKU047. Strain TKU047 demonstrated the highest protease productivity (2.98 U/mL) when cultured for two days on a medium containing 0.5% of shrimp head powder (SHP). The mass of TKU047 protease was determined to be 32 kDa (approximately). TKU047 protease displayed optimal activity at 70–80 °C and pH 9, with a pH range of stability from 6 to 11. TKU047 protease also showed stability in solutions containing surfactants and detergents. Based on its excellent properties, Paenibacillus sp. TKU047 protease may be a feasible candidate for inclusion in laundry detergents.

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