Nitrile hydratase variant

An isolated nitrile hydratase comprising an α-subunit and a β-subunit, wherein:

• (a)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 36, 148 and 204 of SEQ ID NO:
• 1 to Met, Asp and Arg respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 41, 51 and 108 of SEQ ID NO:
• 2 to Ile, Val, and Asp respectively;
• (b)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 6, 36 and 126 of SEQ ID NO:
• 1 to Thr, Met and Tyr respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 10, 118, and 200 of SEQ ID NO:
• 2 to Asp, Val and Glu respectively;
• (c)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 19, 71 and 126 of SEQ ID NO:
• 1 to Val, His and Tyr respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 37, 108, and 200 of SEQ ID NO:
• 2 to Leu, Asp, and Glu respectively;
• (d)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 19, 71 and 126 of SEQ ID NO:
• 1 to Val, His and Tyr respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 37, 108, and 200 of SEQ ID NO:
• 2 to Val, Asp, and Glu respectively;
• or
• (e)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 148 and 204 of SEQ ID NO:
• 1 to Asp and Arg respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 108, and 200 of SEQ ID NO:
• 2 to Asp, and Glu respectively.

An isolated nitrile hydratase comprising an α-subunit and a β-subunit, wherein:

• (a) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1;
• and
• (b) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions;
• 10 to Asp, Glu, Trp, Gly, Tyr or Cys,
• 32 to Gly,
• 37 to Thr, Ala, Leu, Ile or Val,
• 41 to Glu, Thr, Ala, Leu, Ile, or Val,
• 46 to Gly, Tyr, Leu ,Lys or Asp,
• 48 to Gly, Ala, Val, Ser, Thr or Arg,
• 51 to Ala or Val,
• 72 to Phe,
• 118 to Ala, Leu, Ile or Val,
• 127 to Ala, Val or Ser,
• 146 to Gly,
• 160 to Leu or Trp,
• 186 to Glu, Asp, Lys, Arg, Asn, Ser or Gly
• 217 to Gly,
• 160 and 186 to Trp and Arg respectively,
• 127, 160 and 186 to Ser, Trp and Arg respectively,
• 51 and 108 to Val and Asp respectively,
• 118 and 200 to Val and Glu respectively,
• 10, 118 and 200 to Asp, Val and Glu respectively,
• 37, 108 and 200 to Leu, Asp and Glu respectively,
• 37, 108 and 200 to Val, Asp and Glu respectively,
• 41, 51 and 109 to Ile, Val and Asp respectively,
• 46, 108 and 212 to Lys, Arg and Tyr respectively, or
• 48, 108 and 212 to Val, Arg and Tyr respectively.

An isolated nitrile hydratase comprising an α-subunit and a β-subunit, wherein:

• (a)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 6, 19 and 126 of SEQ ID NO:
• 1 to Thr, Val and Tyr respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 46, 108 and 212 of SEQ ID NO:
• 2 to Lys, Arg and Tyr respectively;
• (b)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 6, 19 and 126 of SEQ ID NO:
• 1 to Thr, Val and Tyr respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 48, 108 and 212 of SEQ ID NO:
• 2 to Val, Arg and Tyr respectively, or
• (c)(i) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at positions 6, 19 and 126 of SEQ ID NO:
• 1 to Ala, Val and Tyr respectively, and (ii) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at positions 127, 160 and 186 of SEQ ID NO:
• 2 to Ser, Trp and Arg respectively.

An isolated nitrile hydratase comprising an α-subunit and a β-subunit, wherein:

• (a) the α-subunit comprises the polypeptide of SEQ ID NO:
• 1, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 1 is substituted at one or more positions selected from the group consisting of positions 36, 71, 148, and 204 by another amino acid;
• and
• (b) the β-subunit comprises the polypeptide of SEQ ID NO:
• 2, except that the amino acid sequence of the polypeptide of SEQ ID NO:
• 2 is substituted at one or more positions selected from the group consisting of positions 10, 32, 37, 41, 46, 48, 51, 72, 118, 127, 146, 160, 186, and 217 by another amino acid.