Production of the human-beta-defensin using Saccharomyces cerevisiae as a host.
Studies of the human defensins have been hampered by the lack of a simple expression system allowing for rapid production of functional peptide forms. Here, we describe a Saccharomyces cerevisiae AH22 expression system that meets that condition. The 42 amino acid form of human beta-defensin-1 was expressed under the control of the ADH1 promoter. The optimum conditions for expression were determined and the stable maintenance of the pVT103L-hBD-1 chimeric vector in the yeast population was confirmed. Expressed hBD-1 was secreted into the medium (approximately 55 microg l(-1)) and purified using cation-exchange chromatography. Isolated defensin exhibited strong bactericidal effect on Escherichia coli ML-35p. We conclude that the expression system described here will be a useful tool where readily prepared and active forms of the human defensins are needed.
Cipáková
Ingrid
I
CPN spol. s r.o., 561 02 Dolní Dobrouc 401, Czech Republic. icipakova@yahoo.com
Hostinová
Eva
E
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D023083:beta-Defensins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D023083:beta-Defensins