Recombinant flounder growth hormone from Escherichia coli: overexpression, efficient recovery, and growth-promoting effect on juvenile flounder by oral administration.
An efficient production method for recombinant flounder growth hormone (r-fGH) from Escherichia coli was developed and the biological activity of purified r-fGH was examined using juvenile flounder. The use of bicistronic construction in the expression plasmid resulted in the production of over 40% of the E. coli cellular protein as r-fGH. The r-fGH was recovered from cell lysates following inclusion body washing, solubilization and refolding in sodium dodecylsulfate (SDS) solution, and removal of contaminated proteins with secondary butanol treatment. The SDS content in purified r-fGH solution was adjusted to appropriate levels by diafiltration. More than 47% of the r-fGH was recovered from the E. coli cell lysates and the purity of recovered r-fGH was 98%. The oral administration of purified r-fGH to juvenile flounder, once a week for 4 weeks at a dosage of 40 micrograms r-fGH g-1 fish body weight, resulted in significant increases both in weight and length. These results of overexpression, simple purification with high recovery yield and purity, and good growth-promoting activity of the r-fGH suggest that the production scheme described in this study is useful for the potential application of r-fGH in fish farming.
Jeh
H S
HS
Biotech Research Institute, LG Chem Research Park, Yu Song, Science Town, Taejon, South Korea.
Kim
C H
CH
Lee
H K
HK
Han
K
K
D000284:Administration, Oral; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003545:Cysteine / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics*; D005432:Flounder / Q000235:genetics / Q000254:growth & development*; D015870:Gene Expression; D013006:Growth Hormone / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification
D011994:Recombinant Proteins; D013006:Growth Hormone; D003545:Cysteine