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1. 发明授权

US5866408A Modification of xylanase to improve thermophilicity, alkophilicity and thermostability 失效
标题翻译：修饰木聚糖酶以改善嗜热性，亲油性和热稳定性
公开(公告)号：US5866408A
公开(公告)日：1999-02-02
申请号：US47370
申请日：1998-03-25
申请人： Wing L. Sung , Makoto Yaguchi , Kazuhiko Ishikawa
发明人： Wing L. Sung , Makoto Yaguchi , Kazuhiko Ishikawa
IPC分类号： C12N15/09 , C12N9/24 , C12N15/00 , C12N15/56 , C12R1/09 , C12R1/885 , D21C5/00 , D21C9/10 , D21C3/00
CPC分类号： C12Y302/01032 , C12N9/248 , C12Y302/01008 , D21C5/005 , C07K2319/00
摘要： Producing a xylanase enzyme of superior performance in the bleaching of pulp. More specifically, a modified xylanase of Family 11 that shows improved thermophilicity, alkalophilicity, and thermostability as compared to the natural xylanase. The modified xylanases contain any of three types of modifications: (1) changing amino acids 10, 27, and 29 of Trichoderma reesei xylanase II or the corresponding amino acids of another Family 11 xylanase, where these amino acids are changed to histidine, methionine, and leucine, respectively; (2) substitution of amino acids in the N-terminal region with amino acids from another xylanase enzyme. In a preferred embodiment, substitution of the natural Bacillus circulans or Trichoderma reesei xylanase with a short sequence of amino acids from Thermomonospora fusca xylanase yielded chimeric xylanases with higher thermophilicity and alkalophilicity; (3) an extension upstream of the N-terminus of up to 10 amino acids. In a preferred embodiment, extension of the N-terminus of the xylanase with the tripeptide glycine-arginine-arginine improved its performance.
摘要翻译：在木浆漂白中生产出优异的木聚糖酶。更具体地，与天然木聚糖酶相比，家族11的改性木聚糖酶显示出改善的嗜热性，嗜碱性和热稳定性。修饰的木聚糖酶包含三种类型的修饰中的任一种：（1）改变里氏木霉木聚糖酶II的氨基酸10,27和29或另一种家族11木聚糖酶的相应氨基酸，其中这些氨基酸被改变为组氨酸，甲硫氨酸，和亮氨酸; （2）用来自另一种木聚糖酶的氨基酸取代N-末端区域中的氨基酸。在一个优选的实施方案中，用热单孢菌木聚糖酶短链氨基酸取代天然环状芽孢杆菌或里氏木霉木聚糖酶得到具有较高嗜热性和嗜碱性的嵌合木聚糖酶; （3）N-末端上游至多10个氨基酸的延伸。在优选的实施方案中，木聚糖酶的N-末端与三肽甘氨酸 - 精氨酸 - 精氨酸的延伸改善了其性能。

2. 发明授权

US5759840A Modification of xylanase to improve thermophilicity, alkalophilicity and thermostability 失效
标题翻译：改良木聚糖酶以改善嗜热性，嗜碱性和热稳定性
公开(公告)号：US5759840A
公开(公告)日：1998-06-02
申请号：US709912
申请日：1996-09-09
申请人： Wing L. Sung , Makoto Yaguchi , Kazuhiko Ishikawa
发明人： Wing L. Sung , Makoto Yaguchi , Kazuhiko Ishikawa
IPC分类号： C12N15/09 , C12N9/24 , C12N15/00 , C12N15/56 , C12R1/09 , C12R1/885 , D21C5/00 , D21C9/10
CPC分类号： C12Y302/01032 , C12N9/248 , C12Y302/01008 , D21C5/005 , C07K2319/00
摘要： Producing a xylanase enzyme of superior performance in the bleaching of pulp. More specifically, a modified xylanase of Family 11 that shows improved thermophilicity, alkalophilicity, and thermostability as compared to the natural xylanase. The modified xylanases contain any of three types of modifications: (1) changing amino acids 10, 27, and 29 of Trichoderma reesei xylanase II or the corresponding amino acids of another Family 11 xylanase, where these amino acids are changed to histidine, methionine, and leucine, respectively; (2) substitution of amino acids in the N-terminal region with amino acids from another xylanase enzyme. In a preferred embodiment, substitution of the natural Bacillus circulans or Trichoderma reesei xylanase with a short sequence of amino acids from Thermomonospora fusca xylanase yielded chimeric xylanases with higher thermophilicity and alkalophilicity; (3) an extension upstream of the N-terminus of up to 10 amino acids. In a preferred embodiment, extension of the N-terminus of the xylanase with the tripeptide glycine-arginine-arginine improved its performance.
摘要翻译：在木浆漂白中生产出优异的木聚糖酶。更具体地，与天然木聚糖酶相比，家族11的改性木聚糖酶显示出改善的嗜热性，嗜碱性和热稳定性。修饰的木聚糖酶包含三种类型的修饰中的任何一种：（1）改变里氏木霉木聚糖酶II的氨基酸10,27和29或另一种家族11木聚糖酶的相应氨基酸，其中这些氨基酸被改变为组氨酸，甲硫氨酸，和亮氨酸; （2）用来自另一种木聚糖酶的氨基酸取代N-末端区域中的氨基酸。在一个优选的实施方案中，用热单孢菌木聚糖酶短链氨基酸取代天然环状芽孢杆菌或里氏木霉木聚糖酶得到具有较高嗜热性和嗜碱性的嵌合木聚糖酶; （3）N-末端上游至多10个氨基酸的延伸。在优选的实施方案中，木聚糖酶的N-末端与三肽甘氨酸 - 精氨酸 - 精氨酸的延伸改善了其性能。

