Some properties of the extracellular proteolytic enzymes of the milk-spoiling organismPseudomonas aeruginosaATCC 10145

1968 ◽  
Vol 35 (3) ◽  
pp. 395-398 ◽  
Author(s):  
H. S. Juffs ◽  
H. W. Doelle
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Proteolytic Activity ◽  
Culture Supernatant ◽  
Proteolytic Enzymes ◽  
Maximum Activity ◽  
Ph Range

SummaryThe proteolytic enzymes present in the culture supernatant ofPseudomonas aeruginosaATCC 10145 were active in the pH range 5·5–9·0 with a maximum activity at pH 7·3. Heating for 15 sec at 72°C resulted in a 36% loss of proteolytic activity whereas heating for 30 min at 63°C resulted, in a 6% loss. Boiling for 2 min completely inactivated the proteolytic enzymes. At 2°C the proteolytic enzymes were stable for at least a month and casein was readily hydrolysed at this temperature.

Growth and proteinase production inPseudomonasspp. cultivated under various conditions of temperature and nutrition

1968 ◽  
Vol 35 (3) ◽  
pp. 385-393 ◽  
Author(s):  
H. S. Juffs ◽  
A. C. Hayward ◽  
H. W. Doelle
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Proteolytic Activity ◽  
Proteolytic Enzyme ◽  
Organic Nitrogen ◽  
Proteolytic Enzymes ◽  
Dry Weight ◽  
Growth Cycle ◽  
Logarithmic Phase ◽  
Proteinase Production

SummaryA study was made of the formation of the extracellular proteolytic enzymes during the growth cycle of several species ofPseudomonascultivated under different conditions of temperature and nutrition. Proteolytic activity was not proportional to growth. Expressed per unit of cell dry weight, the proteolytic activity showed a peak in the early logarithmic phase which was greater in cultures grown at 3 than at 28°C. Proteolytic enzyme was not formed in the absence of organic nitrogen. Of 16 organisms studied,Pseudomonas aeruginosaATCC 10145 was the most prolific producer of proteolytic enzyme.

Enzymic Proteolysis byEntamoeba histolytica; biochemical characteristics and relationship with invasiveness

Parasitology ◽  
1960 ◽  
Vol 50 (3-4) ◽  
pp. 531-550 ◽  
Author(s):  
R. A. Neal
Keyword(s):  
Proteolytic Activity ◽  
Culture Media ◽  
Proteolytic Enzymes ◽  
Egg Yolk ◽  
Single Strain ◽  
High Proteolytic Activity ◽  
Serum Inhibitor ◽  
Sulphydryl Groups ◽  
Ph Range

The proteolytic activity of extracts ofEntamoeba histolyticahas been further investigated. Casein and gelatin, but not haemaglobin, were hydrolysed. Activity was observed in the pH range 5·8 to 8·5 with optimal activity at pH 7·5 to 7·9. Activity was optimal at 37° C. and sulphydryl groups were not required. Concomitant bacteria showed no proteolytic activity. Hyaluronidase, collagenase and lecithinase could not be detected.An inhibitor of proteolytic enzyme was present in sera of all animals tested and in egg yolk. All culture media prepared from eggs were inhibitory, but inspissation or dilution of serum inactivated the serum inhibitor. Purified trypsin inhibitors from lima and soy bean were not active against the amoebic enzyme. Proteolytic enzymes were not secreted extracellularlyin vitro.High proteolytic activity was found in two out of five invasive, freshly isolated, strains ofE. histolyticaand after two series of liver passages of a single strain. The significance of these observations is discussed. It is concluded that the present evidence does not convincingly demonstrate that high proteolytic activity is required for tissue invasion by amoebae, but may accompany another factor.

Aquaculture by-product: a source of proteolytic enzymes for detergent additives

2009 ◽  
Vol 63 (6) ◽  
Author(s):  
Chirleanny Mendes ◽  
Marília Brito ◽  
Tatiana Porto ◽  
Ana Porto ◽  
Ranilson Bezerra ◽  
...  
Keyword(s):  
Proteolytic Enzymes ◽  
Maximum Activity ◽  
Alkaline Ph ◽  
Detergent Additive ◽  
Sodium Dodecylsulphate ◽  
Detergent Concentration ◽  
Independent Variables ◽  
Nile Tilapia Oreochromis Niloticus ◽  
Ph Range ◽  
Full Factorial

