scholarly journals H. The Kinetics of Enzyme Cascade Systems General kinetics of enzyme cascades

1969 ◽  
Vol 173 (1032) ◽  
pp. 411-420 ◽  
Keyword(s):  
Product Formation ◽  
Coagulation Cascade ◽  
Lag Phase ◽  
Open Type ◽  
Reaction Velocity ◽  
Cascade Systems ◽  
Enzyme Cascade ◽  
Enzyme Cascades ◽  
Kinetics Of ◽  
Product Relation

The theory of the kinetics of enzyme cascades is developed. Two types of cascades are recognized, one in which the products are stable ( open cascades ) and another in which the products are broken down ( damped cascades ). It is shown that it is a characteristic of a cascade that the final product appears after a certain lag phase. After this lag phase, the velocity of product formation can be very rapid. It is shown that whereas open cascades will always show a complicated time–product relation, damped cascades can under certain circumstances resemble a simple enzymic reaction. Because the relation between the over-all reaction velocity in the extrinsic coagulation cascade and the concentration of any of the proenzymes in this cascade is a hyperbolic one, it is concluded that this cascade is of the damped type rather than the open type.

Presteady-state kinetics of Bacillus 1,3–1,4-β-glucanase: binding and hydrolysis of a 4-methylumbelliferyl trisaccharide substrate

2001 ◽  
Vol 357 (1) ◽  
pp. 195-202
Author(s):  
Mireia ABEL ◽  
Antoni PLANAS ◽  
Ulla CHRISTENSEN
Keyword(s):  
Steady State ◽  
Rate Constant ◽  
Product Formation ◽  
Lag Phase ◽  
Wild Type ◽  
Induced Fit ◽  
Enzyme Substrate ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
Presteady State Kinetics

In the present study the first stopped-flow experiments performed on Bacillus 1,3–1,4-β-glucanases are reported. The presteady-state kinetics of the binding of 4-methylumbelliferyl 3-O-β-cellobiosyl-β-d-glucoside to the inactive mutant E134A, and the wild-type-catalysed hydrolysis of the same substrate, were studied by measuring changes in the fluorescence of bound substrate or 4-methylumbelliferone produced. The presteady-state traces all showed an initial lag phase followed by a fast monoexponential phase leading to equilibration (for binding to E134A) or to steady state product formation (for the wild-type reaction). The lag phase, with a rate constant of the order of 100s−1, was independent of the substrate concentration; apparently an induced-fit mechanism governs the formation of enzyme–substrate complexes. The concentration dependencies of the observed rate constant of the second presteady-state phase were analysed according to a number of reaction models. For the reaction of the wild-type enzyme, it is shown that the fast product formation observed before steady state is not due to a rate-determining deglycosylation step. A model that can explain the observed results involves, in addition to the induced fit, a conformational change of the productive ES complex into a form that binds a second substrate molecule in a non-productive mode.

scholarly journals Kinetics of tau aggregation reveals patient specific tau characteristics among Alzheimer's cases

2021 ◽  
Author(s):  
Tarun V Kamath ◽  
Naomi Klickstein ◽  
Caitlin Commins ◽  
Analiese R Fernandes ◽  
Derek H Oakley ◽  
...  
Keyword(s):  
Growth Phase ◽  
Tau Proteins ◽  
Patient Specific ◽  
Lag Phase ◽  
Plateau Phase ◽  
Kinetic Assay ◽  
Cellular Processes ◽  
Morphological Differences ◽  
Tau Aggregation ◽  
Kinetics Of