3. 发明授权

US5405769A Construction of thermostable mutants of a low molecular mass xylanase 失效
标题翻译：构建低分子量木聚糖酶的耐热突变体
公开(公告)号：US5405769A
公开(公告)日：1995-04-11
申请号：US44621
申请日：1993-04-08
申请人： Robert L. Campbell , David R. Rose , Wing L. Sung , Makoto Yaguchi , Warren W. Wakarchuk
发明人： Robert L. Campbell , David R. Rose , Wing L. Sung , Makoto Yaguchi , Warren W. Wakarchuk
IPC分类号： C12N9/24 , D21C5/00
CPC分类号： C12Y302/01032 , C12N9/248 , C12Y302/01008 , D21C5/005
摘要： The thermostability of the 20,396 dalton Bacillus circulans xylanase was increased by site-directed mutagenesis. The thermostability was conferred by the presence of non-native disulfide bridges, and selected N-terminal mutations. The introduction of these non-native disulfide bridges was accomplished by the examination of the three-dimensional structure of the enzyme, and choosing sites where a favorable geometry for a bridge existed. The N-terminal mutations were constructed on the basis of primary sequence comparison with other family G xylanases. The mutant proteins were examined for their ability to retain enzymatic activity after heating as an indication of increased thermostability. These thermotolerant variants are useful as an alternative to chemical bleaching of Kraft pulp in a pre-bleaching step (bio-bleaching). The pre-bleaching involves temperatures higher than that normally used for these enzymes and accordingly these thermotolerant variants can be advantageously used at this step. Thermotolerant xylanases are also of use in the food processing industry.
摘要翻译：通过定点诱变增加了20396道尔顿环状芽孢杆菌木聚糖酶的热稳定性。耐热性由非天然二硫键的存在和选择的N端突变赋予。这些非天然二硫键的引入是通过检查酶的三维结构，并选择存在桥的有利几何形状的位点来实现的。基于与其他家族G木聚糖酶的初级序列比较构建N末端突变。检查突变体蛋白质在加热后保留酶活性的能力，作为增加热稳定性的指标。这些耐热变体在预漂白步骤（生物漂白）中可用作牛皮纸浆的化学漂白的替代物。预漂白涉及比通常用于这些酶的温度更高的温度，因此这些耐热变体可有利地用于该步骤。耐热木聚糖酶也可用于食品加工业。

4. 发明授权

US07510860B1 Xylanases with enhanced thermophilicity and alkalophilicity 有权
标题翻译：具有增强的嗜热性和嗜碱性的木聚糖酶
公开(公告)号：US07510860B1
公开(公告)日：2009-03-31
申请号：US09990874
申请日：2001-11-21
申请人： Wing L. Sung
发明人： Wing L. Sung
IPC分类号： C12N9/26 , C12P21/06 , D21C3/00 , C07H21/04
CPC分类号： C12N9/2482 , C12Y302/01008 , D21C5/005 , D21C9/1036
摘要： The present invention provides a xylanase, or a modified xylanase enzyme comprising at least one substituted amino acid residue at a position selected from the group consisting of amino acid 11, 116, 118, 144 and 161, the position determined from sequence alignment of the modified xylanase with Trichoderma reesei xylanase II amino acid sequence. The xylanases described herein exhibit improved thermophilicity, alkalophilicity, expression efficiency, or a combination thereof, in comparison to a corresponding native xylanase.
摘要翻译：本发明提供一种木聚糖酶或修饰的木聚糖酶，其在选自氨基酸11,116,118,144和161的位置包含至少一个取代的氨基酸残基，所述位置由修饰的木聚糖酶与里氏木霉木聚糖酶II氨基酸序列。与相应的天然木聚糖酶相比，本文所述的木聚糖酶表现出改善的嗜热性，嗜碱性，表达效率或其组合。

