Enzyme Extraction and Purification from Chicken Liver - Biochemistry Lab Report

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ENZYME EXTRACTION AND PURIFICATION FROM CHICKEN LIVER
Results
Malate dehydrogenase is one of the key components in a aspartate aminotransferase, AST, kit for
diagnosis, a kit that is often quite sensitive when it comes to taking measurements of the enzyme
concentration of AST in blood, plasma as well as serum that is normally set as a marker test for
the functioning of the lover in the checking, diagnosis and treatment of diseases related with the
liver. The procedure for purification of MDH is quite convenient as it encompassed ammonium
sulfate fractionation, exclusion of size chromatography on Sephacryl S-300 resin s well as anion-
exchanger DEAE-cellulose chromatography of resin1.
Various methods for MDH purification were outlined from the chicken liver using ion-exchange
chromatography with affinity elution using NADH. The pattern of the chromatograph of the
chicken liver MDH on the DEAE-cellulose resign illustrated the existence e of a key MDH
enzyme activity peak which labelled chicken liver MDH eluted using 0.01 M NaCl and single
minor peak illustrated the activity of MDH to have been resolved and eluted using 0.05 M NaCl.
Collection, concentration and placement onto the Sephacryl S-300 column of the major peak
fractions that illustrated the activity of MDH was done and was as illustrated in the figure which
demonstrated the appearance of the peak of a single MDH enzyme activity2.
1 Couée, Ivan, and Keith F. Tipton. "Glutamate dehydrogenase." In Glutamine and Glutamate Mammals, pp. 81-
100. CRC Press, (2018)
2 Darwish, Doaa A., Hassan MM Masoud, Mohamed M. Abdel-Monsef, Mohamed S. Helmy, and Mahmoud A.
Ibrahim. "Purification and characterization of malate dehydrogenase from sheep liver (Ovis aries): Application in
AST assay diagnostic kit." Journal of Applied Pharmaceutical Science Vol8, no. 02 (2018)

The specific activity of the MDH was raised to about 6.7 units/gram protein in the subsequent
size-exclusion chromatography which stood for 11.6 fold purification as well as about 28%
output. An avalanche of purification fold as well as per cent for recovery for MDH was reported.
The MDH from the chicken liver was found to be having 225-fold with 47% production.
The graph of the corrected absorbance versus the time could be plotted as illustrated below