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Model of the regulation of activity of immobilized enzymes (amylases) in soil

Content Provider	Springer Nature Link
Author	Demkina, E. V. Shanenko, E. F. Nikolaev, Yu. A. El’ Registan, G. I.
Copyright Year	2017
Abstract	The preservation of activity of extracellular enzymes in soil is presently associated with their immobilization on organic or inorganic carriers. Enzyme immobilization results, however, in a significant decrease in enzymatic activity. In the present work, the mechanism responsible for promotion of the catalytic activity was revealed, as well as the favorable effect of low-molecular alkylhydrozybenzenes of the class of alkylresorcinols, which are common in soil organic matter, on stability of immobilized enzymes (exemplified by amylases) by their post-translational modification. Optimal conditions (enzyme to sorbent ratio, pH optimum, CaCl$_{2}$ concentration, and sorption time) for amylase sorption on a biological sorbent (yeast cell walls) were determined and decreased activity of the immobilized enzyme compared to its dissolved state was confirmed. Alkylresorcinols (C$_{7}$AHB) at concentrations of 1.6 to 80 mM were found to cause an increase of amylase activity both in the case of already sorbed enzymes (by 30%) and in the case of a free dissolved enzyme with its subsequent immobilization (by 50–60%). In both cases, the optimal C$_{7}$AHB concentration was 16 mM. Amylase stability was determined for C7AHB-modified and unmodified enzymes immobilized on the biological sorbent after two cycles of freezing (–20°C) and thawing (4°C). Inverse dependence was revealed between increasing stability of C$_{7}$AHB-modified enzymes and an increase in their activity, as well as higher stability of immobilized modified amylases than of the dissolved modified enzyme. Investigation of the effect of C$_{7}$HOB-modification in the preservation of activity in immobilized amylases after four freeze–thaw cycles revealed: (1) better preservation of activity by the modified immobilized enzymes compared to immobilized ones; (2) differences in the dynamics of activity loss within compared pairs, with activity of immobilized amylases decreasing after the second cycle to a lower level (42%) than activity of the modified immobilized enzymes after the fourth cycle (48%). These results demonstrate that in the preservation of activity of extracellular enzymes in soil both stabilization mechanisms are of importance: immobilization on organic carriers and modification of the enzyme conformation by low-molecular compounds with the functions of chemical chaperones.
Starting Page	231
Ending Page	240
Page Count	10
File Format	PDF
ISSN	00262617
Journal	Microbiology
Volume Number	86
Issue Number	2
e-ISSN	16083237
Language	English
Publisher	Pleiades Publishing
Publisher Date	2017-04-04
Publisher Place	Moscow
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	extracellular soil enzymes stability freeze–thawing activity immobilization conformational modifications alkylresorcinols Microbiology Medical Microbiology
Content Type	Text
Resource Type	Article
Subject	Microbiology Applied Microbiology and Biotechnology

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