Secretion of human interleukin-2 in biologically active form by Bacillus brevis directly into culture medium.
We constructed an efficient system for synthesis and secretion of human interleukin-2 (IL-2) by Bacillus brevis. The secretion vector we constructed had strong promoters and contained the region coding for the signal peptide of the gene for B. brevis 47 cell-wall protein, followed directly by the gene encoding mature IL-2. Modification of the signal peptide and use of a protease-deficient mutant of B. brevis HPD31 increased productivity. When the signal peptide was more basic near its amino terminal and more hydrophobic in the middle region, IL-2 production increased 20 fold. Production by the mutant harboring the secretion vector was four fold that of the parent harboring the same plasmid. The yield of IL-2 increased further to 0.12 g/liter, when cultural conditions were made optimal, such by the addition of Tween 40 to the medium. The IL-2 produced by B. brevis had the same biological activity as authentic IL-2. Biologically active human IL-2 was produced efficiently and secreted directly into the medium by B. brevis.
Takimura
Y
Y
Department of Applied Biological Sciences, Faculty of Agriculture, Nagoya University, Japan.
Kato
M
M
Ohta
T
T
Yamagata
H
H
Udaka
S
S
D000595:Amino Acid Sequence; D001407:Bacillus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D003470:Culture Media; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000172:deficiency; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D021382:Protein Sorting Signals / Q000096:biosynthesis / Q000235:genetics
D003470:Culture Media; D007376:Interleukin-2; D021382:Protein Sorting Signals; D004247:DNA; D010450:Endopeptidases