Controlling the Kinetics of an Enzymatic Reaction through Enzyme or Substrate Confinement into Lipid Mesophases with Tunable Structural Parameters

Int J Mol Sci. 2020 Jul 20;21(14):5116. doi: 10.3390/ijms21145116.

Abstract

Lipid liquid crystalline mesophases, resulting from the self-assembly of polymorphic lipids in water, have been widely explored as biocompatible drug delivery systems. In this respect, non-lamellar structures are particularly attractive: they are characterized by complex 3D architectures, with the coexistence of hydrophobic and hydrophilic regions that can conveniently host drugs of different polarities. The fine tunability of the structural parameters is nontrivial, but of paramount relevance, in order to control the diffusive properties of encapsulated active principles and, ultimately, their pharmacokinetics and release. In this work, we investigate the reaction kinetics of p-nitrophenyl phosphate conversion into p-nitrophenol, catalysed by the enzyme Alkaline Phosphatase, upon alternative confinement of the substrate and of the enzyme into liquid crystalline mesophases of phytantriol/H2O containing variable amounts of an additive, sucrose stearate, able to swell the mesophase. A structural investigation through Small-Angle X-ray Scattering, revealed the possibility to finely control the structure/size of the mesophases with the amount of the included additive. A UV-vis spectroscopy study highlighted that the enzymatic reaction kinetics could be controlled by tuning the structural parameters of the mesophase, opening new perspectives for the exploitation of non-lamellar mesophases for confinement and controlled release of therapeutics.

Keywords: alkaline phosphatase; confined diffusion; controlled release; drug delivery; enzymatic kinetics; liquid crystalline mesophases; phytantriol.

MeSH terms

  • Alkaline Phosphatase / chemistry*
  • Biocatalysis
  • Enzymes, Immobilized / chemistry*
  • Fatty Alcohols / chemistry
  • Hydrophobic and Hydrophilic Interactions
  • Kinetics
  • Lipids / chemistry*
  • Liquid Crystals / chemistry*
  • Models, Molecular
  • Nitrophenols / chemistry
  • Organophosphorus Compounds / chemistry
  • Substrate Specificity
  • Sucrose / analogs & derivatives
  • Sucrose / chemistry

Substances

  • Enzymes, Immobilized
  • Fatty Alcohols
  • Lipids
  • Nitrophenols
  • Organophosphorus Compounds
  • sucrose monostearate
  • nitrophenylphosphate
  • Sucrose
  • 3,7,11,15-tetramethyl-1,2,3-hexadecanetriol
  • Alkaline Phosphatase