Direct high-level secretion into the culture medium of tuna growth hormone in biologically active form by Bacillus brevis.
The characteristic features of the Bacillus brevis system developed by us are very high productivity of heterologous proteins and very low extracellular proteinase activity. However, the production level of eucaryotic proteins with this system was generally one or two orders of magnitude lower than that of bacterial proteins. Therefore, we have explored methods for increasing the production efficiency as to animal proteins. Signal peptide modification was found to be very effective for high-level secretion of tuna growth hormone (tGH). Modification of the signal peptide with higher basicity in the amino terminal region and higher hydrophobicity in the middle region brought about a ten-fold increase in tGH production. Further elevation of the tGH yield to 240 mg/l was achieved by using a low proteinase mutant and a stable plasmid, and by culturing B. brevis under optimal conditions with the addition of some chemicals. Thus, biologically active tGH can be efficiently produced directly in the medium with this B. brevis system.
Sagiya
Y
Y
Department of Applied Biological Sciences, Faculty of Agriculture, Nagoya University, Japan.
Yamagata
H
H
Udaka
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000235:genetics*; D001483:Base Sequence; D001709:Biotechnology; D003001:Cloning, Molecular; D003470:Culture Media; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014413:Tuna / Q000235:genetics*
D003470:Culture Media; D018076:DNA, Complementary; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D013006:Growth Hormone