Effect of inhibitor complex formation on the hydration properties of alpha-chymotrypsin. Changes induced in protein hydration by toxylation of the native enzyme

The effect of enzyme-inhibitor complex formation on the hydration properties of the macromolecular moiety was investigated on the model system of α-chymotrypsin and its Ser-195 tosyl derivative. The primary (A-shell) hydration of the native and modified enzyme was compared by sorption measurements. The secondary (B-shell) hydration water was investigated by differential scanning calorimetry. Tosylation is known to induce pronounced conformational changes in the chymotrypsin molecule. These structural modifications have the following effects on the hydration of the native enzyme. The water binding capacity of the protein surface is significantly increased, as shown by both the calorimetric and the sorption results. The amount of unfreezable water of primary hydration is increased by 50 mol H₂O/mol chymotrypsin. The heats (ΔH ) and entropies (ΔS ) of the interaction of water with chymotrypsin are strongly reduced in the modified enzyme. This effect is interpretable by a reduction of the H bonding potential of the protein surface. Parallel to this decrease in δH , the heats of fusion of the secondary hydration water (Q_fus) are significantly increased by tosylation (Q_fus = 256.2 ± 7.8 and 294.2 ± 4.8 J g^?1 H₂O for the native and the tosylated enzyme, respectively). This increase in Q_fus reflects an increase in the extent of H bonding in the B-shell hydration sphere. These changes in the hydration of the native enzyme, associated with the reaction: native chymotrypsin → tosylchymotrypsin, are interpreted by cooperative phase transitions of water molecules in the primary and secondary hydration water. One of these transitions was found to exhibit a significant, linear enthalpy–entropy compensation effect. The compensation temperature \documentclass{article}\pagestyle{empty}\begin{document}$ \hat{\beta} $\end{document} is 290.7 ± 2.8°K. This \documentclass{article}\pagestyle{empty}\begin{document}$ \hat{\beta} $\end{document} value agrees well with compensation temperatures reported in the literature for a series of biochemical reactions in aqueous solution (250–320° K). This agreement in \documentclass{article}\pagestyle{empty}\begin{document}$ \hat{\beta} $\end{document} may point to a common source of both compensation phenomena.     

Polarized fluorescence in an electric field. A model calculation with application to the geometry of the 2-hydroxy-4,4′-diamidinostilbene–DNA complex

Theoretical expressions are derived for the change in the polarized components of the fluorescence, resulting from the orientation of a rigid molecule bearing a chromophore with arbitrary angles for the absorption and transition moments \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _a $\end{document} and \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _e $\end{document} with respect to the molecular axis. The break in the symmetry relation H_V = V_H is related to the tilt angle between \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _a $\end{document} and \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _e $\end{document}. The theory is applied to a sonicated DNA–2-hydroxy-4,4′-diamidinostilbene complex, in the blue and red emission bands of this peculiar dye. Simultaneous measurements of linear dichroism and fluorescence lead to the determination of an angle of 47° between a fluorescent bound dye and the DNA axis, with no difference for the blue- and red-emitting species, but confirm the presence of nonfluorescent bound dye in a more perpendicular arrangement.     

Morphometric diffusing capacity and functional anatomy of the book lungs in the spider Tegenaria spp. (Agelenidae)

The presence of both book lungs and a tracheal system in many spiders raises the question of the functional significance of this double respiratory system. The present physiological and morphometric study of the house spider (Tegenaria spp.) reveals that the diffusing capacity (Dto₂) of the lungs alone suffices during rest and following exercise to meet measured rates of oxygen consumption (\documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm.} $\end{document}o₂) at driving pressures (ΔPto ₂) similar to those calculated for vertebrate lungs. During moulting ΔPto ₂ may rise to more than double the vertebrate values, implying the possible insufficiency of book lungs during this critical life phase. Resting \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ is greatest (92 mm³/h · g) during the early morning and lowest (66 mm³/h · g) near midday: during moulting \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ rises to 278.7 mm³/h · g. In spiders recovering from exercise \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm V}\limits^{\rm .} $\end{document}o₂ is consistently greater than during rest: neither value is significantly reduced by blockage of the tracheal stigmas. Regression calculations of morphometric values for a hypothetical 100-mg Tegenaria yield a total lung volume of 0.578 mm³, a pulmonary surface area of 69.8 mm², and a surface-to-volume ratio of 120.89 mm²/mm³. In spite of the similar thickness of the chitinous and hypodermal components of the air-hemolymph barrier (each ca. 0.2 μm in nonmoulting animals), the low permeability of chitin for oxygen makes this layer the greater barrier to diffusion. For a 100-mg specimen Dto₂ is 3.5 mm³/h · torr: similar to that of a turtle (Pseudemys) on a gram-body weight basis.     

