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公开(公告)号:US06734344B2
公开(公告)日:2004-05-11
申请号:US09733643
申请日:2000-12-08
申请人: André J. Laroche , Timothy Yikai Huang , Michele M. Frick , Zhen-Xiang Lu , Hung Chang Huang , Kuo Joan Cheng
发明人: André J. Laroche , Timothy Yikai Huang , Michele M. Frick , Zhen-Xiang Lu , Hung Chang Huang , Kuo Joan Cheng
IPC分类号: C12N1556
CPC分类号: C12Y302/01058 , C12N9/2402
摘要: The invention provides the nucleotide sequence of a novel &bgr;-(1,3) exoglucanase gene denoted as cbeg1 of the soil-borne fungus Coniothyrium minitans. The deduced amino acid sequence of the encoded &bgr;-(1,3) exoglucanase enzyme, denoted Cbeg1, is also provided. Encoded &bgr;-(1,3) exoglucanase Cbeg1 is specific for the substrate laminarin, in that results showed no activity with other substrates tested, such as carboxymethylcellulose, barley &bgr;-glucan, lichenan, oat spelt xylan and birchwood xylan. The pH and temperature optima for &bgr;-(1,3) exoglucanase Cbeg1 are 6.0 and 57° C., respectively. Cbeg1 contains 784 amino acids, and has a predicted isoelectric point (pI) of 6.0 and molecular weight of 83,646 Daltons. The invention further provides vectors and cells comprising a nucleic acid molecule encoding the cbeg1 gene, and methods for producing &bgr;-(1,3) exoglucanase Cbeg1. The cbeg1 gene is compatible with a eukaryotic heterologous expression system, making it particularly useful for a wide range of industrial applications, such as improvement of plant resistance to fungal phytopathogens or use in ruminant microbial transgenic strategies to improve feed digestion and nutritive carbohydrate availability from forage feed. In addition, the high activity of Cbeg1 over broad pH and temperature ranges may be beneficial for use in high temperature industrial applications, such as bleaching of pulp, which require temperatures greater than 37° C. Further, Cbeg1 may complement degradation initiated by endoglucanases which release oligoglucans, in that &bgr;-(1,3) exoglucanase sequentially hydrolyzes &bgr;-(1,3) glucan fragments and is required to hydrolyze oligoglucan fragments completely to obtain D-glucose, which can be assimilated.
摘要翻译: 本发明提供了一种新颖的β-(1,3)外切葡聚糖酶基因的核苷酸序列,该基因被称为土壤真菌小圆锥菌的cbeg1。 还提供了Cbeg1编码的β-(1,3)外切葡聚糖酶的推导氨基酸序列。 编码β-(1,3)外切葡聚糖酶Cbeg1是特异性的底物昆布,结果显示没有活动与其他基底测试,如羧甲基纤维素,大麦β-葡聚糖,地衣甘蔗,燕麦拼写木聚糖和桦木木聚糖。 β-(1,3)外切葡聚糖酶Cbeg1的pH和温度分别为6.0和57℃。 Cbeg1含有784个氨基酸,预测的等电点(pI)为6.0,分子量为83,646道尔顿。 本发明还提供了包含编码cbeg1基因的核酸分子的载体和细胞,以及用于产生β-(1,3)外切葡聚糖酶Cbeg1的方法。 cbeg1基因与真核异源表达系统相容,使其特别适用于广泛的工业应用,例如改善植物对真菌植物病原体的抗性或用于反刍动物微生物转基因策略,以改善饲料消化和营养碳水化合物从饲料中的供应 饲料。 此外,Cbeg1在宽pH和温度范围内的高活性可能有利于高温工业应用,例如需要温度大于37℃的纸浆漂白,此外,Cbeg1可补充由内切葡聚糖酶引发的降解, 释放寡葡聚糖,其中β-(1,3)外切葡聚糖酶依次水解β-(1,3)葡聚糖片段,并且需要完全水解寡葡聚糖片段以获得可被同化的D-葡萄糖。
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公开(公告)号:US06121034A
公开(公告)日:2000-09-19
申请号:US311170
申请日:1999-05-13
申请人: Andre J. Laroche , Timothy Y. Huang , Michele M. Frick , Zhen-Xiang Lu , Hung Chang Huang , Kuo Joan Cheng
发明人: Andre J. Laroche , Timothy Y. Huang , Michele M. Frick , Zhen-Xiang Lu , Hung Chang Huang , Kuo Joan Cheng
CPC分类号: C12Y302/01008 , C12N9/248 , C07K2319/00
摘要: The nucleotide sequence of a novel xylanase gene obtained from the soilborne fungus Coniothyrium minitans, and denoted cxy1, is provided. The amino acid sequence of the encoded xylanase enzyme, denoted Cxy1, is also provided.
摘要翻译: 提供从土壤真菌Coniothyrium minitans获得的新型木聚糖酶基因的核苷酸序列,并表示为cxy1。 还提供了编码的木聚糖酶(称为Cxy1)的氨基酸序列。
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公开(公告)号:US06268202B1
公开(公告)日:2001-07-31
申请号:US09113231
申请日:1998-07-10
IPC分类号: C12N118
CPC分类号: C12Y302/01039 , C12N9/244 , C12R1/645 , C12Y302/01004 , C12Y302/01006 , C12Y302/01058 , C12Y302/01073 , Y10S435/911
摘要: Strains of the aerobic mycoparasitic fungus Coniothyrium minitans having high levels of &bgr;-glucanase activity have been obtained. Preferred strains have been deposited at the American Type Culture Collection under accession numbers 74415, 74416, 74417, 74418, 74419, 74435 and 74436. Methods for mutagenizing C. minitans 74415 and obtaining strains having &bgr;-glucanase activity are provided.
摘要翻译: 已经获得了具有高水平的β-葡聚糖酶活性的需氧真菌寄生真菌菌丝体的菌株。 优选的菌株已经以保藏号74415,74416,74417,74418,74419,74435和74436保藏在美国典型培养物保藏中心。提供了诱变小分子植物74415并获得具有β-葡聚糖酶活性的菌株的方法。
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