THE PROTEOLYTIC ENZYMES OF MICROORGANISMS: VII. A STUDY OF SOME OF THE PROPERTIES OF TWO PROTEASES ISOLATED FROM MORTIERELLA RENISPORA DIXON-STEWART

1954 ◽  
Vol 32 (1) ◽  
pp. 60-67 ◽  
Author(s):  
L. R. Wetter
Keyword(s):  
Enzymatic Activity ◽  
Glutamic Acid ◽  
Culture Medium ◽  
Proteolytic Enzymes ◽  
Isoelectric Points

Two proteases isolated from the culture medium of Mortierella renispora Dixon-Stewart (PRL 26) had different isoelectric points but similar pH optimums. The components were inhibited by equal amounts of ovomucoid. Heat studies showed that the constituent moving towards the cathode (cathodic component) was more stable at 58 °C. than the anodic component. The enzyme located in the cathodic component hydrolzyed chloroacetyl-L-tyrosine. The same fraction also hydrolyzed N-carbobenzoxy-α-L-glutamyl-DL-alanine and to a much lesser extent N-carbobenzoxy-α-L-glutamyl-L-glutamic acid. No enzymatic activity could be detected against glycylglycine, diglycylglycine, glycyl-L-proline, seryl-serine, or α-benzoyl-L-argininamide.

THE PROTEOLYTIC ENZYMES OF MICROORGANISMS: VII. A STUDY OF SOME OF THE PROPERTIES OF TWO PROTEASES ISOLATED FROM MORTIERELLA RENISPORA DIXON-STEWART

1954 ◽  
Vol 32 (1) ◽  
pp. 60-67 ◽  
Author(s):  
L. R. Wetter
Keyword(s):  
Enzymatic Activity ◽  
Glutamic Acid ◽  
Culture Medium ◽  
Proteolytic Enzymes ◽  
Isoelectric Points

Two proteases isolated from the culture medium of Mortierella renispora Dixon-Stewart (PRL 26) had different isoelectric points but similar pH optimums. The components were inhibited by equal amounts of ovomucoid. Heat studies showed that the constituent moving towards the cathode (cathodic component) was more stable at 58 °C. than the anodic component. The enzyme located in the cathodic component hydrolzyed chloroacetyl-L-tyrosine. The same fraction also hydrolyzed N-carbobenzoxy-α-L-glutamyl-DL-alanine and to a much lesser extent N-carbobenzoxy-α-L-glutamyl-L-glutamic acid. No enzymatic activity could be detected against glycylglycine, diglycylglycine, glycyl-L-proline, seryl-serine, or α-benzoyl-L-argininamide.

Zur Genese Der Amidisch An Den Chromophor Von Pyoverdinen Gebundenen Dicarbonsäuren [1]

1991 ◽  
Vol 46 (5-6) ◽  
pp. 398-406 ◽  
Author(s):  
H. Schäfer ◽  
K. Taraz ◽  
H. Budzikiewicz
Keyword(s):  
Glutamic Acid ◽  
Dicarboxylic Acid ◽  
Succinic Acid ◽  
Pseudomonas Fluorescens ◽  
Exponential Growth ◽  
Culture Medium ◽  
Culture Time ◽  
Hydrolysis Of

Pseudomonas strains of the so-called fluorescent group usually produce several pyoverdins which differ only in the nature of a dicarboxylic acid bound amidically to the chromophor. For the pyoverdins isolated from the culture medium of Pseudomonas fluorescens 12 it is shown that succinic acid is an artefact formed by hydrolysis of succinic amide, and that a-ketoglutaric acid is transformed enzymatically to glutamic acid. This process is reversed after the phase of exponential growth of the bacteria. The ratio C4- vs. C5 -acids changes with the culture time and with increasing Fe3+ content of the medium in favor of the latter

THE PROTEOLYTIC ENZYMES OF MICROORGANISMS: III. SOME CHARACTERISTICS OF EXTRACELLULAR PROTEASES PRODUCED IN SUBMERGED CULTURE

1950 ◽  
Vol 28c (6) ◽  
pp. 600-612 ◽  
Author(s):  
W. B. McConnell
Keyword(s):  
Low Temperatures ◽  
Free Amino ◽  
Submerged Culture ◽  
Culture Medium ◽  
Proteolytic Enzymes ◽  
Amino Groups ◽  
Extracellular Proteases ◽  
Egg Albumin ◽  
Rapid Hydrolysis ◽  
Hydrolysis Of