Prognose des Stoffhaushaltes von Staugewässern mit Hilfe kontinuierlicher und semikontinuierlicher biologischer Modelle. II. Prüfung der Prognosegenauigkeit. Prediction of the Balance of Matter in Storage Reservoirs by Means of Continuous or Semicontinuous Biological Models. II. Reliability of the Prediction Method

The phosphate removal in small, completely mixed storage reservoirs (preimpoundment basins) mainly is a function of the production of biomass by the phytoplankton. The knowledge of the critical detention time of the water is the most important premise to the prediction. The critical detention time t̄ is computed from the equation: \documentclass{article}\pagestyle{empty}\begin{document}$ \overline t _c = \frac{1}{{\mu ^* - 0,1}} $\end{document} and the growth rate μ* at a given combination of the light intensity J, temperature T and phosphate concentration P is computed from: \documentclass{article}\pagestyle{empty}\begin{document}$ \mu ^* = \frac{{\mu T \cdot \mu J \cdot \mu P}}{{\mu \max ^2 }}\mu \max \cdot \frac{P}{{K_p + P}}\frac{J}{{K_j + J}}\frac{T}{{T_{opt} }}, $\end{document} (μmax = maximum possible growth rate of the dominant species; K_p, K_j and T_opt are constants computed from batch cultures). The quotient \documentclass{article}\pagestyle{empty}\begin{document}$ \frac{{\bar t_{act.} }}{{\bar t_c }}(\bar t_{act.} = {\rm actual detention time in the water body)} $\end{document} enables prediction of the phosphate removal. A comparison of the predicted results from semicontinuous cultures and from the preimpoundment basin of the Weida reservoir revealed a satisfactory degree of conformity.     

Optical anisotropy of polypeptide chains

Optical anisotropies γ² of N-t-butylacetamide (tBA), N-Methylacetamide (MA), and N, N-dimethylacetamide (DMA) have been determined from the Rayleigh ratios for depolarzed scattering by dilute solutions of the amides in p-dioxane. Traceless optical polarizability tensors \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document} for the amides are derived from these results in conjunction with the Kerr constant for tBA determined by LeGèvre and co-workers. It is shown that the tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document}_i for the glycyle unit in a polypeptide chain may be identified with \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document}MA . Methods for deriving corresponding tensors for other peptide units are indicated and the traceless polarizability tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document} for a polypeptide chain in any specified configuration is formulated.     

Topography of nucleic acid helices in solutions. 3. Interactions of spermine and spermidine derivatives with polyadenylic-polyuridylic and polyinosinic-polycytidylic acid helices

The synthesis of spermine derivatives (II), \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_{\rm 2} {\rm R}_{\rm 3} \mathop {\rm N}\limits^ + \left( {{\rm CH}_2 } \right)_3 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} \left( {{\rm CH}_2 } \right)_2 ]_2 \cdot 4{\rm X}^ - $\end{document}, and spermidine derivatives (III), \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_{\rm 2} {\rm R}_{\rm 3} \mathop {\rm N}\limits^ + \left( {{\rm CH}_2 } \right)_4 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} \left( {{\rm CH}_2 } \right)_3 \mathop {\rm N}\limits^ + {\rm R}_{\rm 1} {\rm R}_{\rm 2} {\rm R}_3 \cdot 3{\rm X}^ - $\end{document}, are reported. The effects of these salts on the helix–coil transition of rA–rU and rI–rC helices were examined. Increasing the size of the hydrophobic substituents, R¹, R², and R³ lowers the degree of stabilization of the helical structure. The disproportionation reaction, 2rA–rU→rA–rU₂ + rA occurs readily with salts II and III, especially when the substituents, R₁, R₂, and R₃ are small, i.e., H or Me. Spermine is found to stabilize the rA–rU² and rI–rC helices to approximately the same extent; however, large differences between the degree of stabilization of rA–rU₂ and rI-rC helices are observed when the substituents R₁, R₂, and R₃ are large hydrophobic groups. Similar results are also obtained for the spermidine series. Finally, differences in the interactions of the salts II and III with rA–rU₂ and rI–rC helices suggest that the latter helix is denser.     

