Process for extracellular secretion of Brazzein

A culture medium for manufacturing a Brazzein peptide, the culture medium comprising:

• a nutrient broth comprising dextrose;
• an E. coli cell, the E. coli cell comprising a recombinant nucleic acid, wherein the recombinant nucleic acid comprises:
• (i) a nucleic acid sequence encoding the Brazzein peptide,
• (ii) a nucleic acid sequence encoding a signal leader amino acid sequence, wherein the signal leader amino acid sequence is fused at a 5′ end of the Brazzein peptide, and wherein the signal leader amino acid sequence is operable to direct extracellular secretion of the Brazzein peptide, and
• (iii) an expression control sequence operable to direct expression of the nucleic acid sequence encoding the signal leader amino acid sequence and the nucleic acid sequence encoding the Brazzein peptide;
• and
• an inducing agent.