Purification of Single Chain Human Insulin Precursors Using Various Fusion Proteins

  • 박선호 (계명대학교 화학 재료공학부, (주)바이코시스) ;
  • 조정우 (계명대학교 화학 재료공학부) ;
  • 남두현 (영남대학교 약학대학)
  • Published : 2000.11.09

Abstract

For the production of $B^{30}-homoserine$ human insulin precursor, four types of fusion peptides LacZ, MBP, GST, and His-tagged sequence were studied in this work. Recombinant E. coli JM 103 and E. coli JM 109 containing fusion peptides were cultivated at $37^{\circ}C$ for 1hr, and gene expression was occurred when 0.5mM of isopropyl-D-thiogalactoside(IPTG) was added to the culture broth, and followed by longer than 4hr fermentation respectively. DEAE-Sphacel and gel filtration chromatography, amylose and glutathione-Sepharose 4B affinity chromatography, and nickel-affinity chromatography system were employed as purification of $B^{30}-homoserine$ human insulin precursor. Recovery yields of His-tagged, LacZ, GST, and MBP fused $B^{30}-homoserine$ human insulin precursor resulted in 47%, 20%, 20%, and 18%, respectively.

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