Bioprocessing strategies for improving hen egg-white lysozyme (HEWL) production by recombinant Aspergillus niger HEWL WT-13-16

Appl Microbiol Biotechnol. 2002 Dec;60(4):403-7. doi: 10.1007/s00253-002-1122-5. Epub 2002 Nov 5.

Abstract

Hen egg-white lysozyme (HEWL) production by recombinant Aspergillus niger HEWL WT-13-16 from a cDNA under the control of the A. niger glucoamylase promoter was used as a model system. The fungal mycelium was either immobilized on porous Celite 560 micro-carrier or grown in suspension as pelleted and dispersed forms. The objective was to reduce the protease activity that adversely affects the expressed HEWL. Free suspension culture at uncontrolled pH served as the benchmark. The control of pH during growth at pH 4.0 gave rise to a greater than five-fold reduction of protease activity in suspension culture. An additional 38.5% decrease in protease activity was achieved in mycelial-pellet cultures in comparison to a 40.9% decrease in protease activity obtained with Celite 560 beads in an airlift vessel at controlled pH. The specific HEWL yields were 5.8, 5.0 and 4.1 mg/g dry wt. for the free suspension, mycelial-pellet, and Celite-560-immobilized cultures, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aspergillus niger / enzymology
  • Aspergillus niger / genetics*
  • Aspergillus niger / metabolism
  • Bioreactors
  • Biotechnology
  • Cells, Immobilized
  • Chickens
  • Culture Media
  • DNA, Recombinant / genetics
  • DNA, Recombinant / isolation & purification
  • Diatomaceous Earth
  • Hydrogen-Ion Concentration
  • Muramidase / biosynthesis*
  • Muramidase / genetics
  • Ovum / enzymology*
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics

Substances

  • Culture Media
  • DNA, Recombinant
  • Recombinant Proteins
  • Diatomaceous Earth
  • Muramidase