Assisted refolding of recombinant prochymosin with the aid of protein disulphide isomerase.
Protein disulphide isomerase (PDI) was shown to be able to accelerate the refolding of unfolded recombinant prochymosin and to enhance the overall yield of active protein. Unlike previous reports in this study PDI was found to be active at pH values as high as 11. The coincidence of the similar apparent optimum pH values of uncatalysed and PDI-catalysed reactions suggests that conditions favourable to spontaneous refolding of proteins may help PDI to catalyse thiol/disulphide interchange. Under the conditions described here no exogenously added dithiothreitol was required for PDI-catalysed renaturation, implying that the disulphide form of PDI was reduced to its active form by the free thiol groups in prochymosin molecules.
Tang
B
B
Institute of Microbiology, Chinese Academy of Sciences, Beijing.
Zhang
S
S
Yang
K
K
D000818:Animals; D002417:Cattle; D012085:Chymosin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004355:Drug Stability; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007535:Isomerases / Q000378:metabolism; D011489:Protein Denaturation / Q000187:drug effects; D019704:Protein Disulfide-Isomerases; D015202:Protein Engineering; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D004792:Enzyme Precursors; D011994:Recombinant Proteins; C032443:prorennin; D012085:Chymosin; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D004229:Dithiothreitol