Viscosity of heparin as a function of dielectric constant and of desulfation

The effect of dielectric constant (D) of the solvent on the viscosity of heparin was examined using the relation \documentclass{article}\pagestyle{empty}\begin{document}$ \eta _{{\rm sp}} /c = [\eta ]_\infty (1 + k/\sqrt c) $\end{document}, where [η]_∞ is the shielded intrinsic viscosity obtained by extrapolating \documentclass{article}\pagestyle{empty}\begin{document}$ \eta _{{\rm sp}} /c\,{\rm vs}{\rm . }\,1/\sqrt c ) $\end{document} to infinite concentration, and k is an interaction parameter independent of the dielectric constant of the solvent. This equation was previously reported by the authors⁹ for describing the reduced viscosities of strong polyelectrolytes in salt-free polar solvents. It was found that the [η]_∞ of heparin increases linearly with increasing dielectric constant of the solvent whereas the k values were, within experimental error, independent of D in the range 54.7 < D < 93.2 examined. Graded hydrolysis of heparin from its acid form (heparinic acid) at 57°C resulted in samples of varying degree of desulfation with corresponding decrease in biological activity. It was found that both [η]_∞ and k decrease with increasing desulfation.     

Peptaibol antibiotics: Conformational preferences of synthetic emerimicin fragments

The ir absorption and CD conformational analyses of solutions of the protected 2–9 fragment of the peptaibol antibiotics emerimicins III and IV \documentclass{article}\pagestyle{empty}\begin{document}$ \rlap{--} (Aib_3 \rlap{--} )L - Val - Gly - L - Leu\rlap{--} (Aib_2 \rlap{--} ) $\end{document} and related short sequences are consistent with the presence of a right-handed α-helix for the octapeptide, while the tri-, tetra-, and pentapeptides adopt a 3₁₀-helix, either right- or left-handed, depending on the amino acid sequences. The structural preferences of solid-state \documentclass{article}\pagestyle{empty}\begin{document}$ Z\rlap{--} (Aib_3 \rlap{--} )L - Val - OMe $\end{document} and \documentclass{article}\pagestyle{empty}\begin{document}$ Z\rlap{--} (Aib_3 \rlap{--} )L - Val - Gly - OMe $\end{document} have been determined by x-ray diffraction. In accord with the solution data, incipient 3₁₀-helices, formed by two and three β-turns, have been found for the tetra- and pentapeptides, respectively. The tetrapeptide helix has the left-handed screw sense, while that of the pentapetide is right-handed, thus confirming the conclusions of the CD analysis of the solution.     

Equi-replicated balanced block designs generated by a balanced matrix

In this paper it is shown that if N= \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop \sum \limits_{i = 1}^{S_h} $\end{document} c_ihN_ih, where c_ih are some non-negative integer numbers and N_ih are such incidence matrices that A_h = \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop \sum \limits_{i = 1}^{S_h} $\end{document} i N_ih is a balanced matrix defined by SHAH (1959), for h = 1, 2,…, p, then a block design with an incidence matrix Ñ = [N, N,…,N] is an equi-replicated balanced block design. Here the balance of a block design is defined in terms of the matrix M₀ introduced by CALI?SKI (1971).     

