公开(公告)号：US07375197B2

公开(公告)日：2008-05-20

申请号：US10031496

申请日：2002-01-14

申请人： William S. Adney ,  Stephen R. Decker ,  Suzanne Mc Carter ,  John O. Baker ,  Raphael Nieves ,  Michael E. Himmel ,  Todd B. Vinzant

发明人： William S. Adney ,  Stephen R. Decker ,  Suzanne Mc Carter ,  John O. Baker ,  Raphael Nieves ,  Michael E. Himmel ,  Todd B. Vinzant

IPC分类号： C07H21/04

CPC分类号： C12Y302/01091 ,  C12N9/2437

摘要： The disclosure provides a method for preparing an active exoglucanase in a heterologous host of eukaryotic origin. The method includes mutagenesis to reduce glycosylation of the exoglucanase when expressed in a heterologous host. It is further disclosed a method to produce variant cellobiohydrolase that is stable at high temperature through mutagenesis.

摘要翻译： 本公开提供了在真核来源的异源宿主中制备活性外切葡聚糖酶的方法。 该方法包括当在异源宿主中表达时，用于减少外切葡聚糖酶的糖基化的突变。 还公开了通过诱变在高温下稳定的变体纤维二糖水解酶的方法。

公开(公告)号：US20090081762A1

公开(公告)日：2009-03-26

申请号：US12247594

申请日：2008-10-08

申请人： William S. Adney ,  John O. Baker ,  Stephen R. Decker ,  Yat-Chen Chou ,  Michael E. Himmel ,  Shi-You Ding

发明人： William S. Adney ,  John O. Baker ,  Stephen R. Decker ,  Yat-Chen Chou ,  Michael E. Himmel ,  Shi-You Ding

IPC分类号： C12N9/42

CPC分类号： C12N9/2437 ,  C12Y302/01004 ,  C12Y302/01091

摘要： Purified cellobiohydrolase I (glycosyl hydrolase family 7 (Cel7A)) enzymes from Penicillium funiculosum demonstrate a high level of specific performance in comparison to other Cel7 family member enzymes when formulated with purified EIcd endoglucanase from A. cellulolyticus and tested on pretreated corn stover. This result is true of the purified native enzyme, as well as recombinantly expressed enzyme, for example, that enzyme expressed in a non-native Aspergillus host. In a specific example, the specific performance of the formulation using purified recombinant Cel7A from Penicillium funiculosum expressed in A. awamori is increased by more than 200% when compared to a formulation using purified Cel7A from Trichoderma reesei.

摘要翻译： 与其他Cel7家族成员酶相比，来自青霉菌（Penicillium funiculosum）的纯化纤维二糖水解酶I（糖基水解酶家族7（Cel7A））酶表现出高水平的特异性，当用来自解纤维解卷菌的纯化的EIcd内切葡聚糖酶配制并在预处理的玉米秸秆上进行测试时。 纯化的天然酶以及重组表达的酶，例如在非天然曲霉属宿主中表达的酶的结果是真实的。 在具体实例中，与使用来自里氏木霉的纯化Cel7A的制剂相比，使用在泡盛曲霉中表达的来自青霉菌的纯化重组Cel7A的制剂的具体表现增加了超过200％。

公开(公告)号：US08283150B2

公开(公告)日：2012-10-09

申请号：US12247594

申请日：2008-10-08

申请人： William S. Adney ,  John O. Baker ,  Stephen R. Decker ,  Yat-Chen Chou ,  Michael E. Himmel ,  Shi-You Ding

发明人： William S. Adney ,  John O. Baker ,  Stephen R. Decker ,  Yat-Chen Chou ,  Michael E. Himmel ,  Shi-You Ding

IPC分类号： C07K1/00 ,  C07K14/00 ,  C07K17/00 ,  C12N9/42 ,  A23L1/202

CPC分类号： C12N9/2437 ,  C12Y302/01004 ,  C12Y302/01091

摘要： Purified cellobiohydrolase I (glycosyl hydrolase family 7 (Cel7A)) enzymes from Penicillium funiculosum demonstrate a high level of specific performance in comparison to other Cel7 family member enzymes when formulated with purified EIcd endoglucanase from A. cellulolyticus and tested on pretreated corn stover. This result is true of the purified native enzyme, as well as recombinantly expressed enzyme, for example, that enzyme expressed in a non-native Aspergillus host. In a specific example, the specific performance of the formulation using purified recombinant Cel7A from Penicillium funiculosum expressed in A. awamori is increased by more than 200% when compared to a formulation using purified Cel7A from Trichoderma reesei.

