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公开(公告)号：US20040171826A1
公开(公告)日：2004-09-02
申请号：US10695243
申请日：2003-10-27
发明人： Stephen Hamilton
IPC分类号： C07H021/04 , C12N001/18 , C07K014/47 , C12N015/74
CPC分类号： C12P21/005 , C12N9/1051 , C12N9/2488 , C12Y302/0113
摘要： The present invention generally relates to methods of modifying the glycosylation structures of recombinant proteins expressed in fungi or other lower eukaryotes, to more closely resemble the glycosylation of proteins from higher mammals, in particular humans. The present invention also relates to novel enzymes and, nucleic acids encoding them and, hosts engineered to express the enzymes, methods for producing modified glycoproteins in hosts and modified glycoproteins so produced.
摘要翻译： 本发明一般涉及改变在真菌或其他低等真核生物中表达的重组蛋白的糖基化结构的方法，以更加类似于来自较高哺乳动物，特别是人的蛋白质的糖基化。 本发明还涉及新的酶和编码它们的核酸，以及工程化表达酶的宿主，在宿主中产生修饰的糖蛋白的方法和如此制备的修饰的糖蛋白。
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2.发明授权公开(公告)号：US09765315B2
公开(公告)日：2017-09-19
申请号：US14421774
申请日：2013-08-15
CPC分类号： C12N9/2402 , C12N9/2408 , C12N9/2428 , C12N9/2437 , C12N9/244 , C12N9/2445 , C12N9/2471 , C12N9/2482 , C12N9/2494 , C12N9/88 , C12Y302/01037 , C12Y302/01051 , C12Y302/01055 , C12Y302/01059 , C12Y302/01075 , C12Y302/01089 , C12Y302/0113 , C12Y302/01177
摘要： The present invention provides nucleotide sequences of Macrophomina phaseolina (“M. phaseolina”) that encodes proteins/enzymes with cellulolytic activity, including a cellulase activity, a endoglucanase, a cellobiohydrolase, a β-glucosidase, a a-glucosidase, a xylanase, a mannanse, a β-xylosidase, a a-xylosidase, a galactosidase, an arabinofuranosidase, a a-fucosidases, a β-galactanase, an unsaturated β-glucuronyl hydrolase and/or oligomerase activity. Vectors, expression constructs and host cells comprising and/or consisting of the nucleotide sequences of the enzyme genes are also provided. The invention further provides methods for producing the enzymes and methods for modifying the enzymes in order to improve their desirable characteristics. The enzymes of the invention can be used in a variety of, but not limited to, pharmaceutical, agricultural, food and feed processing, biofuel, energy efficiency and industrial contexts. These enzymes are also useful for complete hydrolysis of lignocellulosic biomass into simple sugar that can then be fermented to liquid fuels and chemical feedstocks.
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公开(公告)号：US08986949B2
公开(公告)日：2015-03-24
申请号：US13070946
申请日：2011-03-24
申请人： Stephen Hamilton
发明人： Stephen Hamilton
CPC分类号： C12P21/005 , C12N9/1051 , C12N9/2488 , C12Y302/0113
摘要： The present invention generally relates to methods of modifying the glycosylation structures of recombinant proteins expressed in fungi or other lower eukaryotes, to more closely resemble the glycosylation of proteins from higher mammals, in particular humans. The present invention also relates to novel enzymes and, nucleic acids encoding them and, hosts engineered to express the enzymes, methods for producing modified glycoproteins in hosts and modified glycoproteins so produced.
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公开(公告)号：US11866715B2
公开(公告)日：2024-01-09
申请号：US17528619
申请日：2021-11-17
CPC分类号： C12N15/815 , C12N9/1051 , C12N9/2488 , C12Y204/01232 , C12Y302/0113
摘要： Disclosed herein are novel Pichia pastoris strains for expression of exogenous proteins with substantially homogeneous N-glycans. The strains are genetically engineered to include a mutant OCH1 allele which is transcribed into an mRNA coding for a mutant OCH1 gene product (i.e., α-1,6-mannosyltransferase, or “OCH1 protein”). The mutant OCH1 protein contains a catalytic domain substantially identical to that of the wild type OCH1 protein, but lacks an N-terminal sequence necessary to target the OCH1 protein to the Golgi apparatus. The strains disclosed herein are robust, stable, and transformable, and the mutant OCH1 allele and the ability to produce substantially homogeneous N-glycans are maintained for generations after rounds of freezing and thawing and after subsequent transformations.