Response to various shock loadings of the constant recycle-sludge-concentration activated-sludge process

The stability of the model of a completely mixed activated-sludge process holding the recycle sludge concentration, X_R, as a system constant subjected to pH, temperature, potassium cyanide, and phenol shock loading was investigated. Soft-drink bottling wastewater was used and maintained at 1000 mg/liter chemical oxygen demand (COD). The hydraulic ratio and recycle sludge concentration were maintained at 0.3 and 7000 mg/liter, respectively. An initial dilution rate of ¼ hr^?1 was maintained for pH and temperature shock loading, with ¼ and ? hr^?1 for KCN shock loading and ¼, ?, and \documentclass{article}\pagestyle{empty}\begin{document}$\frac{1}{16}$\end{document} hr^?1 for phenol shock loading. It was found that the present system could handle pH shock loading as low as 4.0 and as high as 10.4 without any serious disruption of biological solid concentration and filtrate COD. At pH 4.0 shock loading, filamentous organisms were predominant. Temperature shock loading could be handled from 23 to 36°C without any leakage of effluent filtrate COD. At 46°C temperature shock, a 14 hr period was required to recuperate to the new steady state and provided only 85% of COD removal efficiency. For KCN (50 mg/liger) and phenol (85 mg/liter) shock loading, the dilution rates should be lower than \documentclass{article}\pagestyle{empty}\begin{document}$\frac{1}{16}$\end{document} hr^?1 in order to shorten the transient period and improve the effluent quality. Biological kinetic constants included cell yield value, maximum growth rate, and the saturation constant, which was varied with the qualitative shock applied.     

Sequence and age of eruption of deciduous dentition in the baboon (Papio sp)

A detailed eruption sequence and associated age of eruption for deciduous dentition in baboons (Papio sp) are presented in this paper. The sequence was determined by evaluation and comparison of the number and kinds of teeth present in nine age cohorts comprising the study sample of 88 males and 87 females who ranged in age from birth to 763 days. Eruption was assessed visually as present or absent. Several statistical methods used to derive the ages associated with the eruption sequence are described. The basic eruption sequence in the sample population is: i¹ i₁, i₂, i², \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop {\rm c}\limits_{\rm -} {\rm,}\mathop {\rm c}\limits^{\rm -} $\end{document} m¹ (m², m₂), M₁, M¹. Both sexes show the same pattern, with the exception of the second deciduous molar, where males show a sequence of m₂, m₂, while females show the opposite. Posterior dentition shows the greatest gender-specific variation in average age of eruption.     

A Measure of Plankton Community Similarity Utilizing Variable Weighting of Total Density and Taxonomic Proportionality

Four commonly used formulae for measuring percentage similarity (PS) of biological communities were tested for their usefulness in relating to two plankton community properties, species proportional differences and total density differences. The formula best combining species proportionality and total density in the expression of PS is new: where min (xi,yi) is the lesser percentage (doubly standardized) of a species in two samples X and Y and where 2 q, 2xi and 2yi are the total quantities of all species in samples 8,X and Y, where \documentclass{article}\pagestyle{empty}\begin{document}$ \sum\limits_i {z_i } ,\,\sum\limits_i {x_i } \,and\sum\limits_i {y_i } $\end{document} are the total quantities of all species in samples Z, X and Y, respectively. Sample 2 contains the highest density of all species in the set; \documentclass{article}\pagestyle{empty}\begin{document}$ \sum\limits_i {z_i \, > \,(\sum\limits_i {x_i ,\,} \sum\limits_i {y_i } )} $\end{document}. The new expression of PS is simple to use and has the additional advantage of offering the analyst an unlimited choice of weighting factors or importance values for proportionality of species content and total density. The method has been applied to data from Gravenhurst Bay (Ontario) and effectively demonstrates the consequences of phosphorus loading reductions for phytoplankton communities.     

Solution properties of synthetic polypeptides. 18. Helix-coil transition of poly-N5-(2-hydroxyethyl)L-glutamine

The helix–coil transition of poly-N⁵-(2-hydroxyethyl)L -glutamine (PHEG) in aqueous isopropanol was examined by means of optical rotatory dispersion (ORD) and intrinsic viscosity [η] measurements. The Zimm–Bragg parameters σ and s for the transition were determined from the ORD data as a function of molecular weight. It was found that the transition was characterized by a relatively low cooperativity; the values of \documentclass{article}\pagestyle{empty}\begin{document}$ \sqrt \sigma $\end{document} were in the range from 0.039 to 0.066, depending on the solvent composition. These σ values are much larger than those reported for other polypeptide–solvent systems. The transition enthalpy was negative and its magnitude varied with the solvent composition, with a maximum of 620 cal/mol at 40 wt% isopropanol. The curve of [η] versus helical content for a high-molecular-weight sample exhibited a very broad minimum, and this behavior was attributed to the low cooperativity of the transition.     