5. 发明申请

US20090075330A1 XYLANASES WITH ENHANCED THERMOPHILICITY AND ALKALOPHILICITY 有权
标题翻译：具有增强热性和碱性的XYLANASES
公开(公告)号：US20090075330A1
公开(公告)日：2009-03-19
申请号：US09990874
申请日：2001-11-21
申请人： Wing L. Sung
发明人： Wing L. Sung
IPC分类号： C12N9/24 , C12P21/06 , C07H21/04 , C12N1/18
CPC分类号： C12N9/2482 , C12Y302/01008 , D21C5/005 , D21C9/1036
摘要： The present invention provides a xylanase, or a modified xylanase enzyme comprising at least one substituted amino acid residue at a position selected from the group consisting of amino acid 11, 116, 118, 144 and 161, the position determined from sequence alignment of the modified xylanase with Trichoderma reesei xylanase II amino acid sequence. The xylanases described herein exhibit improved thermophilicity, alkalophilicity, expression efficiency, or a combination thereof, in comparison to a corresponding native xylanase.
摘要翻译：本发明提供一种木聚糖酶或修饰的木聚糖酶，其在选自氨基酸11,116,118,144和161的位置包含至少一个取代的氨基酸残基，所述位置由修饰的木聚糖酶与里氏木霉木聚糖酶II氨基酸序列。与相应的天然木聚糖酶相比，本文所述的木聚糖酶表现出改善的嗜热性，嗜碱性，表达效率或其组合。

6. 发明授权

US08927248B2 Modification of xylanases to increase thermophilicity, thermostability and alkalophilicity 有权
标题翻译：改良木聚糖酶以增加嗜热性，热稳定性和嗜碱性
公开(公告)号：US08927248B2
公开(公告)日：2015-01-06
申请号：US12253774
申请日：2008-10-17
申请人： Wing L. Sung
发明人： Wing L. Sung
IPC分类号： C12N9/24 , D21H17/00 , D21C5/00 , D21C9/10
CPC分类号： C12N9/2482 , C12Y302/01008 , D21C5/005 , D21C9/1036 , D21H17/005
摘要： A modified Family 11 xylanase enzyme comprising cysteine residues at positions 99 and 118 to form an intramolecular disulfide bond is provided. The modified xylanase is produced by substitution of an amino acid at position 99, 118 or both positions 99 and 118 with a cysteine to produce the intramolecular disulfide bond. Xylanases of the invention display improved thermophilicity, alkalophilicity or thermostability relative to wild-type xylanases. Such xylanases find use in a variety of applications in industry that require enzyme activities at temperatures and/or pH values above that of the native enzyme.
摘要翻译：提供了修饰的家族11木聚糖酶，其包含位置99和118处的半胱氨酸残基以形成分子内二硫键。经修饰的木聚糖酶是通过用半胱氨酸取代位置99,118或位置99和118的氨基酸产生分子内二硫键而产生的。本发明的木聚糖酶相对于野生型木聚糖酶显示出改善的嗜热性，嗜碱性或热稳定性。这种木聚糖酶可用于工业中需要酶活性的各种应用，温度和/或pH值高于天然酶的温度和/或pH值。

7. 发明申请

US20110312066A1 ENZYMATIC PREPARATION OF PLANT FIBERS 有权
标题翻译：植物纤维的酶化制备
公开(公告)号：US20110312066A1
公开(公告)日：2011-12-22
申请号：US13144470
申请日：2009-12-23
申请人： Wing L. Sung , Mark Wood , Fang Huang
发明人： Wing L. Sung , Mark Wood , Fang Huang
IPC分类号： D21C5/00
CPC分类号： D01C1/02
摘要： A method of extracting fibers from decorticated plant bast skin involves pre-treating decorticated plant bast skin of a fiber plant with an aqueous solution containing trisodium citrate having a pH in a range of about 8-14 at a temperature of about 90° C. or less; and subsequently treating recovered fibers with a protease at alkaline pH.
摘要翻译：从脱皮的植物皮肤皮肤提取纤维的方法包括用约含有柠檬酸三钠的水溶液预处理纤维植物的脱皮植物皮肤，所述柠檬酸三钠在约90℃的温度下约为8-14℃，或减; 并随后在碱性pH下用蛋白酶处理回收的纤维。

