재조합 대장균을 이용한 전달 단백질 rCRM197 생산

초록

Cross reacting materials (CRM197) is a mutant proteinfrom diphtheria toxin (DT) by a single amino acid substitution,glycine to glutamic acid at position 52 (G52E). Inorder to achieve high yields of CRM197, the fermentation processrequires multiple lysogenic mutants of Corynebacteriumdiphtheriae and strictly controlled conditions of growth generally. Wild type CRM197 gene and codon optimized CRM197gene were synthesized to produce recombinant CRM197(rCRM197). Expression vectors were constructed using pQE30,pET-21a, pET-22b and pET-28a for efficient expression by E. coli. Expression conditions such as temperature, induction time,and IPTG concentration were optimized for soluble expressionof rCRM197. In additions, large scale production of rCRM197through the bioreactor, exhibited higher proportion of solubleexpression than flask culture.

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