11. As part of a research project in biochemistry you are charged with purifying
ID: 149890 • Letter: 1
Question
11. As part of a research project in biochemistry you are charged with purifying a dehydrogenase enzyme from a species of bacteria. Your advisor informs you that four isoforms of enzyme exist in the cell: three soluble isoforms and a membrane-associated isoform. Two of the soluble isoforms occur naturally as monomers while the other functions as a homo-tetramer. Analysis of the genes for the monomeric forms indicates significant differences in their respective amino acid compositions, most notably in the percentages of the following amino acids (Glu, Asp, Thr). Generate an outline of the purification scheme you would utilize to isolate the more soluble of the monomeric enzyme isoforms. For each methodology employed provide a chemically based rationale for its inclusion, i.e., briefly explain how each step gets you closer to a pure solution of the desired enzyme. (10) a.Explanation / Answer
Please find the answers below:
Answer a: In order to purify the monomeric and heteromeric soluble and non-soluble membrane-bound isoforms, follwing steps can be performed:
Answer b: In order to understand the biochemical nature of the proteins, bioinformatics approach is required to be followed. A top-down technique must be performed to know the genetic sequece pertaining to this protein. Further, this sequence can be provided in online available tools and searched for analogy and homology run. This will give an estimation of similarities between the sequences among different species. The particular presence and sequence of amino acids in the proteins will be highly useful in this process.
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