Renaturation and activation of calf prochymosin produced in an insoluble form in Escherichia coli |
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Authors: | Yoshiyuki Kawaguchi Norio Shimizu Katsuhiko Nishimori Takeshi Uozumi Teruhiko Beppu |
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Abstract: | Calf prochymosin produced in Escherichia coli cells harboring the expression plasmids was insoluble and formed large inclusion bodies, which were solubilized by 8 M urea. The conditions allowing correct refolding of denatured prochymosin were investigated. Dialysis at pH 10 in the presence of 500 mM NaCl was found to give the maximum renaturation, and subsequent acidic treatment for autocatalytic processing of refolded prochymosin allowed almost 100% recovery of chymosin. |
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Keywords: | prochymosin chymosin renaturation |
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