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In vitro DNA binding studies of Aspartame, an artificial sweetener.

Journal of photochemistry and photobiology. B, Biology (2013-02-05)
Soheila Kashanian, Mohammad Mehdi Khodaei, Fahimeh Kheirdoosh
ABSTRACT

A number of small molecules bind directly and selectively to DNA, by inhibiting replication, transcription or topoisomerase activity. In this work the interaction of native calf thymus DNA (CT-DNA) with Aspartame (APM), an artificial sweeteners was studied at physiological pH. DNA binding study of APM is useful to understand APM-DNA interaction mechanism and to provide guidance for the application and design of new and safer artificial sweeteners. The interaction was investigated using spectrophotometric, spectrofluorometric competition experiment and circular dichroism (CD). Hypochromism and red shift are shown in UV absorption band of APM. A strong fluorescence quenching reaction of DNA to APM was observed and the binding constants (Kf) of DNA with APM and corresponding number of binding sites (n) were calculated at different temperatures. Thermodynamic parameters, enthalpy changes (ΔH) and entropy changes (ΔS) were calculated to be +181kJmol(-1) and +681Jmol(-1)K(-1) according to Van't Hoff equation, which indicated that reaction is predominantly entropically driven. Moreover, spectrofluorometric competition experiment and circular dichroism (CD) results are indicative of non-intercalative DNA binding nature of APM. We suggest that APM interacts with calf thymus DNA via groove binding mode with an intrinsic binding constant of 5×10(+4)M(-1).

MATERIALS
Product Number
Brand
Product Description

Supelco
Aspartame, analytical standard
Sigma-Aldrich
bisBenzimide H 33258, for fluorescence, ≥98.0% (HPLC)
Sigma-Aldrich
bisBenzimide H 33258, ≥98% (HPLC and TLC)
Sigma-Aldrich
bisBenzimide H 33258, powder, BioReagent, suitable for cell culture, ≥98% (HPLC and TLC)
Sigma-Aldrich
Asp-Phe methyl ester, ≥98%
Sigma-Aldrich
Hoechst 33258 solution, 1 mg/mL in H2O, ≥98.0% (HPLC)
Aspartame, European Pharmacopoeia (EP) Reference Standard