Expression of human lysozyme in an insoluble form in yeast

Gene. 1987;56(1):53-9. doi: 10.1016/0378-1119(87)90157-0.

Abstract

A high level of expression in yeast of a chemically synthesized human lysozyme (hL) gene was achieved by introducing an A-rich DNA fragment just upstream from the ATG start codon. The synthesized recombinant human lysozyme (r-hL) was insoluble and biologically inactive. It was solubilized with 7 M urea (pH 9) from yeast cells and its lytic activity was efficiently regenerated by oxidative renaturation. This renaturation experiment and Western blotting analysis under reducing and non-reducing conditions indicate that the insoluble form might be caused by the formation of incorrect intra- or intermolecular disulfide bonds. The N-terminal amino acid sequence of the purified r-hL was identical with that of authentic hL.

MeSH terms

  • Amino Acid Sequence
  • Gene Expression Regulation
  • Humans
  • Muramidase / genetics*
  • Muramidase / isolation & purification
  • Plasmids
  • Protein Denaturation
  • Saccharomyces cerevisiae / genetics*
  • Solubility

Substances

  • Muramidase