Efficient secretion of human lysozyme fused to the Sh ble phleomycin resistance protein by the fungus Tolypocladium geodes.
Tolypocladium geodes strain NC50 was transformed by different integrating vectors bearing both a synthetic gene encoding human lysozyme (HLz) and the Sh ble phleomycin resistance marker, either in separate expression cassettes or in transcriptional or translational fusion configurations. Clones derived from all vectors were able to secrete HLz. The highest productivities in shake flasks (up to 150 mg l-1 in 5 days) were obtained when HLz was fused at the C-terminal end of the Sh ble protein. The fusion protein is efficiently secreted and release of active lysozyme occurs by extracellular proteolytic cleavage in the junction peptide.
Laboratoire de Microbiologie et Génétique appliquées du CNRS, CRBGC, Université Paul Sabatier, Toulouse, France.
M
M
Baron
G
G
Tiraby
T
T
Calmels
M
M
Parriche
H
H
Durand
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000138:chemical synthesis; D004352:Drug Resistance, Microbial / Q000235:genetics; D005069:Evaluation Studies as Topic; D005822:Genetic Vectors; D006801:Humans; D003904:Mitosporic Fungi / Q000235:genetics; D008969:Molecular Sequence Data; D009113:Muramidase / Q000235:genetics / Q000378:metabolism*; D010692:Phleomycins / Q000494:pharmacology*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping; D014170:Transformation, Genetic
D010692:Phleomycins; D011993:Recombinant Fusion Proteins; D004247:DNA; D009113:Muramidase