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Figure 5 | Microbial Cell Factories

Figure 5

From: Pre-expression of a sulfhydryl oxidase significantly increases the yields of eukaryotic disulfide bond containing proteins expressed in the cytoplasm of E.coli

Figure 5

Production of BPTI in the cytoplasm of E. coli. Analysis of mature BPTI wild-type and mutants which mimic the two disulfide folding intermediates produced in the cytoplasm of origami with pre-expression of Erv1p and PDI in LB media at 30°C. Panel A) Coomassie stained SDS-PAGE. T = total E. coli lysate; S = soluble fraction. The position of BPTI is marked with an arrow. Panel B) rpHPLC analysis of purified BPTI mutants under conditions which allow identification of the different folding intermediates. Upper trace (displaced 50 mAU) C30A/C51A mutant, lower trace C14A/C38A mutant. A single peak with the elution point being equivalent to the relevant two disulfide containing wild-type folding intermediate was obtained for both proteins.

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Microbial Cell Factories

ISSN: 1475-2859

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