Some of the general characteristics of the proteases liberated into the culture medium by molds and actinomycetes grown in submerged culture have been studied. Species of Alternaria, Streptomyces, Mortierella, and Gliocladium were used. The enzymes resemble trypsin in that they are most active at a pH slightly above 7 and are inhibited by a preparation of egg albumin. They are stable at low temperatures but suffer marked losses in activity when stored for 16 hr. above 40 °C. The most rapid hydrolysis of gelatin occurs at temperatures between 40 °C. and 50 °C. The enzymes from different organisms show definite differences with respect to their ability to attack different proteins, gelatin and casein being in general the most readily digested. The protease systems from different organisms also vary with respect to the extent to which they can digest gelatin; some enzymes are able to release about three times as many amino groups from gelatin as others. The limit of the hydrolysis is not dependent upon substrate concentration but is slightly affected by the concentration of enzyme. The enzymes were effective in liberating free amino acids from gelatin.

scholarly journals Disease-related Metabolites in Culture Medium of Fibroblasts from Patients with d-2-Hydroxyglutaric Aciduria, l-2-Hydroxyglutaric Aciduria, and Combined d/l-2-Hydroxyglutaric Aciduria

2003 ◽  
Vol 49 (7) ◽  
pp. 1133-1138 ◽  
Author(s):  
Eduard A Struys ◽  
Nanda M Verhoeven ◽  
Birthe Roos ◽  
Cornelis Jakobs
Keyword(s):  
Cell Culture ◽  
Glutamic Acid ◽  
Cell Lines ◽  
Culture Medium ◽  
Culture Media ◽  
Control Cell ◽  
Fibroblast Cell ◽  
Gas Chromatography Mass Spectrometry ◽  
Organic Acidurias ◽  
Stable Isotope Dilution

Abstract Background: d-2-Hydroxyglutaric aciduria (D-2-HGA), l-2-hydroxyglutaric aciduria (L-2-HGA), and the combined d/l-2-hydroxyglutaric aciduria (D/L-2-HGA) are poorly understood organic acidurias. To investigate the usefulness of cultured human skin fibroblasts for both diagnostic and research purposes, we measured disease-related metabolites in the cell culture medium. Methods: We measured d-2-hydroxyglutarate (D-2-HG), l-2-hydroxyglutarate (L-2-HG), succinate, 2-ketoglutarate, and citrate in fibroblast cell medium by stable-isotope-dilution gas chromatography–mass spectrometry and glutamine, glutamic acid, and lysine with an amino acid analyzer. We used six cell lines from patients with D-2-HGA, two from patients with L-2-HGA, three from patients with D/L-2-HGA, and seven control cell lines. Culture medium was analyzed after a 96-h incubation period. Results: Culture media from cell lines from D-2-HGA patients contained D-2-HG at concentrations 5- to 30-fold higher than media from controls, whereas the concentration of L-2-HG in media was not increased. Media from L-2-HGA cell lines showed a fivefold increase in L-2-HG compared with controls. Media containing fibroblasts from D/L-2-HGA patients contained moderately increased amounts of both D-2-HG and L-2-HG. For all cell lines, succinate concentrations in the blank medium were higher than after 96 h of incubation with the exception of two of three D/L-2-HGA cell lines. Media of D-2-HGA cell lines had 2-ketoglutarate concentrations that were 40% of that for controls. Glutamic acid concentrations in media of these cell lines were 60% lower than in controls. Conclusions: Cell culture media from fibroblasts from patients with D-2-HGA, L-2-HGA, or D/L-2-HGA contain increased amounts the corresponding 2-HGs, demonstrating the suitability of fibroblasts for both diagnosis of and research concerning 2-HGAs.

Endo-1, 4-β-glucanase and β-glucosidase of the ciliate Polyplastron multivesiculatum free of cellulolytic bacteria

1986 ◽  
Vol 32 (3) ◽  
pp. 219-225 ◽  
Author(s):  
A. Bonhomme ◽  
G. Fonty ◽  
M. J. Foglietti ◽  
D. Robic ◽  
M. Weber
Keyword(s):  
Enzymatic Activity ◽  
Isoelectric Focusing ◽  
Reducing Sugars ◽  
Proteolytic Enzymes ◽  
Cellulose Degradation ◽  
Protein Fractions ◽  
Intracellular Bacteria ◽  
Cellulolytic Bacteria ◽  
Derivatives Of ◽  
Bacterial Cellulase