AbstractIntestine proteases of Nile tilapia (Oreochromis niloticus) were partially purified by heat treatment (purification factor of 3.5, enzyme activity remained almost constant) to reach the maximum activity and stability within an alkaline pH range of 7.2–11.0. The optimum temperature and stability over a 120 min period were found to be at 55°C and at 35–45°C, respectively. The proteases’ activity was not affected by a 1 vol. % saponin surfactant, inactivated by 0.01 g mL−1 sodium dodecylsulphate after 120 min, and it remained stable for 30 min in a 5 vol. % and 10 vol. % hydrogen peroxide solutions. The proteases were slightly activated by Ca2+, Mg2+, and K+ and the substrate most effectively hydrolysed was casein (40.0 U mg−1). A 24 full factorial design used to evaluated the influence of independent variables showed that the enzyme extract, detergent concentration and the incubation time had a significant influence on the enzymatic activity. The best conditions to be used concerning detergent additive were found with 0.3 mg mL−1 of protein and 3.0 mg mL−1 of detergent for 30 min in the presence of Astrus® detergent.

An initial characterization of the proteolytic enzymes secreted by the adult stage of the human hookworm Necator americanus

Parasitology ◽  
1995 ◽  
Vol 110 (5) ◽  
pp. 555-563 ◽  
Author(s):  
A. Brown ◽  
J. M. Burleigh ◽  
E. E. Billett ◽  
D. I. Pritchard
Keyword(s):  
Serine Proteases ◽  
Metal Ion ◽  
Proteolytic Enzymes ◽  
Maximum Activity ◽  
Biologically Relevant ◽  
Necator Americanus ◽  
Recent Developments ◽  
Proteolytic Activities ◽  
Secretory Products ◽  
Ph Range

SUMMARYThe proteolytic activities present in adult Necator americanus excretory–secretory products have been assessed using biologically relevant, naturally occurring substrates (haemoglobin and fibrinogen) and a number of synthetic fluorogenic and chromogenic substrates. One broad peak of activity was observed against haemoglobin in the pH range 5 to 7, with maximum activity at pH 6·6, while fibrinogenolytic activity was shown to be greater at pH 3·5. Inhibition studies against haemoglobin, fibrinogen and synthetic substrates using a battery of appropriate protease inhibitors indicated the presence of a mixture of aspartyl, cysteinyl and serine proteases. Metal ion (Ca2+, Zn2+ and Fe2+) stimulation was demonstrated, with stimulation by Zn2+ being the most marked. These results are discussed in the context of recent developments in the field of parasite proteolytic enzymes, where they have been suggested as targets for immuno- and chemotherapy.

scholarly journals Actividad enzimática de proteasas de Cyprinus carpio (Cypriniformes: Cyprinidae) extraídas de una laguna contaminada en México

2017 ◽  
Vol 65 (2) ◽  
Author(s):  
Arisaí Hernández-Sámano ◽  
Xochitl Guzmán-García ◽  
Raquel García-Barrientos ◽  
Isabel Guerrero-Legarreta
Keyword(s):  
Proteolytic Activity ◽  
Cyprinus Carpio ◽  
Proteolytic Enzymes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Waste Water Treatment ◽  
Sodium Dodecyl ◽  
Maximum Activity ◽  
Protease Production ◽  
Human Consumption ◽  
Alkaline Ph

Common carp (Cyprinus carpio) is an aquatic organism of commercial value able to survive in polluted environments; carps contain proteolytic enzymes of physiological importance and potential industrial application. The objective of this work was partially purify and study the proteolytic activity at different pH of carp proteases living in a polluted environment. Three carps were captured in different zones of Zumpango polluted lagoon (Mexico) at 1 m of maximum deep. Protease crude extracts were obtained from dorsal muscle by aqueous extraction and fractionated by 20 %, 50 %, 80 %-saturated (NH4)2SO4. Fractions extracted with 50 % and 80 %-saturated (NH4)2SO4 were selected for their high proteolytic activity and concentrated by ultrafiltration through 100 kDa molecular weight cutoff membranes and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The crude proteolytic extract had significantly higher activity (19.7 - 20.3 U / mg) at pH 2, 5, and 7 (P < 0.001). Fractions obtained with 20 %, 50 % and 80 % - saturated (NH4)2SO4 showed peak activity at pH 5 (2.8 U / mg) and pH 6 (2.2 U / mg); pH 6 (4.3 U / mg) and pH 3 - 4 (3.6 - 3.7 U / mg); pH 3 (10.8 U / mg) and pH 10 (10.6 U / mg); respectively. Subfractions of < 100 kDa, obtained with 50 % and 80 %-saturated (NH4)2SO4, had peak proteolytic activity at alkaline pH. A < 100 kDa fraction, obtained with 80 %-saturated (NH4)2SO4, had the highest proteolytic activity (37.3 - 43.7 U / mg) at pH 8 - 10, purification factor of 3 and 19.1 % recovery. Thirteen proteins between 9.8 to 104.8 kDa were identified in the crude extract. Peak protein concentration was observed for 31 - 33 and 39 - 41 kDa, suggesting the possibility predominance of serine- and aspartyl- proteases, respectively. We suggest this protease with maximum activity at alkaline pH is related to the adaptation of C. carpio to polluted waters with high pH. Although unsuitable for human consumption, these organisms can be a source of protease production aimed to several uses as in the industry and waste water treatment among others.

