Production of biologically active proteins

A method of preparing a biologically active enzyme composition, said method comprising:

• a) providing a recombinant protein body (PB) assembly formed in an isolated animal cell or multi-cellular fungal cells, wherein the recombinant PB assembly comprises a membrane-enclosed fusion protein, said fusion protein comprising a protein body-inducing sequence (PBIS) and a biologically active enzyme;
• b) contacting the recombinant PB assembly with an aqueous buffer containing a membrane-disassembling amount of a surfactant;
• c) maintaining said contact for a time period sufficient to disassemble the membrane and at a temperature that does not denature the biologically active enzyme, to separate the membrane from the fusion protein to produce a membrane-less recombinant PB assembly that contains the biologically active enzyme;
• d) collecting the membrane-less recombinant PB assembly that contains the biologically active enzyme;
• and
• e) contacting a substrate of the enzyme with the membrane-less recombinant PB assembly, to form the biologically active enzyme composition.