In industrial biotechnology, enzymes can be immobilised using different methods. One common technique is entrapment within a polymer gel matrix, such as calcium alginate.
Which of the following describes a major limitation specifically associated with the entrapment method of enzyme immobilisation?
The active sites of the enzymes are commonly deactivated or blocked because covalent bonds form between the amino acid R-groups and the support matrix.
Large substrate molecules face significant diffusion limitations through the matrix, making this method unsuitable for reactions involving macromolecular substrates.
The enzymes are highly prone to leaching into the product stream because they are only held by weak hydrophobic and electrostatic interactions.
The entrapped enzyme molecules become more flexible, resulting in a significantly lower thermal stability compared to free enzymes in solution.
