Methods for Immobilising Enzymes

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  • Enzyme molecules mixed with immobilising support.
  • They bind to it due to a combination of hydrophobic interactions and ionic links.
  • Adsorbing agents include: porous carbon, glass beads, clays and resins.
  • Weak bonding forces can lead to leakage (detachment of enzyme molecules.)
  • Attaching enzymes without denaturing and displaying the active site leads to high reaction rates.
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Covalent Bonding:

  • Enzyme molecules covalently bonded to support.
  • Covalently linked enzymes are linked to an insoluble material using a crosslinking agent (gluteraldehyde or sepharose.)
  • Does not immobilise large quantities of enzyme.
  • Strong binding so leakage is small.
  • Covalent bonding can denature actibe site.
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  • Enzymes may be trapped in gel bead or cellulose fibre network.
  • Trapped in natural state so active site will not be affected.
  • Reduced reaction rates as substrate must pass through barrier to reach active site and form enzyme-substrate complex.
  • Active site less easily available than adsorption or covalent bonding.
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Membrane Separation:

  • Enzymes and substrates may be separated by a partially permeable membrane. 
  • Enzymes are held on one side of membrane, substrate passed along opposite side.
  • Enzyme molecules are too large to pass through membrane.
  • Substrate molecules are small enough to pass through membrane.
  • Product molecules are small enough to pass back through the membrane.
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