Sustainable efficient way for opioid peptide LVV-h7 preparation from enzymatic proteolysis in a microfluidic-based reaction-extraction process with solvent recycling

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 May 1:1020:24-8. doi: 10.1016/j.jchromb.2016.03.006. Epub 2016 Mar 15.

Abstract

LVV-h7 (LVVYPWTQFR) is a bioactive peptide that can be obtained from blood as waste of food industry, more precisely from hemoglobin hydrolysis by pepsin. This opioid peptide belongs to the hemorphins family and have strong physiological effects that bring its use in pharmaceutics and various therapeutic treatments attractive, in particular for substituting its costly chemically synthetized analogous. Hemoglobin hydrolysis by pepsin generates a huge variety of peptides among whose LVV-h7 can be purified by liquid-liquid extraction (LLE). Herein, selective preparation of this peptide is proposed by a microfluidic-based continuous reaction-separation process. Hemoglobin hydrolysis in microreactor was firstly coupled to LVV-h7 LLE in octan-1-ol and then coupled to LVV-h7 back LLE in acidic water. This continuous process allowed to prepare pure LVV-h7, as confirmed by liquid chromatography and mass spectrometry. The microfluidic circuit also allowed octan-1-ol recycling in a closed loop, making this method more sustainable than similar biphasic batch process.

Keywords: Bioactive peptide; Enzymatic hydrolysis; Liquid–liquid extraction; Microfluidics; Solvent recycling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bioreactors*
  • Cattle
  • Equipment Design
  • Equipment Reuse
  • Hemoglobins / analysis
  • Hemoglobins / chemistry
  • Hemoglobins / metabolism*
  • Liquid-Liquid Extraction / instrumentation*
  • Liquid-Liquid Extraction / methods
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods
  • Models, Biological
  • Pepsin A / chemistry
  • Pepsin A / metabolism*
  • Peptide Fragments / analysis
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism*
  • Proteolysis

Substances

  • Hemoglobins
  • Peptide Fragments
  • LVV-hemorphin-7
  • Pepsin A