Nitrilases, nucleic acids encoding them and methods for making and using them

1. An isolated, synthetic or recombinant nucleic acid, encoding a polypeptide having nitrilase activity, comprising

(a) a nucleic acid having a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or complete identity (100% identical) to SEQ ID NO:209; or

(b) a nucleic acid encoding a polypeptide comprising amino acids having a sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or complete identity (100% identical) to SEQ ID NO:210; or

(c) a nucleic acid completely complementary to (a) or (b).

2. A nucleic acid vector capable of replication in a host cell, wherein the vector comprises the nucleic acid of claim 1 .

3. A host cell comprising the nucleic acid of claim 1 .

4. A host organism comprising the host cell of claim 3 .

5. The isolated, synthetic or recombinant nucleic acid of claim 1 , wherein the nucleic acid is affixed to a solid support, wherein optionally the solid support is selected from the group of a gel, a resin, a polymer, a ceramic, a glass, a microelectrode and any combination thereof.

6. A method for making a polypeptide having nitrilase activity, the method comprising

(a) introducing the nucleic acid of claim 1 encoding the polypeptide into a host cell under conditions that permit production of the polypeptide by the host cell, and

(b) recovering the recombinant polypeptide so produced.

7. A kit comprising the nucleic acid of claim 1 , or a fragment thereof encoding a polypeptide having nitrilase activity, and a buffer.

8. The isolated, synthetic or recombinant nucleic acid of claim 1 , having at least 95% sequence identity to SEQ ID NO:209.

9. The isolated, synthetic or recombinant nucleic acid of claim 1 , having at least 98% sequence identity to SEQ ID NO:209.

10. The isolated, synthetic or recombinant nucleic acid of claim 1 , having at least 99% sequence identity to SEQ ID NO:209.

11. The isolated, synthetic or recombinant nucleic acid of claim 1 , further consisting of one or more mutations, wherein:

nucleotides 163-165 of SEQ ID NO:209 are substituted with AAA, AAG, GGT, GGC, GGA, GGG, CAA, or CAG;

nucleotides 178-180 of SEQ ID NO:209 are substituted with GAA or GAG;

nucleotides 331-333 of SEQ ID NO:209 are substituted with TCT, TCC, TCA, TCG, AGT, or AGC;

nucleotides 568-570 of SEQ ID NO:209 are substituted with CAT, CAC, TCT, TCC, TCA, TCG, AGT, AGC, ACT, ACC, ACA, TCA, TAT, TAC, ATG or ACG;

nucleotides 571-573 of SEQ ID NO:209 are substituted with TTA, TTG, CTT, CTC, CTA, CTG, GTT, GTC, GTA, GTG, ATG, ACT, ACC, ACA, GAT, GAC, GGT, GGC, GGA, GGG, GAA, GAG, TAT, TAC, or ACG;

nucleotides 595-597 of SEQ ID NO:209 are substituted with GAA, GAG, TTA, TTG, CTT, CTC, CTA, or CTG;

nucleotides 664-666 of SEQ ID NO: 209 are substituted with TTA, TTG, CTT, CTC, CTA, or CTG; or any combination thereof.