摘要翻译： 与其他Cel7家族成员酶相比，来自青霉菌（Penicillium funiculosum）的纯化纤维二糖水解酶I（糖基水解酶家族7（Cel7A））酶表现出高水平的特异性，当用来自解纤维解卷菌的纯化的EIcd内切葡聚糖酶配制并在预处理的玉米秸秆上进行测试时。 纯化的天然酶以及重组表达的酶，例如在非天然曲霉属宿主中表达的酶的结果是真实的。 在具体实例中，与使用来自里氏木霉的纯化Cel7A的制剂相比，使用在泡盛曲霉中表达的来自青霉菌的纯化重组Cel7A的制剂的具体表现增加了超过200％。

公开(公告)号：US07449550B2

公开(公告)日：2008-11-11

申请号：US10557589

申请日：2003-02-27

申请人： William S. Adney ,  John O. Baker ,  Stephen R. Decker ,  Yat-Chen Chou ,  Michael E. Himmel ,  Shi-You Ding

发明人： William S. Adney ,  John O. Baker ,  Stephen R. Decker ,  Yat-Chen Chou ,  Michael E. Himmel ,  Shi-You Ding

IPC分类号： C07K14/00

CPC分类号： C12N9/2437 ,  C12Y302/01004 ,  C12Y302/01091

摘要： Purified cellobiohydrolase I (glycosyl hydrolase family 7 (Cel7A) enzymes from Penicillium funiculosum demonstrate a high level of specific performance in comparison to other Cel7 family member enzymes when formulated with purified EIcd endoglucanase from A. cellulolyticus and tested on pretreated corn stover. This result is true of the purified native enzyme, as well as recombinantly expressed enzyme, for example, that enzyme expressed in a non-native Aspergillus host. In a specific example, the specific performance of the formulation using purified recombinant Cel7A from Penicillium funiculosum expressed in A. awamori is increased by more than 200% when compared to a formulation using purified Cel7A from Trichoderma reesei.

摘要翻译： 与其他Cel7家族成员酶相比，纯化的纤维二糖水解酶I（糖基水解酶家族7（Cel7A））来自A. cellulolyticus的纯化EIcd内切葡聚糖酶，并与预处理的玉米秸秆进行了测试，结果表明： 纯化的天然酶，以及重组表达的酶，例如在非天然曲霉属宿主中表达的酶。在一个具体实例中，使用在A.A中表达的来自Penicillium funiculosum的纯化重组Cel7A的制剂的具体性能。 与使用来自里氏木霉的纯化Cel7A的制剂相比，泡盛曲增加超过200％。

公开(公告)号：US5712142A

公开(公告)日：1998-01-27

申请号：US604913

申请日：1996-02-22

申请人： William S. Adney ,  Steven R. Thomas ,  John O. Baker ,  Michael E. Himmel ,  Yat-Chen Chou

发明人： William S. Adney ,  Steven R. Thomas ,  John O. Baker ,  Michael E. Himmel ,  Yat-Chen Chou

IPC分类号： C12N9/42 ,  C12N1/20 ,  C12P21/06 ,  C17H21/04

CPC分类号： C12P7/10 ,  C12N9/2437 ,  C12Y302/01004 ,  C12Y302/01021 ,  C07K2319/02 ,  Y02E50/16

摘要： The gene encoding Acidothermus cellulolyticus E1 endoglucanase is cloned and expressed in Pichia pastoris. A new modified E1 endoglucanase enzyme comprising the catalytic domain of the full size E1 enzyme demonstrates enhanced thermostability and is produced by two methods. The first method of producing the new modified E1 is proteolytic cleavage to remove the cellulose binding domain and linker peptide of the full size E1. The second method of producing the new modified E1 is genetic truncation of the gene encoding the full size E1 so that the catalytic domain is expressed in the expression product.

摘要翻译： 编码酸热解纤维素分解酶E1内切葡聚糖酶的基因在巴斯德毕赤酵母中克隆并表达。 包含全尺寸E1酶的催化结构域的新的修饰的E1内切葡聚糖酶表现出增强的热稳定性，并通过两种方法产生。 生产新的修饰的E1的第一种方法是蛋白水解切割以去除全尺寸E1的纤维素结合结构域和接头肽。 产生新修饰的E1的第二种方法是编码全长E1的基因的遗传截短，使得催化结构域在表达产物中表达。