Inhibition, Characterization and Purification of α-AMY from Sera of Iraqi Breast Cancer Patients

Lafta, Namariq A.; Al-Qaisi, Alaa Hussein J.

doi:10.21608/ejhm.2023.286392

Inhibition, Characterization and Purification of α-AMY from Sera of Iraqi Breast Cancer Patients

Document Type : Original Article

Authors

10.21608/ejhm.2023.286392

Abstract

Background: α-Amylase enzyme is digestive enzyme that breaks down complex carbohydrates like starch into simpler sugars; it is being studied as a potential target for anti-cancer therapies.
Objective: The current study aimed to evaluate the general characterization, and inhibition of α-Amylase enzyme, which had been purified from the sera of patients diagnosed with Breast Carcinoma.
Materials and methods: A total of 100 blood serum samples were collected from patients with Breast cancer from Baghdad National Hope Hospital in Baghdad. α-Amylase enzyme was purified from serum clinical isolate in three stages; precipitation with 65% saturated ammonium sulphate, ion exchange chromatography utilizing a DEAE- cellulose column, and gel filtration chromatography with a sephadex G-200 column. Changing conditions in pH, temperature, inhibitor concentration, and kinetics all affect the activity of the α-Amylase enzyme.
Results: α-Amylase inhibition assay showed that Vitis vinifera L. was the most potent inhibitor (71.8%). During the kinetic investigation of the enzyme, it was discovered that the inhibitory mechanism that the extracts use is non-competitive. Vmax was 0.023, and 0.012 mmol min ^-lat 75 μg/ml of Vitis vinifera L. , Verbena Officinalis L .extracts, respectively.
Conclusions: High specific activity for α-Amylase purified from sera of Iraqi breast cancer patients was obtained after three purification steps. Ethanolic plant extracts showed inhibitory effect on α-Amylase while Vitis vinifera L. was the most potent inhibitor.

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