公开(公告)号：US20110250674A1

公开(公告)日：2011-10-13

申请号：US13162636

申请日：2011-06-17

Applicant: Kim Vilbour Andersen ,  Martin Schülein ,  Torben Henriksen ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

Inventor： Kim Vilbour Andersen ,  Martin Schülein ,  Torben Henriksen ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

IPC: C12N9/42

CPC classification number: C11D3/38645 ,  C12N9/2437 ,  C12Y302/01004

Abstract: The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.

Abstract translation: 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。

公开(公告)号：US20050009166A1

公开(公告)日：2005-01-13

申请号：US10919195

申请日：2004-08-16

Applicant: Kim Andersen ,  Martin Schulein ,  Hanne Dela ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

Inventor： Kim Andersen ,  Martin Schulein ,  Hanne Dela ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

IPC: C12N15/09 ,  C11D3/386 ,  C12N1/15 ,  C12N1/21 ,  C12N9/42 ,  C12R1/645 ,  C12S11/00 ,  C12S99/00 ,  D06L4/40 ,  D06M16/00 ,  D21B1/02 ,  D21C5/02 ,  D21H21/24 ,  C07H21/04 ,  C12N15/74

CPC classification number: C11D3/38645 ,  C12N9/2437 ,  C12Y302/01004

Abstract: The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.

Abstract translation: 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。

公开(公告)号：US06518042B1

公开(公告)日：2003-02-11

申请号：US09512250

申请日：2000-02-24

Applicant: Torben Vedel Borchert ,  Lars Christiansen ,  Jesper Vind

Inventor： Torben Vedel Borchert ,  Lars Christiansen ,  Jesper Vind

IPC: C12P2102

CPC classification number: C07K14/38 ,  C12N15/80

Abstract: A process for making DNA libraries in filamentous fungal cells using a novel cloned gene involved in the mismatch repair system of filamentous fungal cells.

Abstract translation: 使用涉及丝状真菌细胞的错配修复系统的新的克隆基因在丝状真菌细胞中制备DNA文库的方法。

公开(公告)号：US4598048A

公开(公告)日：1986-07-01

申请号：US591461

申请日：1984-03-20

Applicant: Boerge K. Diderichsen ,  Lars Christiansen

Inventor： Boerge K. Diderichsen ,  Lars Christiansen

IPC: C12P19/22 ,  C12N9/26 ,  C12N15/09 ,  C12R1/07 ,  C12R1/125 ,  C12R1/19 ,  C12N15/00

CPC classification number: C12N9/2411 ,  C12N9/2408 ,  Y10S435/832

Abstract: A maltogenic amylase enzyme with improved thermostability, which can be produced by cultivating Bacillus strain NCIB 11837 belonging to the Bacillus stearothermophilus complex, is made by cultivation of a host microorganism transformed with the gene coding for the maltogenic amylase enzyme.

Abstract translation: 通过培养属于嗜热脂肪芽孢杆菌复合物的芽孢杆菌菌株NCIB 11837可以通过培养用编码麦芽糖淀粉酶的基因转化的宿主微生物来制备具有改善的热稳定性的麦芽糖淀粉酶。

公开(公告)号：US5726038A

公开(公告)日：1998-03-10

申请号：US446646

申请日：1995-05-25

Applicant: Lars Christiansen ,  Jens Gunner Litske Petersen

Inventor： Lars Christiansen ,  Jens Gunner Litske Petersen

IPC: C12N1/19 ,  C07K14/81 ,  C12N9/60 ,  C12N15/09 ,  C12N15/62 ,  C12N15/81 ,  C12P21/02 ,  C12R1/865 ,  C12N15/00

CPC classification number: C07K14/8103 ,  C12N15/625 ,  C12N15/81 ,  C12N9/60 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/036

Abstract: A DNA construct comprising the following sequence: 5'-P-SP-(LP).sub.n -PS-HP-3' wherein P is a promoter sequence, SP is a DNA sequence encoding the yeast aspartic protease 3 (YAP3) signal peptide, LP is a DNA sequence encoding a leader peptide, n is 0 or 1, PS is a DNA sequence encoding a peptide defining a yeast processing site, and HP is a DNA sequence encoding a polypeptide which is heterologous to a selected host organism. The YAP3 signal peptide provides efficient secretion of heterologous proteins in yeast.