Abstract The accumulation of tau aggregates throughout the human brain is the hallmark of a number of neurodegenerative conditions classified as tauopathies. Increasing evidence shows that tau aggregation occurs in a “prion-like” manner, in which a small amount of misfolded tau protein can induce other, naïve tau proteins to aggregate. Tau aggregates have been found to differ structurally among different tauopathies. Recently, however, we have suggested that tau oligomeric species may differ biochemically among individual patients with sporadic Alzheimer disease, and have also showed that the bioactivity of the tau species, measured using a cell-based bioassay, also varied among individuals. Here, we adopted a live-cell imaging approach to the standard cell-based bioassay to explore further whether the kinetics of aggregation also differentiated these patients. We found that aggregation can be observed to follow a consistent pattern in all cases, with a lag phase, a growth phase, and a plateau phase, which each provide quantitative parameters by which we characterize aggregation kinetics. The length of the lag phase and magnitude of the plateau phase are both dependent upon the concentration of seeding-competent tau, the relative enrichment of which differs among patients. The slope of the growth phase correlates with morphological differences in the tau aggregates, which may be reflective of underlying structural differences. This kinetic assay confirms and refines the concept of heterogeneity in the characteristics of tau proteopathic seeds among individuals with Alzheimer’s disease and is a method by which future studies may characterize longitudinal changes in tau aggregation and the cellular processes which may influence these changes.

Effect of Hypertonic Sucrose Upon the Immune Bactericidal Reaction

1970 ◽  
Vol 1 (1) ◽  
pp. 51-55
Author(s):  
Louis H. Muschel ◽  
Linda J. Larsen
Keyword(s):  
Cell Death ◽  
Cell Wall ◽  
Enzymatic Reaction ◽  
Inhibitory Effect ◽  
Lag Phase ◽  
Inner Membrane ◽  
Bactericidal Antibody ◽  
Kinetics Of ◽  
Free Serum ◽  
Supporting Structures

This study was performed to determine the mechanism whereby hypertonic sucrose inhibits the immune bactericidal reaction. Other investigators had postulated that the initial attack of complement (C) on the cell wall was followed with lysozyme-containing whole serum by an enzymatic reaction upon the peptidoglycan substrate resulting in cell death. In the absence of serum lysozyme, secondary lethal changes might occur from damage to the cell's inner membrane as a result of osmotic forces in the presence of a defective cell wall. Hypertonic sucrose giving rise to plasmolysis and protection of the inner membrane was presumed to differentially inhibit the immune response mediated by lysozyme-free serum. The experimental results observed in this investigation have indicated, however, that the inhibitory effect of sucrose upon the bactericidal reaction may be explained simply by its anticomplementary effect and not by any effect on the bacterial cell. This view was supported by the following observations: (i) the comparability of the inhibitory effect of sucrose upon the immune hemolytic and bactericidal reactions, (ii) the comparable percentage loss in bactericidal activity of whole serum and lysozyme-free serum resulting from hypertonic sucrose, (iii) bactericidal antibody titrations were relatively unaffected and C titrations markedly inhibited by sucrose, (iv) the inhibitory effect of sucrose on the bactericidal reaction was unaffected by prior growth of the organism in the presence of sucrose, (v) the kinetics of the bactericidal reactivity of lysozyme-free serum in hypertonic sucrose, compared with whole serum, did not reveal a prolonged lag phase with lysozyme-free serum, but simply diminished reactivity at all times. These observations are compatible with the view that the C attack upon the outer surface of gram-negative bacteria, which plays a part in the cell's permeability control, may account for cell death. In this regard, the immune bactericidal reaction is quite comparable to the lysis of red cells or nucleated cells by C despite the lack of overt lysis in bacteria, probably because of their underlying supporting structures.

scholarly journals Substantiation of the sell-by date of food products

2020 ◽  
Vol 52 (1) ◽  
pp. 59-63
Author(s):  
S.M. Kuzminskiy ◽  
T.V. Adamchuk ◽  
О.М. Holinko ◽  
N.P. Levytska
Keyword(s):  
Food Products ◽  
Storage Conditions ◽  
Lag Phase ◽  
Contamination Level ◽  
Second Phase ◽  
Phase Duration ◽  
Normative Documents ◽  
Culture Development ◽  
Storage Distribution ◽  
Kinetics Of

