公开(公告)号：US07115403B1

公开(公告)日：2006-10-03

申请号：US09571553

申请日：2000-05-16

申请人： Frances H. Arnold ,  Ioanna P. Petrounia ,  Lianhong Sun

发明人： Frances H. Arnold ,  Ioanna P. Petrounia ,  Lianhong Sun

IPC分类号： C12N9/02 ,  C12N1/20 ,  C12N15/00 ,  C12Q1/68 ,  C12Q1/00 ,  C07H21/04 ,  C12P21/04

CPC分类号： C12N9/0006 ,  C12N15/102

摘要： This invention relates to the expression of improved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly oxidase enzymes. The enzymes are advantageously produced in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this manner include improved expression, enhanced activity toward one or more substrates, and increased thermal stability. In a particular embodiment, the invention relates to improved expression of the galactose oxidase gene and galactose oxidase enzymes. GAO mutants that are highly active and/or thermostable are disclosed.

摘要翻译： 本发明涉及编码真核生物酶，特别是氧化酶的改良的多核苷酸和多肽序列的表达。 酶有利地在常规或简单的表达系统中产生。 可以使用用于多核苷酸序列的定向进化的各种方法来获得改进的序列。 以这种方式产生的多肽或蛋白质的改进特征包括改进的表达，增强对一种或多种底物的活性和增加的热稳定性。 在一个具体实施方案中，本发明涉及半乳糖氧化酶基因和半乳糖氧化酶的改进表达。 公开了高活性和/或热稳定的GAO突变体。

公开(公告)号：US07098010B1

公开(公告)日：2006-08-29

申请号：US09722602

申请日：2000-11-27

申请人： Frances H. Arnold ,  Ioanna P. Petrounia ,  Lianhong Sun

发明人： Frances H. Arnold ,  Ioanna P. Petrounia ,  Lianhong Sun

IPC分类号： C12N9/02 ,  C12N1/20 ,  C12N15/00 ,  C12Q1/68 ,  C12Q1/00 ,  C12P21/04 ,  C07H21/04

CPC分类号： C12N9/0006 ,  C12N15/102

摘要： This invention relates to the expression of improved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly oxidase enzymes. The enzymes are advantagoeusly produced in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this manner include improved expression, enhanced activity toward one or more substrates, and increased thermal stability. In a particular embodiment, the invention relates to improved expression of the galactose oxidase gene and galactose oxidase enzymes. GAO mutants that are highly active and/or thermostable are disclosed.

摘要翻译： 本发明涉及编码真核生物酶，特别是氧化酶的改良的多核苷酸和多肽序列的表达。 这些酶在常规或简单的表达系统中有利地产生。 可以使用用于多核苷酸序列的定向进化的各种方法来获得改进的序列。 以这种方式产生的多肽或蛋白质的改进特征包括改进的表达，增强对一种或多种底物的活性和增加的热稳定性。 在一个具体实施方案中，本发明涉及半乳糖氧化酶基因和半乳糖氧化酶的改进表达。 公开了高活性和/或热稳定的GAO突变体。

公开(公告)号：US07220563B2

公开(公告)日：2007-05-22

申请号：US10375909

申请日：2003-02-27

申请人： Frances H. Arnold ,  Lian-Hong Sun ,  Ioanna P. Petrounia

发明人： Frances H. Arnold ,  Lian-Hong Sun ,  Ioanna P. Petrounia

IPC分类号： C12N9/02 ,  C12N1/20 ,  C12N15/00 ,  C12Q1/00 ,  C12Q1/68 ,  C12P21/04 ,  C07H21/04

CPC分类号： C12N9/0006 ,  C12Y101/03004 ,  C12Y101/03009

摘要： Glucose oxidase enzymes are provided, including novel variants of galactose oxidase enzymes. The polynucleotides that encode these novel variants can be expressed in recombinant host cell expression systems. The novel variant oxidase enzymes are capable of oxidizing compounds towards which wild-type galactose oxidase (e.g. D-galactose: oxygen 6-oxidoreductase, GAO; EC 1.1.3.9) has little or no activity. Preferred galactose oxidase variants are those which that have improved capability to oxidize secondary alcohols and/or D-glucose relative to the wild-type enzyme.

摘要翻译： 提供了葡萄糖氧化酶，包括半乳糖氧化酶的新型变体。 编码这些新变体的多核苷酸可以在重组宿主细胞表达系统中表达。 新型变体氧化酶能够氧化化合物，野生型半乳糖氧化酶（例如D-半乳糖：氧-6-氧化还原酶，GAO; EC1.1.3.9）具有很少或没有活性。 优选的半乳糖氧化酶变体是那些相对于野生型酶具有改善氧化仲醇和/或D-葡萄糖的能力的变体。