Patent NZ-601191-A (NZ)
Title: Xylanases, nucleic acids encoding them and methods for making and using them
Inventor: GRAY KEVIN A
IPC: C12N9/42
Filed: 2008-08-01
Abstract: Patent No. 601191 Disclosed is an isolated, synthetic or recombinant nucleic acid comprising a nucleic acid encoding a polypeptide having a xylanase activity, wherein the nucleic acid comprises a sequence having at least 95%, 96%, 97%, 98%, 99%, or more, or complete (100%), sequence identity to SEQ ID NO:1: atggcccagacctgcctcacgtcgccccaaaccggctttcacaatggcttcttctattccttctggaaggacagtccgggcacggtgaatttttgcctgttggagggcggccgttacacatcgaactggagcggcatcaacaactgggtgggcggcaagggatggcagaccggttcacgccggaacatcacgtactcgggcagcttcaatacaccgggcaacggctacctggcgctttacggatggaccaccaatccactcgtcgagtactacgtcgtcgatagctgggggagctggcgtccgccgggttcggacggaacgttcctggggacggtcaacagcgatggcggaacgtatgacatctatcgcgcgcagcgggtcaacgcgccgtccatcatcggcaacgccacgttctatcaatactggagcgttcggcagtcgaagcgggtaggtgggacgatcaccaccggaaaccacttcgacgcgtgggccagcgtgggcctgaacctgggcactcacaactaccagatcatggcgaccgagggctaccaaagcagcggcagctccgacatcacggtgagttga and has at least one mutation selected from (a) to (m), and wherein the said polypeptide shows an improved rate of xylose release when compared to assays with SEQ ID NO: 2: MAQTCLTSPQTGFHNGFFYSFWKDSPGTVNFCLLEGGRYTSNWSGINNWVGGKGWQTGSRRNITYSGSFNTPGNGYLALYGWTTNPLVEYYVVDSWGSWRPPGSDGTFLGTVNSDGGTYDIYRAQRVNAPSIIGNATFYQYWSVRQSKRVGGTITTGNHFDAWASVGLNLGTHNYQIMATEGYQSSGSSDITVS, at the same enzyme loading, and shows an improved rate of xylose release when compared to assays with SEQ ID NO: 2, at a reduced enzyme loading: (a) the nucleic acid codon encoding amino acid residue 38 of SEQ ID NO: 2 is changed from CGT to CAC, and amino acid residue 38 Arginine, Arg (or “R”) is changed to Histidine, His (or “H”); (b) the nucleic acid codon encoding amino acid residue 33 of SEQ ID NO: 2 is changed from CTG to GCG, and amino acid residue 33 Leucine, Leu (or “L”) is changed to Alanine, Ala (or “A”); (c) the nucleic acid codon encoding amino acid residue 73 of SEQ ID NO: 2 is changed from GGC to TAC, and amino acid residue 73 Glycine, Gly (or “G”) is changed to Tyrosine, “Tyr” (or “Y”); (d) the nucleic acid codon encoding amino acid residue 63 of SEQ ID NO: 2 is changed from ATG to GTC, and amino acid residue 63 Isoleucine, Ile (or “I”) is changed to Valine, “Val” (or “V”); (e) the nucleic acid codon encoding amino acid residue 188 of SEQ ID NO: 2 is changed from AGC to GAG, and amino acid residue 188 Serine, Ser (or “S”) is changed to Glutamate, Glu (or “E”); (f) the nucleic acid codon encoding amino acid residue 21 of SEQ ID NO: 2 is changed from TTC to TAC, and the amino acid residue 21 Phenylalanine , Phe (or “F”) is changed to Tyrosine, “Tyr” (or “Y”); (g) the nucleic acid codon encoding amino acid residue 125 of SEQ ID NO: 2 is changed from CAG to TAC, and amino acid residue 125 Glutamine, Gln (or “Q”) is changed to Tyrosine, “Tyr” (or “Y”); (h) the nucleic acid codon encoding amino acid residue 4 of SEQ ID NO: 2 is changed from ACC to CGC, and amino acid residue 4 Threonine, or “Thr” (or “T”) is changed to Arginine, Arg (or “R”); (i) the nucleic acid codon encoding amino acid residue 73 of SEQ ID NO: 2 is changed from GGC to GAG, and amino acid residue 73 Glycine, Gly (or “G”) is changed to Glutamate, Glu (or “E”); (j) the nucleic acid codon encoding amino acid residue 150 of SEQ ID NO: 2 is changed from GTA to GCC, and the amino acid residue 150 Valine, Val (or “V”) is changed to Alanine, Ala (or “A”); (k) the nucleic acid codon encoding amino acid residue 17 of SEQ ID NO: 2 is changed from TTC to GTC, and amino acid residue 17 Phenylalanine , Phe (or “F”) is changed to Valine, “Val” (or “V”); (l) the nucleic acid codon encoding amino acid residue 4 of SEQ ID NO: 2 is changed from ACC to CAC, and amino acid residue 4 Threonine, or “Thr” (or “T”) is changed to Histidine, His (or “H”); and (m) the nucleic acid codon encoding amino acid residue 9 of SEQ ID NO: 2 is changed from CCC to GAC, and the amino acid residue 9 Proline, Pro (or “P”) is changed to Aspartate, Asp (or “D”).