Depolarized light scattering by amides and peptides

The Rayleigh ratios R_H have been measured for the depolarized scattering dilute solutions of N-ethylacetamide (EA), N-methylpropionamide (MP), and N-acetyl-pyrrolidine (AP) in P-dioxane, and dilute aqueous solutions of N-methylacetamide (MA), N,N-dimethylacetamide (DMA), EA, and MP. Squares of the optical anisot-ropies γ of the amides are obtained through extrapolation of these mesurements to infinite dilution. Values of γ² found for EA and MP in dioxane are in good agreement with calculations based principally on the previously evaluated polarizability tensor of the amide group in conjunction with C? C and C? H bond polarizabilities. The calculations also involve averaging over all conformations, each being weighted according to the estimated conformational energy. The mean-squared optical anisotropies (γ²) of the oligoglycines and oligoalanines are calculated by similarly averaging over all skeletal conformations. The anistropic polarizability tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \hat \alpha $\end{document} for the prolyl structural unit is derived for γ² for AP. The much larger optical anisotropies exhibited by the amides when dissolved in water as compared with those observed in dixane are discussed.     

Thermodynamic functions of biopolymer hydration. I. Their determination by vapor pressure studies,discussed in an analysis of the primary hydration process

The primary hydration process of native biopolymers is analyzed in a brief review of the literature, pertaining to various aspects of biopolymer–water systems. Based on this analysis, a hydration model is proposed that implies that the solution conformation of native biopolymers is stable at and above a critical degree of hydration (h_p′ = 0.06–0.1 g H₂O/g polymer). This water content corresponds to the fraction of strongly bound water, and amounts to ～20% of the primary hydration sphere. In order to test this model, detailed sorption–desorption scanning experiments were performed on a globular protein (α-chymotrypsin). The results obtained are consistent with the proposed hydration model. They show that under certain experimental conditions, sorption isotherms can be obtained that do not exhibit hysteresis. These data represent equilibrium conditions and are thus accessible to thermodynamic treatment. Valid thermodynamic functions, pertinent to the interaction of water with biopolymers in their solution state, can be obtained from these sorption experiments.     

The equivalent conductivity of aqueous salt-free solutions of DNA: Influence of univalent counterions

The equivalent conductivity of salt-free solutions of deoxyribonucleates of alkali metals and ammonium obtained by filtering an isoionic DNA solution through a cation exchanger in the corresponding form has been investigated in the concentrations range of 1 × 10^?4 to 4 × 10^?3M. For all counterions investigated there is a linear dependence of the equivalent conductivity on \documentclass{article}\pagestyle{empty}\begin{document}$ \sqrt {C_p} $\end{document}, where C_p is the nucleic phosphorus concentration. The limiting equivalent conductivity of deoxyribonucleates increases linearly with the limiting mobility of a counterion. By extrapolation to the zero mobility of the counterion, we have obtained the limiting mobility of a macroion, which is equal to 19 × 10^?4 Sm m² equiv.^?1, which is in good agreement with the literature data for denatured DNA obtained by the method of a moving boundary. It is shown that the degree of binding of counterions calculated from the conductometric data in diluted DTA solutions in independent of the nature of the univalent counterion. The degree of dissociation of H⁺-DNA in the isoionic solution calculated with allowance for the fraction of unprotonated bases practically coincides with this value for salts of DNA. The parameter of Manning's theory calculated from the experimental data corresponds to the distance between phosphates along the chain of the macroion, which is equal to 6.7 Å. We attribute the smaller value of this distance as compared with the theoretical one for denatured DNA to the aggregation of macroions.     

