发明名称 |
Simplified eukaryotic nitrate reductase |
摘要 |
The invention provides modification to the polynucleotide coding sequence for Pichia angusta NAD (P)H: nitrate reductase [YNaR1; GenBank accession number Z49110], which has Enzyme Commission number 1.7.1.2 (formerly EC 1.6.6.2), yielding the polynucleotide coding sequence for simplified eukaryotic nitrate reductase (S-NaR1). The invention also provides a method for recombinant expression of said polynucleotide code in the cells of the methylotrophic yeast Pichia pastoris to produce the polypeptide for S-NaR1, which binds the host-produced molybdenum-molybdopterin cofactor and intracellularly forms catalytically active, nitrate-reducing enzyme as small and stable multimeric proteins. The invention also provides a method for rapid and high-yielding purification of S-NaR1 by utilizing the hexa-histidine sequence at the carboxyl-terminus of said polypeptide for immobilized metal affinity chromatography.
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申请公布号 |
US7262038(B2) |
申请公布日期 |
2007.08.28 |
申请号 |
US20040498697 |
申请日期 |
2004.06.14 |
申请人 |
NITRATE ELIMINATION CO., INC. |
发明人 |
CAMPBELL WILBUR H;BARBIER GUILLAUME G;CAMPBELL ELLEN R |
分类号 |
C12N9/06;C07H21/04;C12N1/19;C12N1/21;C12N15/63;C12N15/81 |
主分类号 |
C12N9/06 |
代理机构 |
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地址 |
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