Research article

PARTIAL PURIFICATION OF ALANINE AMINOTRANSAMINASE (ALT) ENZYME IN THE SERA OF GALLSTONE PATIENTS

Manar Mohammed Jassim, Noor Qasim Ahmed, Arwa Saud Alwan

Online First: January 08, 2023


Alanine Amino Transaminase (ALT, GPT) is the plasma enzyme most frequently tested as a sign of damaged liver cells. Even though there is no discernible failure of function, an increase in plasma transaminases activity is a crucial sign of damage to cytoplasmic or mitochondrial membranes. Hepatic ability to synthesize and secrete substances is vast; only severe and typically chronic liver disease can clearly impair prothrombin and albumin synthesis. This study included fifty-five patients with gallstones, their ages ranged between (71-17) years, and forty apparently healthy persons aged between (71-17) years were used as a control group. For the period from the beginning of September to January 2022 in Salah El-Din General Hospital We estimated the activity of the enzyme alanine aminotransferase in the first step, where it was isolated, purified and determined its molecular weight, as well as a kinetic study was conducted for the enzyme, where different methods were used to work and purify this now in the blood by several steps that included precipitation using ammonium sulfate for 50% saturation, which showed a clear increase in purification Molecular enzyme (3.228IU/mg) to (6.664IU/mg) after the first purification step. In the second stage, ion exchange chromatography using DEAE-Cellulose was used in this method to separate the purified enzyme from the sera of people with gallstones, which showed one band, while the molecular weight of the enzyme purified by PAGE, which showed one dense band along the length of the gel was 35 kilo dalton As for the kinetic study of the purified enzyme, it included the optimum conditions for the reaction, which included the concentration of the enzyme 2 mM, and Km= 0.89, pH = 8, and the temperature of 37 °C. incubation time 30 min

Keywords

#