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Three extracellular protease-deficient Bacillus subtilisstrains were transformed with the plasmid pCK98 containingthe endo-b-1,4-glucanase (Eng) gene of B. subtilis BSE616.The three transformants, B. subtilis DB104 (pCK98), WB600(pCK98) and WB700 (pCK98), produced the same high levelof enzyme activity and showed similar patterns of cell growthand enzyme production. When B. subtilis DB104 (pCK98), atwo-extracellular protease deficient strain, was cultured for22 h, almost all the secreted enzyme was found to be inthe completely cleaved form by both activity staining andWestern blotting studies. B. subtilis WB600 (pCK98), a sixextracellularprotease-deficient strain, produced a partiallycleaved form in addition to the intact form of the enzyme,although the degree of internal cleavage of the enzyme wasgreatly reduced. With B. subtilis WB700 (pCK98), a sevenextracellularprotease-deficient strain, almost all the enzymewas produced as the intact uncleaved form. This studyillustrates that a role of the Vpr protease is to degrade foreignproteins produced in B. subtilis and WB700 is a suitableexpression system for producing the intact form of the Engand other foreign proteins that may lose at least part of theirefficacy due to internal proteolytic cleavage.

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