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KMID : 0380019940090020165
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Korean Journal of Biotechnology and Bioengineering
1994 Volume.9 No. 2 p.165 ~ p.172
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The Structural Characterizaion of Recombinant Bovine Somatotropin Expressed in Escheirchia coli
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Kim, Jungho/김정호
Kim, Hoonjoo/Park, Eun Suk/Kim, Joon/김훈주/박은숙/김준
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Abstract
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In this paper we have described the structural characterization of recombinant bovine somatotropin produced in Escherichia colt. Recombinant bovine somatotropin consists of 191 amino acid residues with a calculated molecular weight of 21,802 Da. For fragmentation of recombinant bovine somatotropin, we have used trypsin, Staphylococcus aureus V8 protease, CNB;mild acid hydrolysis method. Digestion and cleavage with these proteases and chemicals yielded peptides of various size for amino acid sequence determination. The N-terminal sequence analysis was carried out up to thirty residues. Because the design of the recombinant bovine somatotropin gene for expression was such that the coding sequence begins with an initiation codon, AUG, before Ala, the first amino acid of bovine somatotropin, we could expect the initial amino acid as N-formyl Met. But the first amino acid of this protein, expressed in E. colt cells as inclusion bodies, was Ala. And the amino acid composition of RP-HPLC purified recombinant bovine somatotropin was determined and no essencial difference was observed. The amino acid sequence of the recombinant bovine somatotropin was identical to that predicted from its recombinant gene. There was no processing or replacement of amino acid residues in recombinant bovine somatotropin expressed in E. coli. The hydropathy plot of recombinant bovine somatotropin revealed a hydrophobic region at the NH:,-terminus and hydrophilic region at the COOH-terminus. The E. colt expression system is thought to be valuable for the expression of recombinant bovine somatotropin because protein was processed to remove the N-terminal Met residue by methionyl-aminopeptidase autonomously.
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