ISSN:
1040-0397
Keywords:
Ferrocene-amylose
;
Depolymerization
;
α-Amylase
;
Human saliva
;
Activity assay
;
Cyclic voltammetry
;
Rotating disk electrode
;
Chemistry
;
Polymer and Materials Science
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Chemistry and Pharmacology
Notes:
The behavior of (ferrocene)amylose (FA), in the presence of amylolytic depolymerases (α-amylase from Aspergillus oryzae and human saliva), has been investigated by cyclic voltammetry at a rotating disk electrode (CVA/RDE). Growth of the limiting current with time in the presence of the enzymes is proportional to the amount of enzyme introduced. The quantitative data treatment to assay the endoamylolytic activity of enzymes at CVA/RDE involves plotting (idt/id0)4.5 against time; the slope of the linear plot being equal to (rate) Mn0C-1, where idt and id0 are the limiting currents at time t and 0, respectively, (rate) is the enzymatic activity, Mn0 is the number averaged molecular weight of FA at t = 0, and c is its concentration. The comparison of CVA/RDE with the 3,5-dinitrosalicylic acid and the Somogyi-Nelson reducing saccharides procedures shows advantages of the former, especially in assaying small quantities of enzymes. Also the CVA/RDE approach is simpler and takes place under much milder conditions. The main disadvantage of CVA/RDE is the inhibiting effect of Triton X-100 in the reaction between FA and the amylases which is not observed in the case of native, ferrocene-free amylose. In general, CVA/RDE appears to be an attractive analytical method for monitoring diverse enzymatic depolymerization reactions.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elan.1140090803
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