Introduction the Principle of Saturation Additionality for Enzymatic Process into the Methodology of Plants Raw Materials Complete Processing

Background: Biotechnological approach to complete processing of plant raw materials using enzyme preparations allows the efficient use of its native biological and/or technological potential. The point components are fragments of molecular components of the cell wall matrix with a target glycoside bonds' hard-to-find concentration which is necessary to determine the enzyme kinetic characteristics. 
Materials and methods: Non-granulated dry non-molassed sugar beet pulp as well as lyase and hydrolase enzymes were used as research objects Purpose: to study an approach based on approximation of experimental data followed by determination of horizontal asymptotes. 
Results: The adequacy of using an indirect indicators, such as specific electrical conductivity, for assessing the kinetic parameters of lyase and hydrolase enzymes  is substantiated. An array of experimental data was obtained for dynamics of specific electrical conductivity during processing beet pulp with enzyme preparations in the concentration range from 0 to 0.8%. As a result of the approximation, the local limits of the substrate concentration were calculated in indirect units. For dynamic of the "substrate - enzyme" system, a set of postulates was proposed. On the basis of this set, the principle of saturating complementarity for enzymatic process is formed. In according with it the local substrate concentration limit achieved at a given enzyme concentration is a fractional part of some global one, which can be completely converted into a product through several stages where the local limit of the duration of each stage tends to infinity. It has been experimentally established that, in the case of lyase and hydrolase enzyme, the calculated local substrate concentration limits increase monotonously with an increase of enzyme concentration, degenerating into a horizontal asymptote corresponding to the global substrate concentration limit, which confirms both the principle of saturating complementarity and its consequences. The applicability of low enzyme concentrations (within 0.1-0.2%) and several step-by-step stages for the enzymatic transformation of a substrate into a product has been experimentally established. 
Conclusion: As a result of research, the principle of saturating complementarity for enzymatic process was developed. Based on experimental data on the enzymatic transformation dynamics for processed object, it allows an unambiguous determination of the effective substrate concentration in the absence of certainty about its quantity value, or the impossibility of direct determination. The resulting value can be used to establish the kinetic characteristics of enzymatic process, such as Vmax and Km. The principle of saturating complementarity is applicable for lyase and hydrolase homoenzymes. It is a one of the necessary components within the decision tree for development the technologies for industrial production of plant polyglycans.

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