Objective of the Work. The overview of current methodical approaches for experimental substantiation of the sell-by date of food products. Methods and Materials. Data analysis of scientific literature and normative documents on methods of substantiation of the sell-by date of food products. Results and Discussion. Sell-by date is a period since product’s manufacture, during which it maintains its safety and quality (including nutritional value) within reasonably foreseeable conditions of storage, distribution and consumption. In the case of new products (recipes) introduction it is necessary to review the sell-by date, and its extending as the need arises. The main aspects of microbiological substantiation of the sell-by date of food products are considered. The identification of microbial hazard for particular product is the first phase of the work. The second phase of the work is to determine the kinetic parameters of precise microorganism’s accumulation to maximum permitted level within regulated and aggravated conditions of product’s storage. Conclusions. In the process of microbiological substantiation of the sell-by date of food products it should be taken into consideration the presence of leading pathogen and causative microorganisms of microbial spoilage, the initial contamination level, the lag phase duration of germ culture development, variations between strains, the kinetics of microorganisms’ accumulation within the product in real and aggravated storage conditions, the indetermination connected with biological nature of microorganisms and their inhomogeneous allocation within the product, the limitation for shortcut research methods (if applicable). The decision rule should be based on the consumer’s risk concept. Key Words: food products, sell-by date, substantiation, microbiological indicators.

Experimental and Kinetic Production of Ethanol Using Mucilage Juice Residues from Cocoa Processing

2018 ◽  
Vol 16 (11) ◽  
Author(s):  
Teresa Romero Cortes ◽  
Jaime A. Cuervo-Parra ◽  
Víctor José Robles-Olvera ◽  
Eduardo Rangel Cortes ◽  
Pablo A. López Pérez
Keyword(s):  
Ethanol Yield ◽  
Scale Up ◽  
Pilot Scale ◽  
Product Formation ◽  
Model Parameters ◽  
Pichia Kudriavzevii ◽  
Proposed Model ◽  
Analysis Of Results ◽  
Residual Plots ◽  
Kinetics Of

AbstractEthanol was produced using mucilage juice residues from processed cocoa with Pichia kudriavzevii in batch fermentation. Experimental results showed that maximum ethanol concentration was 13.8 g/L, ethanol yield was 0.50 g-ethanol/g glucose with a productivity of 0.25 g/L h. Likewise, a novel phenomenological model based on the mechanism of multiple parallel coupled reactions was used to describe the kinetics of substrate, enzyme, biomass and product formation. Model parameters were optimized by applying the Levenberg-Marquardt approach. Analysis of results was based on statistical metrics (such as confidence interval), sensitivity and by comparing calculated curves with the experimental data (residual plots). The efficacy of the proposed mathematical model was statistically evaluated using the dimensionless coefficient for efficiency. Results indicated that the proposed model can be applied as a way of augmenting bioethanol production from laboratory scale up to semi-pilot scale.

Kinetics of the IgG—anti-IgG Reaction, as Evaluated by Conventional and Stopped-Flow Nephelometry

1974 ◽  
Vol 20 (8) ◽  
pp. 1071-1075 ◽  
Author(s):  
John Savory ◽  
Gregory Buffone ◽  
Richard Reich
Keyword(s):  
Polyethylene Glycol ◽  
Data Acquisition System ◽  
Lag Phase ◽  
Stopped Flow ◽  
Kinetic Measurement ◽  
Antigen Concentration ◽  
Specific Proteins ◽  
Rapid Reaction ◽  
Antigen Antibody ◽  
Kinetics Of

Abstract A stopped-flow spectrophotofluorometer equipped with a data-acquisition system was used to study the rate of formation of IgG—anti-IgG complexes by nephelometry. Light-scattering aggregates could be detected within 100 ms. Early rates ( 0-3 s) increased with antigen concentration, while rates during later stages (5-100 s) followed the same trend exhibited by the precipitin curve. Polyethylene glycol (known to exert a profound effect on antigen—antibody reactions) at a concentration of 40 g/liter affected the IgG—anti-IgG reaction as follows: there was an initial lag phase of 5-10 s and a subsequent rapid reaction over the next 20-40 s, characterized by a four-to five-fold enhancement in the amount of light scattered. Our observations demonstrate that centrifugal-analyzer techniques can be used for the kinetic measurement of specific proteins in serum.

