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    Simplified eukaryotic nitrate reductase
    3.
    发明申请
    Simplified eukaryotic nitrate reductase 失效
    简化的真核硝酸还原酶

    公开(公告)号：US20060211084A1

    公开(公告)日：2006-09-21

    申请号：US10498697

    申请日：2002-12-10

    申请人： Wilbur Campbell ,  Guillaume Barbier ,  Ellen Campbell

    发明人： Wilbur Campbell ,  Guillaume Barbier ,  Ellen Campbell

    IPC分类号： C12N15/74 ,  C07H21/04 ,  C12P21/06 ,  C12N9/06 ,  C12N1/18

    CPC分类号： C12Q1/26 ,  C12N9/0044 ,  C12N15/81 ,  G01N33/84 ,  G01N2333/906

    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.

    摘要翻译： 本发明提供了对毕赤酵母NAD（P）H：硝酸还原酶[YNaR1; GenBank登录号Z49110]，其具有1.7.1.2的酶委员会（以前的EC 1.6.6.2），产生简化的真核硝酸还原酶（S-NaR1）的多核苷酸编码序列。 本发明还提供了一种用于在甲基营养酵母巴斯德毕赤酵母细胞中重组表达所述多核苷酸编码的方法，以产生用于S-NaR1的多肽，其结合宿主产生的钼钼卟吩辅因子并在细胞内形成催化活性的硝酸盐还原 酶作为小而稳定的多聚体蛋白。 本发明还提供了通过利用所述多肽的羧基末端的六组氨酸序列来固定金属亲和色谱来快速和高产纯化S-NaR1的方法。

    Production of N-acylated sulphur-containing amino acids with microorganisms having enhanced N-acyltransferase enzymatic activity
    5.
    发明授权
    Production of N-acylated sulphur-containing amino acids with microorganisms having enhanced N-acyltransferase enzymatic activity 失效
    用具有增强的N-酰基转移酶酶活性的微生物生产N-酰化的含硫氨基酸

    公开(公告)号：US08143031B2

    公开(公告)日：2012-03-27

    申请号：US12370434

    申请日：2009-02-12

    申请人： Rainer Figge ,  Guillaume Barbier ,  Gwénaëlle Bestel-Corre

    发明人： Rainer Figge ,  Guillaume Barbier ,  Gwénaëlle Bestel-Corre

    IPC分类号： C12N1/15 ,  C12N1/19

    CPC分类号： C12P13/12 ,  C12N9/1029

    摘要： The present invention claims an isolated polypeptide having L-amino-acid-N-acyl transferase enzymatic activity and a modified microorganism in which this enzyme is overexpressed. Substrates of said enzyme include mainly methionine and their derivatives or analogs. Overexpression in sulphur-containing amino acid producing microorganisms permits the production of large amounts of N-acylated sulphur-containing amino acids. The isolation of the N-acylated sulphur-containing amino acids from the fermentation medium is also claimed.

    摘要翻译： 本发明要求具有L-氨基酸-N-酰基转移酶酶活性的分离多肽和其中该酶过表达的经修饰的微生物。 所述酶的底物主要包括甲硫氨酸及其衍生物或类似物。 含硫氨基酸生成微生物中的过表达允许产生大量的N-酰化含硫氨基酸。 还要求从发酵培养基中分离N-酰化的含硫氨基酸。