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公开(公告)号：US09617550B2
公开(公告)日：2017-04-11
申请号：US14437683
申请日：2013-10-23
CPC分类号： C12N15/815 , C12N9/1051 , C12N9/2488 , C12Y204/01232 , C12Y302/0113
摘要： Disclosed herein are novel Pichia pastoris strains for expression of exogenous proteins with substantially homogeneous N-glycans. The strains are genetically engineered to include a mutant OCH1 allele which is transcribed into an mRNA coding for a mutant OCH1 gene product (i.e., α-1,6-mannosyltransferase, or “OCH1 protein”). The mutant OCH1 protein contains a catalytic domain substantially identical to that of the wild type OCH1 protein, but lacks an N-terminal sequence necessary to target the OCH1 protein to the Golgi apparatus. The strains disclosed herein are robust, stable, and transformable, and the mutant OCH1 allele and the ability to produce substantially homogeneous N-glycans are maintained for generations after rounds of freezing and thawing and after subsequent transformations.
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6.发明申请PICHIA PASTORIS STRAINS FOR PRODUCING PREDOMINANTLY HOMOGENEOUS GLYCAN STRUCTURE 有权PICHIA PASTORIS菌株，用于生产均质的均质结构公开(公告)号：US20150267212A1
公开(公告)日：2015-09-24
申请号：US14437683
申请日：2013-10-23
CPC分类号： C12N15/815 , C12N9/1051 , C12N9/2488 , C12Y204/01232 , C12Y302/0113
摘要： Disclosed herein are novel Pichia pastoris strains for expression of exogenous proteins with substantially homogeneous N-glycans. The strains are genetically engineered to include a mutant OCH1 allele which is transcribed into an mRNA coding for a mutant OCH1 gene product (i.e., α-1,6-mannosyltransferase, or “OCH1 protein”). The mutant OCH1 protein contains a catalytic domain substantially identical to that of the wild type OCH1 protein, but lacks an N-terminal sequence necessary to target the OCH1 protein to the Golgi apparatus. The strains disclosed herein are robust, stable, and transformable, and the mutant OCH1 allele and the ability to produce substantially homogeneous N-glycans are maintained for generations after rounds of freezing and thawing and after subsequent transformations.
摘要翻译： 本文公开了用于表达具有基本上均匀的N-聚糖的外源蛋白质的新型毕赤酵母菌株。 菌株被遗传工程化以包括转录成编码突变型OCH1基因产物（即α-1,6-甘露糖基转移酶或“OCH1蛋白”）的mRNA的突变型OCH1等位基因。 突变体OCH1蛋白含有与野生型OCH1蛋白基本相同的催化结构域，但缺少将OCH1蛋白靶向高尔基体的必需的N-末端序列。 本文公开的菌株是稳健的，稳定的和可转化的，突变体OCH1等位基因和产生基本上均一的N-聚糖的能力在几轮冷冻和融化之后以及随后的转化后保持。
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公开(公告)号：US07332299B2
公开(公告)日：2008-02-19
申请号：US10695243
申请日：2003-10-27
申请人： Stephen Hamilton
发明人： Stephen Hamilton
CPC分类号： C12P21/005 , C12N9/1051 , C12N9/2488 , C12Y302/0113
摘要： The present invention generally relates to methods of modifying the glycosylation structures of recombinant proteins expressed in fungi or other lower eukaryotes, to more closely resemble the glycosylation of proteins from higher mammals, in particular humans. The present invention also relates to novel enzymes and, nucleic acids encoding them and, hosts engineered to express the enzymes, methods for producing modified glycoproteins in hosts and modified glycoproteins so produced.
摘要翻译： 本发明一般涉及改变在真菌或其他低等真核生物中表达的重组蛋白的糖基化结构的方法，以更加类似于来自较高哺乳动物，特别是人的蛋白质的糖基化。 本发明还涉及新的酶和编码它们的核酸，以及工程化表达酶的宿主，在宿主中产生修饰的糖蛋白的方法和如此制备的修饰的糖蛋白。
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公开(公告)号：US20240352472A1
公开(公告)日：2024-10-24
申请号：US18520960
申请日：2023-11-28
CPC分类号： C12N15/815 , C12N9/1051 , C12N9/2488 , C12Y204/01232 , C12Y302/0113
摘要： Disclosed herein are novel Pichia pastoris strains for expression of exogenous proteins with substantially homogeneous N-glycans. The strains are genetically engineered to include a mutant OCH1 allele which is transcribed into an mRNA coding for a mutant OCH1 gene product (i.e., α-1,6-mannosyltransferase, or “OCH1 protein”). The mutant OCH1 protein contains a catalytic domain substantially identical to that of the wild type OCH1 protein, but lacks an N-terminal sequence necessary to target the OCH1 protein to the Golgi apparatus. The strains disclosed herein are robust, stable, and transformable, and the mutant OCH1 allele and the ability to produce substantially homogeneous N-glycans are maintained for generations after rounds of freezing and thawing and after subsequent transformations.