scholarly journals Bioprocess development for enhanced endoglucanase production by newly isolated bacteria, purification, characterization and in-vitro efficacy as anti-biofilm of Pseudomonas aeruginosa

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Atef M. Ibrahim ◽  
Ragaa A. Hamouda ◽  
Noura El-Ahmady El-Naggar ◽  
Fatma M. Al-Shakankery
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Yeast Extract ◽  
Statistical Design ◽  
Life Time ◽  
Maximum Activity ◽  
Bioprocess Development ◽  
Ammonium Sulfate Precipitation ◽  
Face Centered ◽  
Central Composite

AbstractEndoglucanase producing bacteria were isolated from Egyptian soils and the most active bacterial strain was identified as Bacillus subtilis strain Fatma/1. Plackett–Burman statistical design was carried out to assess the effect of seven process variables on endoglucanase production. Carboxymethyl cellulose (CMC), yeast extract and peptone were the most significant variables that enhanced the endoglucanase production and thus were selected for further optimization using face-centered central composite design. The highest yield of endoglucanase (32.37 U/mL) was obtained in run no. 9, using 18 g/L CMC, 8 g/L peptone, 7 g/L yeast extract and 0.1 g/L FeSO4.7H2O. The optimized medium showed about eightfold increase in endoglucanase production compared to the unoptimized medium. The produced crude enzyme was further purified by ammonium sulfate precipitation, then DEAE-Sepharose CL6B column. The purified enzyme was shown to have a molecular weight of 37 kDa. The enzyme showed maximum activity at pH 8.0, temperature of 50 °C, incubation time of 60 min. The half-life time (T1/2) was 139.53 min at 50 °C, while being 82.67 min at 60 °C. Endoglucanase at concentration of 12 U/mL effectively removed 84.61% of biofilm matrix of Pseudomonas aeruginosa with marked reduction in carbohydrate content of the biofilm from 63.4 to 7.9 μg.

scholarly journals Comprehensive analysis of the isozyme composition of laccase derived from Japanese lacquer tree, Toxicodendron vernicifluum

2021 ◽  
Vol 67 (1) ◽  
Author(s):  
Mariko Takano ◽  
Masaya Nakamura ◽  
Masanobu Tabata
Keyword(s):  
Isoelectric Focusing ◽  
Laccase Activity ◽  
Maximum Activity ◽  
Acetone Powder ◽  
Blue Color ◽  
Buffer Solution ◽  
Activity Staining ◽  
Water Extracts ◽  
Ph Range ◽  
Toxicodendron Vernicifluum

AbstractWe performed an analysis using isoelectric focusing to comprehensively clarify the isozyme composition of laccase derived from Japanese lacquer tree, Toxicodendron vernicifluum. When water extracts of acetone powder obtained from lacquer were subjected to isoelectric focusing, five bands within pI 7.35–9.30 and nine bands within pI 3.50–5.25 were detected using Coomassie staining. Similarly, laccase activity staining using guaiacol showed five bands within pI 7.35–9.30 and three bands within pI 3.50–4.25. However, laccase activity staining using gallic acid showed remarkable staining within pI 3.50–5.85, whereas staining was very weak within pI 7.35–9.30. When the water extracts of acetone powder were fractionated into the fractions containing bands within pI 7.35–9.30 and pI 3.50–5.85 by SP-Sepharose column chromatography, the former had a blue color and the latter a yellow color. The laccase activity was measured for each of the fractions in buffer solution in the pH range of 2.5–8.0. When syringaldazine, guaiacol, and 2,6-dimethoxyphenol were used as substrates, the yellow fraction showed considerably higher activity than the blue fraction for pH 5.5–7.5. When 3-methylcatechol and 4-methylcatechol were used as substrates, the yellow fraction showed higher activity for pH 4.5–6.5, and the blue fraction showed higher activity for pH 7.0–8.0. When 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) was used as the substrate, both fractions showed maximum activity at optimum pH of 3.0–4.0. Conventionally, in research on blue laccase derived from lacquer, the non-blue fraction corresponding to the yellow fraction lower than pI 6 has been removed during the purification process and thus has not been analyzed. Our results indicated that yellow laccase was present in the non-blue components of lacquer and that it may play a role in urushiol polymerization with previously reported blue laccase.

