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The holoenzyme is denatured by heat at very high temperature. It is found that two polypeptides, although denatured, contains chains that remain together as a unit, while two other denatured polypeptides are floating free. You interpret this to mean that:
I. The primary structure of the subunits changed during the high temperature heat treatment.
II. A disulfide bridge was present linking only two of the subunits in holoenzyme.
III. Ionic bonds, hydrogen bonds, and hydrophobic interactions were disrupted during the heat treatment.
All the efforts that go into the preparation of instruments are futile if the sterilization process itself is not successful. There is no way of seeing that instruments are sterile
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