NMR Characterization of the Solution Structure of Acarbose and its Interaction with α-Amylase

Lead Author Major

Biological Sciences

Lead Author Status

Junior

Format

Poster Presentation (Research Day, April 30)

Faculty Mentor Name

Andreas H. Franz

Faculty Mentor Department

Chemistry

Graduate Student Mentor Name

Cate Simmermaker

Graduate Student Mentor Department

Chemistry

Abstract/Artist Statement

Acarbose is a drug used to treat Type II diabetes, a chronic disease indicated by low pancreatic insulin production and poor cellular response to insulin. Acarbose acts as an inhibitor of α-amylase (EC 3.2.1.1), an enzyme involved in the digestion and processing of carbohydrates and starches in the body into the disaccharide maltose, which is further broken down into glucose and used as a source of energy. Treatment with acarbose helps to reduce postprandial hyperglycemia, a symptom of diabetes, by inhibiting the function of α-amylase, effectively decreasing the conversion of starches to sugars and therefore lowering the blood sugar level. In this experiment, the structure of acarbose was assigned by magnetic resonance (NMR) techniques including 1H NMR, 13C NMR, Total Correlation Spectroscopy (TOCSY), Heteronuclear Multiple Bond Correlation (HMBC), and Rotating frame nuclear Overhauser Enhancement (ROESY). The structural topology of binding to α-amylase was studied by Saturation-Transfer Difference (STD) NMR. Inhibition of α-amylase was measured and quantified by conducting an iodine-starch assay, in which iodine indicator is added to solution to visualize intact starch molecules, which form a dark blue complex with iodide. The percent inhibition was measured through the presence/absence of the starch-iodide complex. The results were quantified by UV-VIS absorbance and converted to percent inhibition by comparing the results to standard solutions containing starch and α-amylase.

Location

Information Commons, William Knox Holt Memorial Library and Learning Center

Start Date

30-4-2022 10:00 AM

End Date

30-4-2022 12:00 PM

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Apr 30th, 10:00 AM Apr 30th, 12:00 PM

NMR Characterization of the Solution Structure of Acarbose and its Interaction with α-Amylase

Information Commons, William Knox Holt Memorial Library and Learning Center

Acarbose is a drug used to treat Type II diabetes, a chronic disease indicated by low pancreatic insulin production and poor cellular response to insulin. Acarbose acts as an inhibitor of α-amylase (EC 3.2.1.1), an enzyme involved in the digestion and processing of carbohydrates and starches in the body into the disaccharide maltose, which is further broken down into glucose and used as a source of energy. Treatment with acarbose helps to reduce postprandial hyperglycemia, a symptom of diabetes, by inhibiting the function of α-amylase, effectively decreasing the conversion of starches to sugars and therefore lowering the blood sugar level. In this experiment, the structure of acarbose was assigned by magnetic resonance (NMR) techniques including 1H NMR, 13C NMR, Total Correlation Spectroscopy (TOCSY), Heteronuclear Multiple Bond Correlation (HMBC), and Rotating frame nuclear Overhauser Enhancement (ROESY). The structural topology of binding to α-amylase was studied by Saturation-Transfer Difference (STD) NMR. Inhibition of α-amylase was measured and quantified by conducting an iodine-starch assay, in which iodine indicator is added to solution to visualize intact starch molecules, which form a dark blue complex with iodide. The percent inhibition was measured through the presence/absence of the starch-iodide complex. The results were quantified by UV-VIS absorbance and converted to percent inhibition by comparing the results to standard solutions containing starch and α-amylase.