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公开(公告)号：US20170166910A1
公开(公告)日：2017-06-15
申请号：US15444870
申请日：2017-02-28
CPC分类号： C12N15/815 , C12N9/1051 , C12N9/2488 , C12Y204/01232 , C12Y302/0113
摘要： Disclosed herein are novel Pichia pastoris strains for expression of exogenous proteins with substantially homogeneous N-glycans. The strains are genetically engineered to include a mutant OCH1 allele which is transcribed into an mRNA coding for a mutant OCH1 gene product (i.e., α-1,6-mannosyltransferase, or “OCH1 protein”). The mutant OCH1protein contains a catalytic domain substantially identical to that of the wild type OCH1 protein, but lacks an N-terminal sequence necessary to target the OCH1 protein to the Golgi apparatus. The strains disclosed herein are robust, stable, and transformable, and the mutant OCH1 allele and the ability to produce substantially homogeneous N-glycans are maintained for generations after rounds of freezing and thawing and after subsequent transformations.
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10.发明申请CELLULOSE AND/OR HEMICELLULOSES DEGRADING ENZYMES FROM MACROPHOMINA PHASEOLINA AND USES THEREOF 有权纤维素和/或致细胞毒素降解大蒜素的酶及其用途公开(公告)号：US20150291945A1
公开(公告)日：2015-10-15
申请号：US14421774
申请日：2013-08-15
CPC分类号： C12N9/2402 , C12N9/2408 , C12N9/2428 , C12N9/2437 , C12N9/244 , C12N9/2445 , C12N9/2471 , C12N9/2482 , C12N9/2494 , C12N9/88 , C12Y302/01037 , C12Y302/01051 , C12Y302/01055 , C12Y302/01059 , C12Y302/01075 , C12Y302/01089 , C12Y302/0113 , C12Y302/01177
摘要： The present invention provides nucleotide sequences of Macrophomina phaseolina (“M. phaseolina”) that encodes proteins/enzymes with cellulolytic activity, including a cellulase activity, a endoglucanase, a cellobiohydrolase, a β-glucosidase, a a-glucosidase, a xylanase, a mannanse, a β-xylosidase, a a-xylosidase, a galactosidase, an arabinofuranosidase, a a-fucosidases, a β-galactanase, an unsaturated β-glucuronyl hydrolase and/or oligomerase activity. Vectors, expression constructs and host cells comprising and/or consisting of the nucleotide sequences of the enzyme genes are also provided. The invention further provides methods for producing the enzymes and methods for modifying the enzymes in order to improve their desirable characteristics. The enzymes of the invention can be used in a variety of, but not limited to, pharmaceutical, agricultural, food and feed processing, biofuel, energy efficiency and industrial contexts. These enzymes are also useful for complete hydrolysis of lignocellulosic biomass into simple sugar that can then be fermented to liquid fuels and chemical feedstocks.
摘要翻译： 本发明提供了编码具有纤维素分解活性的蛋白质/酶的Macrophomina phaseolina（“M. phaseolina”）的核苷酸序列，包括纤维素酶活性，内切葡聚糖酶，纤维二糖水解酶，葡糖苷酶，α-葡糖苷酶，木聚糖酶， 甘露聚糖，α-木糖苷酶，α-木糖苷酶，半乳糖苷酶，阿拉伯呋喃糖苷酶，α-岩藻糖苷酶，β-半乳糖苷酶，不饱和的 - 葡萄糖醛酸水解酶和/或寡聚酶活性。 也提供了包含和/或由该酶基因的核苷酸序列组成的载体，表达构建体和宿主细胞。 本发明还提供了用于生产酶的方法和用于改变酶的方法，以便改善其期望的特性。 本发明的酶可以多种但不限于制药，农业，食品和饲料加工，生物燃料，能源效率和工业环境。 这些酶也可用于将木质纤维素生物质完全水解成单糖，然后将其发酵成液体燃料和化学原料。
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