scholarly journals AKTIVITAS INULINASE OLEH Pichia manshurica DAN FUSAN F4 PADA FERMENTASI BATCH DENGAN UMBI DAHLIA (Dahlia sp) SEBAGAI SUBSTRAT

REAKTOR ◽  
2015 ◽  
Vol 14 (3) ◽  
pp. 187 ◽  
Author(s):  
Wijanarka Wijanarka ◽  
Endang Sutariningsih Soetarto ◽  
Kumala Dewi ◽  
Ari Indrianto
Keyword(s):  
Growth Rate ◽  
Specific Growth Rate ◽  
Specific Growth ◽  
Lag Phase ◽  
Growth Media ◽  
A Value ◽  
Inulinase Activity ◽  
Pichia Manshurica ◽  
Microbial Strains ◽  
Kinetics Of

ACTIVITY OF INULINASE OF Pichia Manshuria AND FUSAN F4 ON BATCH FERMENTATION UDING DAHLIA TUBER (Dahlia sp) AS A SUBSTRATE. A dahlia tuber is one of the common inulin rich crops. Inulin is formed by units of fructans, which are polymers of D-fructose. Inulinases (EC 3.2.1.7) catalyze the hydrolysis of inulin, producing fructooligosaccharides (FOS), inulooligosaccharides (IOS), pulullan, acetone, butanol and sorbitol, therefore dahlia tubers are used as growth media. The inulin hydrolyzing activity has been reported from various microbial strains Pichia manshurica and Fusan F4 which is the result of fusion protoplast. The objective of this study was to determine the activity of inulinase Pichia manshurica and Fusan F4 on the substrate dahlia tubers. Fusan F4 to increase inulinase activity compared with Pichia manshurica and to investigate the kinetics of specific growth rate (μ) and time double (g) from of Pichia manshurica and Fusan F4. The results showed that the exponential phase occurs at 0-12 hour without a lag phase. P. manshurica has a specific growth rate (μ) of 0.18/hour with time double (g) 3.90 hours and the inulinase enzyme activity of 0.56 IU, while for Fusan F4 consecutive has a value μ of 0.20/hour, g of 3.49 hours and the activity of 0.69 IU. The conclusion of this research is to improve Fusan F4 inulinase activity and the ability has to be better than the Pichia manshurica.Umbi dahlia merupakan salah satu umbi yang mengandung inulin. Inulin merupakan polimer fruktan yang dapat dipecah oleh enzim inulinase (E.C. 3.2.1.7) menjadi fruktosa. Fruktosa merupakan bahan baku dasar untuk pembuatan FOS, IOS, pulullan, aseton dan sorbitol, oleh karena itu umbi dahlia digunakan sebagai media pertumbuhan. Enzim inulinase ini secara indigenous dimiliki oleh Pichia manshurica dan Fusan F4 yang merupakan hasil fusi protoplas.Tujuan  penelitian ini adalah  untuk mengetahui aktivitas inulinase Pichia manshurica dan Fusan F4 pada substrat umbi dahlia, Fusan F4 mampu meningkatkan aktivitas inulinase dibandingkan dengan Pichia manshurica serta untuk mengetahui kinetika kecepatan pertumbuhan specifik (µ) dan waktu generasi (g) Pichia manshurica dan Fusan F4. Hasil penelitian menunjukkan bahwa fase  eksponensial terjadi pada jam ke-0 sampai jam ke-12 tanpa diikuti fase lag, Pichia manshurica mempunyai kecepatan pertumbuhan specific (µ)  sebesar 0,18/jam dengan waktu generasi (g) 3,90 jam dan aktivitas enzim inulinase yang dihasilkan sebesar 0,56 IU, sedangkan untuk fusan F4 secara berturut-turut mempunyai nilai µ sebesar 0,20/jam, g sebesar 3,49 jam dan aktivitas sebesar 0,69 IU. Kesimpulan dari penelitian ini adalah Fusan F4 mampu meningkatkan aktivitas inulinase dan mempunyai kemampuan lebih baik dibanding dengan Pichia manshurica.

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