To determine the contribution of protozoal activity to cellulose degradation, we maintained the ciliate Polyplastron multivesiculatum free of extracellular and intracellular cellulolytic bacteria. Control experiments to verify the absence of such bacteria were performed on cellular extracts, on filtrates, and on ciliates before lyophilization. The enzymatic activity was determined by viscometry and by determining the amount of reducing sugars produced. The enzyme was found to be an endo-1, 4- β-glucanase. Polyplastron multivesiculatum which was maintained free of extracellular and intracellular bacteria degraded soluble derivatives of cellulose and slightly degraded native cellulose. The activity was not due to intracellular bacterial cellulase, as ingested bacteria were lysed and digested by proteolytic enzymes of the protozoan. In addition, when preparing the filtrate solutions, P. multivesiculatum was maintained in culture for 5 days with Streptococcus faecalis (a facultatively anaerobic, noncellulolytic bacterium). Isoelectric focusing and chromatofocusing of lyophilized P. multivesiculatum that was free of cellulolytic bacteria yielded three protein fractions that degrade carboxymethylcellulose and hydroxyethylcellulose. Chromatofocusing also revealed the presence of two protein fractions with β-glucosidase activity.

scholarly journals Critical role of glutamic acid 202 in the enzymatic activity of stromelysin-1 (MMP-3)

2001 ◽  
Vol 268 (3) ◽  
pp. 826-831 ◽  
Author(s):  
Begoña Arza ◽  
Marc De Maeyer ◽  
Jordi Félez ◽  
Désiré Collen ◽  
H. Roger Lijnen
Keyword(s):  
Enzymatic Activity ◽  
Glutamic Acid ◽  
Critical Role

Synthesis and Relaxometric Characterization of a MRI Gd-Based Probe Responsive to Glutamic Acid Decarboxylase Enzymatic Activity

2013 ◽  
Vol 56 (6) ◽  
pp. 2466-2477 ◽  
Author(s):  
Roberta Napolitano ◽  
Giorgio Pariani ◽  
Franco Fedeli ◽  
Zsolt Baranyai ◽  
Markus Aswendt ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Glutamic Acid ◽  
Glutamic Acid Decarboxylase ◽  
Acid Decarboxylase

scholarly journals BACTERIAL FORMATION OF L-GLUTAMIC ACID FROM ACETIC ACID IN THE GROWING CULTURE MEDIUM

1961 ◽  
Vol 7 (1) ◽  
pp. 30-40 ◽  
Author(s):  
TOSHINAO TSUNODA ◽  
ISAMU SHIIO ◽  
KOJI MITSUGI
Keyword(s):  
Acetic Acid ◽  
Glutamic Acid ◽  
Culture Medium

scholarly journals Isolation and some properties of extracellular amylases from Fusarium martii var. minus f.2

2014 ◽  
Vol 16 (2) ◽  
pp. 237-245 ◽  
Author(s):  
Joanna Boguszewska
Keyword(s):  
Enzymatic Activity ◽  
Active Center ◽  
Liquid Culture ◽  
Culture Medium ◽  
Sh Groups ◽  
Liquid Culture Medium

α-amylase and glucoamylase were isolated from the liquid culture medium of <i>Fusarium martii</i>. At 10<sup>-3</sup> M concentrations PCMB inhibited the activity of both enzymes, which indicated that they require SH groups for enzymatic activity, α-amylase was also inhibited by DFP at 10<sup>-3</sup> M. One may thus assume that serine is present in the active center of the examined amylase.

Proteolytic activity of Oidiodendron kalrai

1975 ◽  
Vol 21 (9) ◽  
pp. 1362-1368 ◽  
Author(s):  
P. M. Cino ◽  
R. P. Tewari
Keyword(s):  
Basement Membrane ◽  
Enzymatic Activity ◽  
Human Serum ◽  
Proteolytic Activity ◽  
Crude Extract ◽  
Protease Activity ◽  
Proteolytic Enzymes ◽  
Cell Free Extract ◽  
Yeast Phase ◽  
Biological Substrates

The physiochemical characteristics of the intracellular proteolytic enzymes of Oidiodendron kalrai, a neuropathogenic fungus, were studied. The organism in the yeast phase was grown in a semisynthetic medium containing 1% tryptone, at 37 °C for 48 h, on a gyrotory shaker. The crude extract was prepared by breaking the cells in a French pressure cell and the proteolytic activity was tested against biological substrates. The cell-free extract hydrolyzed casein, hemoglobin, lactalbumin, gelatin, elastin, collagen, and purified rabbit renal basement membrane to various degrees. Optimal proteolytic activity was observed at pH 6 and at 32 °C. Calcium and EDTA did not affect the enzymatic activity; however, activity was partially inhibited by sulfhydryl-blocking agents and by heat-inactivated horse, calf, and human serum. The extract was totally inactivated by exposure to a temperature of 70 °C for 60 min. Storage at −76 °C or −15 °C for 6 months or at 4 °C for 4 weeks did not affect protease activity.

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