Optimization of a stagewise biochemical reactor system with product feedback

A consecutive, first-order, irreversible, biochemical reaction, \documentclass{article}\pagestyle{empty}\begin{document}$ A{\textstyle{{k(\theta)} \over {{\rm Enzyme }1}}} \to B{\textstyle{{k(\theta)} \over {{\rm Enzyme 2}}}} \to C $\end{document}, taking place in a series of N reactors with product recycle is considered. A discrete version of the maximum principle is used to derive general equations necessary for maximizing the production of (1) the final product, C, by choosing the temperature or the pH value in each reactor, and (2) the intermediate product, B, by choosing the reactor volume. A numerical computation for a series of three reactors with recycle is illustrated. The effects of varying the recycle rates on the optimal state and decision variables are also presented.     

Topography of nucleic acid helices in solutions. II. Structure of the double-stranded rA-rU, rI-rC, acid rA, and the triple-stranded rA-rU2 and rA-rI2 helices

Information concerning the structures of rA–rU, rA–rU₂ rI–rC, rA–rI₂, and acid rA helices in solutions is reported. Through the use of diquaternary ammonium salts of the general structure, \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm R}_1 {\rm R}_2 {\rm R}_3 \mathop {\rm N}\limits^ + ({\rm CH}_2 )n\mathop {\rm N}\limits^ + {\rm R}_1 {\rm R}_2 {\rm R}_3 \cdot 2{\rm Br}^ - $\end{document} (I), it is shown that (1) the distances between adjacent negatively charged oxygen atoms on the helix increases in the following order rA–rI₂ < rI–rC < rA–rU ? rA–rU₂; (2) the density of the helices increases in the order. rA–rI₂ < rA–rU < rA–rU₂ < rI–rC; (3) there is a large hydrophobia site in rA–rI₂ and possibly also in rA–rU, rA–rU₂, and rI–rC helices; (4) the results of the interactions between the salts of type I and the helices may be formulated in semi-quantitative terms by the use of two parameters, α, and β which are shown to be related to the charge separation and the density of the helices, respectively; (5) the studies in solutions compare favorably with the x-ray studies on the fibers; and (6) the acid rA helix differs significantly from the other helices by the fact that the electrostatic interstrand interactions between the negatively charged oxygen atom of a phosphate group and the positively charged 10-amino group of adenine contribute significantly to the stabilization of the helix, and thus it is found that the presence of the salts, I, leads to a significant destabilization of the acid rA helix.     

Inhibierungserscheinungen bei der alkoholischen Gärung,I. Einfluß der Ethanolkonzentration auf die spezifische Ethanolbildungsgeschwindigkeit von Saccharomyces cerevisiae

Differential values of the specific ethanol production rate \documentclass{article}\pagestyle{empty}\begin{document}$$ v_{(t)} = \frac{1}{{x_{(t)} }} \cdot \frac{{dP}}{{dt}} $$ \end{document} can be calculated exactly from experimental batch fermentation process data by use of a nonlinear regression programme. The method used is based on the fact, that the function P = f(t) can be approximated by an exponential equation. The specific ethanol production rate is calculated then from the first differential derivation of this equation using the appropriated values of actual biomass concentration. For two strains of Saccharomyces cerevisiae a linear and nonlinear kinetic pattern, respectively, was found for product formation. This result can be explained by a simple mathematical relation according to ν=ν₀ ? a . P^b,in which the exponent becomes 1 in the case of linear kinetic pattern.     

Modellierung der enzymatischen Cellobiosehydrolyse – Vergleich linearer und nichtlinearer Parameterbestimmung

Experimental kinetic data (initial rate and high conversion) on the hydrolysis of cellobiose by 1,4-β-glucosidace (Gliocladium sp.) have been analysed and a competitive inhibition by glucose has been proposed. The determination of kinetic parameters from integral data is based upon algorithms for non-linear optimization and numerical integration. The values of kinetic constants \documentclass{article}\pagestyle{empty}\begin{document}$(v_{\max } = 1.02\frac{{\mu {\rm M}_{{\rm glucose}} }}{{{\rm mg}_{{\rm protein}} \cdot \min }},K_M = 2.6{\rm mM/l, and }K_P = 1.2{\rm mM/l)}$\end{document} agree well with the initialrate results. An important distinction is the confidence limit of parameters. Linear regression analysis shows a virtual accuracy and can lead to wrong conclusions.