Abstract translation: PCT No.PCT / DK94 / 00281 Sec。 371日期：1995年5月25日 102（e）日期1995年5月25日PCT提交1994年7月8日PCT公布。 第WO95 / 02059号公报 日期1995年1月19日包含以下序列的DNA构建体：5'-P-SP-（LP）n-PS-HP-3'，其中P是启动子序列，SP是编码酵母天冬氨酸蛋白酶3的DNA序列 YAP3）信号肽，LP是编码前导肽的DNA序列，n是0或1，PS是编码限定酵母加工位点的肽的DNA序列，HP是编码与选择的异源的多肽的DNA序列 宿主生物。 YAP3信号肽在酵母中提供异源蛋白质的高效分泌。

公开(公告)号：US08017372B2

公开(公告)日：2011-09-13

申请号：US11830063

申请日：2007-07-30

Applicant: Kim Vilbour Andersen ,  Martin Schulein ,  Torben Henriksen, legal representative ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

Inventor： Kim Vilbour Andersen ,  Martin Schulein ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

IPC: C12N9/42 ,  C12N15/00 ,  C12P21/04 ,  C07H21/04

CPC classification number: C11D3/38645 ,  C12N9/2437 ,  C12Y302/01004

Abstract: The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.

Abstract translation: 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。

公开(公告)号：US20080206836A1

公开(公告)日：2008-08-28

申请号：US11830063

申请日：2007-07-30

Applicant: Kim Vilbour Andersen ,  Martin Schulein ,  Torben Henriksen ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

Inventor： Kim Vilbour Andersen ,  Martin Schulein ,  Torben Henriksen ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

IPC: C12N9/42

CPC classification number: C11D3/38645 ,  C12N9/2437 ,  C12Y302/01004

Abstract: The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.

Abstract translation: 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。

公开(公告)号：US20050019645A1

公开(公告)日：2005-01-27

申请号：US10483388

申请日：2002-07-13

Applicant: Stephen Thomas ,  lars Christiansen ,  Yeesan Le ,  Jeremy Lawrence

Inventor： Stephen Thomas ,  lars Christiansen ,  Yeesan Le ,  Jeremy Lawrence

IPC: H01M2/00 ,  H01M2/08 ,  H01M2/14 ,  H01M2/18 ,  H01M8/0247 ,  H01M8/0271 ,  H01M8/0282 ,  H01M8/2425 ,  H01M8/12 ,  H01M8/24

CPC classification number: H01M8/0282 ,  H01M8/0247 ,  H01M8/0271 ,  H01M8/2425 ,  H01M8/2435 ,  H01M8/2483

Abstract: A fuel cell stack comprising alternating solid oxide fuel cell plates (10) and gas separator plates (30) stacked face to face with one or more seal assemblies (34, 36, 37, 38, 40) provided between opposed generally planar surfaces (33, 54) of each adjacent pair of plates. Each seal assembly comprises a pair of rigid ribs (36, 37) projecting from one surface (33) with a valley (38) therebetween and a third rigid rib (34) projecting from the other surface (54) and nested between the pair of ribs. Opposed contact surfaces (54, 60, 61) cooperate to define the maximum insertion of the third rib (34) into the valley (38). The third rib (34) has a profile that leaves a void between the valley (38) and the third rib (34) at said maximum insertion. A glass sealant (40) in said void contacts the surface of the valley (38) and the third rib (34). In a preferred embodiment each rib (34, 36, 37) tapers away from the respective surface (33, 54) towards a distal surface (51, 61) of the rib.

