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公开(公告)号：EP1290147A4
公开(公告)日：2004-12-08
申请号：EP00980811
申请日：2000-11-27
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.
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公开(公告)号：EP1290147A1
公开(公告)日：2003-03-12
申请号：EP00980811.4
申请日：2000-11-27
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.
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公开(公告)号：EP1228200A1
公开(公告)日：2002-08-07
申请号：EP00975458.1
申请日：2000-10-27
发明人： ARNOLD, Frances , SIEBER, Volker , ZHANG, Ji, Hu
CPC分类号： C12N15/1093 , C07K14/43595 , C12N9/0071 , C12N9/0077 , C12N15/1027
摘要： The invention relates to an improved method for creating gene and protein libraries, particularly random gene libraries encoding for hybrid proteins containing fragments from one or two parent proteins. The method may be used to make libraries of circularly permuted variants of genes encoding a single protein or hybrid proteins, especially single chain proteins. In addition, the invention can be used to make libraries for protein fragment complementation, in which the two fragments originate from one parent protein or from two different parent proteins. The method can produce a library of genes mostly of the correct size, leading to a high fraction of functional hybrids or complements. When coupled with suitable screening or selection, the method can be used to create and identify hybrid proteins, including new proteins with new or altered properties. The invention also provides libraries of hybrid proteins, especially single-chain proteins, that include an N-terminal sequence originating from one parent protein, fused to a C-terminal sequence of a second parent protein, with both sequences varying in length among the hybrids of the library.
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