8. 发明授权

US08591701B2 Extraction of hemp fibers 有权
标题翻译：大麻纤维的提取
公开(公告)号：US08591701B2
公开(公告)日：2013-11-26
申请号：US12303931
申请日：2007-05-14
申请人： Wing L. Sung , Mark Wood , Fang Huang
发明人： Wing L. Sung , Mark Wood , Fang Huang
IPC分类号： D21C7/14
CPC分类号： D01C1/02
摘要： A method of extracting hemp fibers from decorticated hemp bast skin involves pre-treating the decorticated hemp bast skin with an aqueous solution containing di-sodium citrate, tri-sodium citrate or a mixture thereof having a pH of from about 6-13 at temperature of about 90° C. or less; and subsequently treating recovered fiber with a enzyme. Determining the extent of completion of a plant fiber degumming process involves treating degummed fiber with a recombinant pectinase expressed in an organism that produces neither cellulose nor xylanase, to release reducing sugar from any residua pectin on the degummed fiber, and, quantifying the released reducing sugar.
摘要翻译：从去皮大麻皮肤皮肤中提取大麻纤维的方法包括用含有柠檬酸二钠，柠檬酸三钠或其混合物的水溶液预处理脱皮的大麻皮肤，所述水溶液的pH为约6-13，温度为约90℃或更低; 然后用酶处理回收的纤维。确定植物纤维脱胶方法的完成程度包括用生物体中表达的重组果胶酶处理脱胶纤维，其既不产生纤维素也不生成木聚糖酶，从脱胶纤维上的任何残余果胶中释放还原糖，并定量释放的还原糖。

9. 发明申请

US20090325267A1 Modified xylanases exhibiting increased thermophilicity and alkalophilicity 失效
标题翻译：表现出增加嗜热性和嗜碱性的改性木聚糖酶
公开(公告)号：US20090325267A1
公开(公告)日：2009-12-31
申请号：US11377644
申请日：2006-03-17
申请人： Wing L. Sung
发明人： Wing L. Sung
IPC分类号： C12N9/24
CPC分类号： C12N9/2482 , C12Y302/01008
摘要： The present invention pertains to modified xylanase enzymes that exhibit increased thermostability and alkalophilicity, when compared with their native counterparts. Several modified xylanases exhibiting these properties are disclosed including xylanases with at least one modification at amino acid position 10, 27, 29, 75, 104, 105, 125, 129, 132, 135, 144, 157, 161, 162 or 165, or a combination thereof. Also included within the present invention is a modified xylanase that comprises at least one substituted amino acid residue and that may be characterized as having a maximum effective temperature (MET) between about 69° C. and about 78° C., wherein the modified xylanase is a Family 11 xylanase obtained from a Trichoderma sp. The present invention also includes a modified Family 11 xylanase obtained from a Trichoderma sp. characterized as having a maximum effective pH (MEP) between 5.8 and about 7.6. Modified xylanases characterized as having a MET between about 69° C. and about 78° C. and a MET between about 5.8 and 7.6 are also disclosed.
摘要翻译：本发明涉及与其天然对应物相比显示增加的热稳定性和嗜碱性的改性木聚糖酶。公开了显示这些性质的几种改性的木聚糖酶，其包括在氨基酸位置10,27,29，75，104，105，125，129，132，135，144，157，161，162或165具有至少一个修饰的木聚糖酶，或其组合。本发明还包括修饰的木聚糖酶，其包含至少一个取代的氨基酸残基并且可以被表征为在约69℃至约78℃之间具有最大有效温度（MET），其中所述改性木聚糖酶是从木霉属种获得的家族11木聚糖酶。本发明还包括由木霉属（Trichoderma sp。）获得的经修饰的家族11木聚糖酶。其特征在于具有5.8至约7.6之间的最大有效pH（MEP）。还公开了特征在于具有在约69℃和约78℃之间的MET和约5.8和7.6之间的MET的改性的木聚糖酶。

10. 发明授权

US08603802B2 Enzymatic preparation of plant fibers 有权
标题翻译：植物纤维的酶制剂
公开(公告)号：US08603802B2
公开(公告)日：2013-12-10
申请号：US13144470
申请日：2009-12-23
申请人： Wing L. Sung , Mark Wood , Fang Huang
发明人： Wing L. Sung , Mark Wood , Fang Huang
IPC分类号： D21C1/00 , D01C1/04 , C12Q1/37
CPC分类号： D01C1/02
摘要： A method of extracting fibers from decorticated plant bast skin involves pre-treating decorticated plant bast skin of a fiber plant with an aqueous solution containing trisodium citrate having a pH in a range of about 8-14 at a temperature of about 90° C. or less; and subsequently treating recovered fibers with a protease at alkaline pH.
摘要翻译：从脱皮的植物皮肤皮肤提取纤维的方法包括用约含有柠檬酸三钠的水溶液预处理纤维植物的脱皮植物皮肤，所述柠檬酸三钠在约90℃的温度下约为8-14℃，或减; 并随后在碱性pH下用蛋白酶处理回收的纤维。

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