[Purification of alpha-1,4 leads to 1,4-glucosyltransferase from horse blood serum].

The study is about the extraction and purification process of an enzyme called alpha-1,4-1,4-glucosyltransferase from horse blood serum using various chromatography techniques.

Research Methods

The researchers used different types of ion-exchange chromatography techniques using a series of celluloses (DE-11, DE-32, and CM-32) during the purification process. They carried out the steps in the following order:

• They first applied ion-exchange chromatography on DE-11 cellulose.
• Followed this up with another round of ion-exchange chromatography on DE-32 cellulose.
• Continued the process with ion-exchange chromatography on CM-32 cellulose.
• Finally, the researchers put the substance through gel-filtration on Bio-Gel P-200 for final purification.

Results of the Research

The extracted alpha-1,4-1,4-glucosyltransferase was tested through disc polyacrylamide gel electrophoresis to determine its purity and structure. The results were as follows:

• The protein obtained was determined to be homogeneous, indicating that it is pure.
• The purification degree was approximately 2100 times larger than the initial sample, meaning the concentration of the substance was significantly enhanced during the filtration process.
• Despite the high degree of purification, the yield percentage was only about 40%. This suggests that while the filtration process successfully isolated and purified the enzyme, a significant portion of it was likely lost during the filtration processes.

In conclusion, the research presents an efficient methodology for the purification of alpha-1,4-1,4-glucosyltransferase from horse blood serum. Despite the relative loss in yield percentage, the enrichment degree of the purified enzyme was significant. However, optimization of filtration processes may further increase the yield, enrich the concentration of the enzyme and make the overall process more economical and practical for various biochemical applications.