Novel a-galactosidases

A method of modifying a red blood cell, comprising:

• obtaining a group B or group AB erythrocyte, suspending it in a buffer solution having a neutral pH, and contacting it a polypeptide having the sequence specified as:
• SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:
• 5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8 or SEQ ID NO:
• 9, thereby cleaving the branched immunodominant B epitopes on the group B or group AB erythrocyte as determined by serological typing, or hemagglutination assays.

A purified enzyme, comprising:

• a polypeptide including the sequence specified by SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:
• 5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8 or SEQ ID NO:
• 9, wherein the enzyme displays alpha galactosidase branched substrate specificity and a neutral pH optima.

A purified enzyme, comprising:

• (a) a polypeptide having at least 10 amino acids in the following sequence numbered accordingly when aligned with SEQ ID. NO:2:
• M at residue 10;
• G at residue 47;
• G at residue 84;
• Y at residue 86;
• Y at residue 99;
• N at residue 102;
• K at residue 114;
• T at residue 127;
• G at residue 130;
• G at residue 132;
• G at residue 139;
• N at residue 156;
• D at residue 160;
• P at residue 164;
• G at residue 205;
• R at residue 277;
• R at residue 281;
• F at residue 287;
• G at residue 308;
• Q at residue 312;
• I at residue 317;
• R at residue 333;
• D at residue 340;
• G at residue 346;
• G at residue 349;
• G at residue 360;
• D at residue 363;
• D at residue 364;
• N at residue 367;
• H at residue 369;
• G at residue 370;
• T at residue 371;
• G at residue 396;
• E at residue 462;
• N at residue 463;
• T at residue 465;
• T at residue 467;
• P at residue 468;
• R at residue 483;
• G at residue 484;
• L at residue 486;
• T at residue 489;
• N at residue 498;
• 1 at residue 508;
• D at residue 513;
• W at residue 517;
• E at residue 519;
• G at residue 521;
• D at residue 525;
• 1 at residue 528;
• N at residue 531;
• F at residue 533;
• I at residue 549;
• P at residue 553;
• I at residue 573;
• A at residue 590;
• G at residue 595;
• N at residue 601;
• and, I at residue 629;
• and
• (b) wherein the polypeptide has at least 20% identity with SEQ ID NO:
• 2, and wherein the polypeptide has α3 galactosidase activity.

A method of producing a recombinant enzyme, comprising:

• obtaining a nucleic acid encoding SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:
• 5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, or SEQ ID NO:
• 8;
• expressing the nucleic acid in a cell transfected therewith;
• inducing expression of the nucleic acid encoding the enzyme;
• and purifying the expressed enzyme from the cell, wherein the enzyme has α3 galactosidase activity.

A non-naturally occurring prokaryotic cell, comprising:

• an expression vector not found in a wild-type prokaryotic cell, the expression vector having a nucleic acid sequence encoding a polypeptide including the sequence specified by SEQ ID NO:
• 2, SEQ ID NO:
• 3, SEQ ID NO:
• 4, SEQ ID NO:
• 5, SEQ ID NO:
• 6, SEQ ID NO:
• 7, SEQ ID NO:
• 8 or SEQ ID NO:
• 9, and wherein the polypeptide has α3 galactosidase activity.