- ENZYMATIC ONE-POT REACTION FOR DOUBLE POLYPEPTIDE CONJUGATION IN A SINGLE STEP USING SORTASE

摘要

A method for producing an enzyme conjugation product of three polypeptides comprising the steps of: i) culturing a first polypeptide comprising the amino acid sequence LPXTG (SEQ ID NO: 20, wherein X may be any amino acid residue), the amino acid sequence LPXTA (SEQ ID NO: 31, wherein X may be any amino acid residue), having two N-termini, at its first N-terminus, oligo-glycine _Gm (m = 2 (SEQ ID NO: 22) or 3 (SEQ ID NO: 23), or 4 (SEQ ID NO: 24), or 5 (SEQ ID NO: 25)) and oligo- _{a m (m = 2 (SEQ} ID NO: 26), or 3 (SEQ ID NO: 27) , or 4 (SEQ ID NO: 28) , or 5 (SEQ ID NO: 29) ) of claim 3 having the amino acid sequence Polypeptide, a fourth polypeptide having a sorptase activity derived from Staphylococcus aureus sorghase A, and a fourth polypeptide having a sorbitase activity, and Streptococcus pyogenes sorbetase A Other sorbitan oldest has been reported a method comprising incubating a kinase to a fifth polypeptide having activity at the same time, and recovering the conjugate from the reaction mixture.