Pyridine–Adenine Dinucleotide Transhydrogenase Activity in Cells Cultured from Rat Hepatoma

1972 ◽  
Vol 50 (5) ◽  
pp. 447-456 ◽  
Author(s):  
C. De Luca ◽  
R. P. Gioeli
Keyword(s):  
Phosphate Buffer ◽  
Pyridine Nucleotide ◽  
Maximum Activity ◽  
Ph Optimum ◽  
Apparent Lack ◽  
Minimal Deviation ◽  
Ph Range ◽  
Pyridine Nucleotide Transhydrogenase ◽  
Rat Hepatoma ◽  
Cells Cultured

Preparations from cells cultured from a minimal-deviation hepatoma in the rat exhibit pyridine nucleotide transhydrogenase (NAD(P)H: NAD(P) oxidoreductase, EC 1.6.1.1) activity. The pH optimum, its release by digitonin, and its apparent lack of dependence on steroids for activity tentatively classify it as a transhydrogenase of the type first described for animal tissue.Enzyme preparations from digitonin-treated homogenates were very unstable. The time necessary for the loss of one-half the activity was 16–18 h when the enzyme was stored at 5 °C; this was reduced to 4 h when storage was in polycarbonate tubes.The enzyme apparently transferred hydrogen directly and with equal ease from NADH to both the 3-acetyl-pyridine and thionicotinamide analogues of NAD. Half-saturation values for NAD and its acetylpyridine analogue were 0.99 × 10−5 M and 3.55 × 10−4 M, respectively. The enzyme exhibited its maximum activity in phosphate buffer at pH 5.8. It was inhibited by 50–60% over the pH range 7.0–8.5 in Tris buffer. This could be reversed by dithiothreitol; reversal was complete between pH 8.0 and 8.5.

Growth of Salmonella Enteritidis in the presence of quorum sensing signaling compounds produced by Pseudomonas aeruginosa

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Weijia He ◽  
Huamei Yang ◽  
Xiang Wang ◽  
Hongmei Li ◽  
Qingli Dong
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Salmonella Enteritidis ◽  
Culture Supernatant ◽  
Lag Time ◽  
Bacterial Communication ◽  
Homoserine Lactones ◽  
Positive Effects ◽  
Kinetics Of

Abstract Quorum sensing (QS) can exist in food-related bacteria and potentially affect bacterial growth through acyl-homoserine lactones (AHLs). To verify the role of QS compounds in the cell-free supernatant, this study examined the effect of supernatant extracted from Pseudomonas aeruginosa culture on the growth kinetics of Salmonella Enteritidis. The results showed that the lag time (λ) of S. Enteritidis was apparently reduced (p < 0.05) under the influence of P. aeruginosa culture supernatant compared with the S. Enteritidis culture supernatant. HPLC-MS/MS test demonstrated that AHLs secreted by P. aeruginosa were mainly C14-HSL with a content of 85.71 μg/mL and a small amount of 3-oxo-C12-HSL. In addition, the commercially synthetic C14-HSL had positive effects on the growth of S. Enteritidis, confirming once again that the growth of S. Enteritidis was affected by AHL metabolized by other bacteria and the complexity of bacterial communication.

scholarly journals Proteolytische Aktivität der lysosomalen Enzyme in der Leber wachsender Mäuse

2000 ◽  
Vol 43 (4) ◽  
pp. 363-374
Author(s):  
M. Schmidt ◽  
T. Król ◽  
U. Renne ◽  
L. Panicke
Keyword(s):  
Postnatal Development ◽  
Proteolytic Activity ◽  
Cathepsin D ◽  
Proteolytic Enzymes ◽  
Body Growth ◽  
Male Mice ◽  
High Body

Abstract. Title of the paper: Lysosomal proteolytic activity in the liver of growing mice The behaviour of the activity of some lysosomal proteolytic enzymes in Üie liver of mice, both selected and unselected for high body growth, was followed during the postnatal development. The activity of cathepsin D and L, the alanylaminopeptidase, the arginylaminopeptidase, the α-glucosidase and the N-acetyl-glucosaminidase was estimated in male mice aging 21, 28, 35 and 42 days. In the liver of animals with high body gain statistic significant lower activities (30–50 %) of all estimated enzymes were found, in comparison to the control mice. These results confirm the Statement mat inhibition of proteolysis is an immediate mechanism in the induction of growth.

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