Question 14.

a. You have seen the structure of sucrose in Lecture 3. The sucrose

disaccharide contains an -1,2-linkage. Shown below are structures of two
isomers of sucrose. Label how the glucosyl and fructosyl units are linked in
each of the isomers.

isomaltulose - -1,6-linkage
trehalulose - -1,1-linkage ( at the fructosyl anomeric carbon)

b. The enzyme sucrose isomerase can catalyze the conversion of sucrose

to isomaltulose and trehalulose. The sucrose isomerase enzyme has
analogies to the glycosidases discussed in Lecture 5. You will need to use
the PyMOL session file Sucrose_isomerase.pse to answer the following
questions. The file shows the Glu254Gln mutant of the enzyme crystallized
with the substrate sucrose.

Is sucrose isomerase more similar to the inverting or retaining

glycosidases? Give one reason based on the structures of the substrate
and products and a second reason based on the enzyme active site.

Retaining glycosidase
The –stereochemistry at C1 of sucrose is retained in the products isomaltulose and trehalulose
The distance between Asp200 and Glu254 is only ~5 Å, a distance typical of a retaining glycosidase.

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c. Draw two mechanisms - One for the conversion from sucrose to
isomaltulose and another for the conversion from sucrose to trehalulose.
Be sure to include any amino acids from the enzyme that are acting as
nucleophiles, general acids or general bases in your mechanisms.

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