Abstract translation: 一种燃料电池堆，包括交替的固体氧化物燃料电池板（10）和与一个或多个密封组件（34,36,37,38,40）面对面地叠置的固体氧化物燃料电池板（30），所述密封组件设置在相对的大致平坦的表面（33 ，54）。 每个密封组件包括从一个表面（33）突出的一对刚性肋（36,37），其间具有谷部（38），以及从另一个表面（54）突出并嵌套在所述一对 肋骨。 相互接触的表面（54,60,61）协作以限定第三肋（34）进入谷部（38）的最大插入。 第三肋（34）具有在所述最大插入时在谷（38）和第三肋（34）之间留下空隙的轮廓。 所述空隙中的玻璃密封剂（40）接触谷（38）和第三肋（34）的表面。 在优选实施例中，每个肋（34,36,37）朝向肋的远侧表面（51,61）逐渐远离相应的表面（33,54）。

公开(公告)号：US20120289450A1

公开(公告)日：2012-11-15

申请号：US13471757

申请日：2012-05-15

Applicant: Kim Vilbour Andersen ,  Martin Schülein ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

Inventor： Kim Vilbour Andersen ,  Martin Schülein ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

IPC: C12N9/42 ,  C11D3/386

CPC classification number: C11D3/38645 ,  C12N9/2437 ,  C12Y302/01004

Abstract: The present invention relates to cellulase variants, i.e., endo-beta-1,4-glucanase variants, derived from a parental cellulase, i.e., endo-beta-1,4-glucanase, by substitution, insertion and/or deletion, which variant has a catalytic core domain, in which the variant at position 5 holds an alanine residue (A), a serine residue (S), or a threonine residue (T); at position 8 holds a phenylalanine residue (F), or a tyrosine residue (Y); at position 9 holds a phenylalanine residue (F), a tryptophan residue (W), or a tyrosine residue (Y); at position 10 holds an aspartic acid residue (D); and at position 121 holds an aspartic acid residue (D).

Abstract translation: 本发明涉及通过取代，插入和/或缺失衍生自亲代纤维素酶（即内切-β-1,4-葡聚糖酶）的纤维素酶变体，即内切-β-1,4-葡聚糖酶变体，该变体 具有催化核心结构域，其中第5位的变体含有丙氨酸残基（A），丝氨酸残基（S）或苏氨酸残基（T）; 在8位保持苯丙氨酸残基（F）或酪氨酸残基（Y）; 位置9含有苯丙氨酸残基（F），色氨酸残基（W）或酪氨酸残基（Y）; 在10位保持天冬氨酸残基（D）; 并且在位置121处保持天冬氨酸残基（D）。

公开(公告)号：US5316923A

公开(公告)日：1994-05-31

申请号：US973483

申请日：1992-11-09

Applicant: Lars Christiansen

Inventor： Lars Christiansen

IPC: C12N15/09 ,  C07K14/00 ,  C07K14/395 ,  C07K14/575 ,  C07K14/62 ,  C12N1/19 ,  C12N15/62 ,  C12N15/81 ,  C12P21/02 ,  C12R1/865 ,  C07K7/10

CPC classification number: C07K14/62 ,  C12N15/625 ,  C12N15/815 ,  C07K2319/036 ,  C07K2319/75

Abstract: Synthetic leaders for effective secretion of proteins in yeast are provided. Also provided are replicable yeast vectors containing a DNA-sequence encoding the synthetic leader positioned upstream to a DNA-sequence encoding the desired product and operably connected with promoter and signal sequences. There are also provided yeast strains transformed with such vectors and a method for producing proteins by means of the transformed yeast strains.

Abstract translation: 提供了在酵母中有效分泌蛋白质的合成领导者。 还提供了可复制的酵母载体，其含有编码合成前导序列的DNA序列，所述DNA序列位于编码所需产物的DNA序列的上游，并与启动子和信号序列可操作地连接。 还提供了用这种载体转化的酵母菌株和通过转化的酵母菌株产生蛋白质的方法。