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1. 发明授权

US08323952B2 Alcoholic xylose fermentation at high temperatures by the thermotolerant yeast Hansenula polymorpha 有权
标题翻译：酒精木糖在高温下通过耐热酵母汉氏酵母多形汉逊酵母发酵
公开(公告)号：US08323952B2
公开(公告)日：2012-12-04
申请号：US12330839
申请日：2008-12-09
申请人： Charles A. Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Olena P. Ishchuk
发明人： Charles A. Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Olena P. Ishchuk
IPC分类号： C12N1/00 , C12N1/19
CPC分类号： C12P7/06 , C07K14/39 , C12N9/1205 , C12N9/2402 , C12P7/10 , C12Y207/01017 , C12Y302/01028 , Y02E50/16 , Y02E50/17
摘要： Methods and compositions for the production of ethanol from lignocellulosic starting materials are provided herein. Embodiments provide yeast cells of the genus H. polymorpha with one or more modifications, including, for example, an inactive acid trehalase gene, overexpression of xylulokinase, and/or overexpression of heat-shock protein 104.
摘要翻译：本文提供了用于从木质纤维原料生产乙醇的方法和组合物。实施方案提供多形汉逊酵母属的酵母细胞，其具有一种或多种修饰，包括例如无活性酸海藻糖酶基因，木酮糖激酶的过表达和/或热休克蛋白104的过表达。

2. 发明授权

US09228178B2 Engineering of xylose reductase and overexpression of xylitol dehydrogenase and xylulokinase improves xylose alcoholic fermentation in the thermotolerant yeast Hansenula polymorpha 有权
标题翻译：木糖还原酶的工程和木糖醇脱氢酶和木酮糖激酶的过表达改进了耐热酵母多形汉逊酵母中的木糖酒精发酵
公开(公告)号：US09228178B2
公开(公告)日：2016-01-05
申请号：US12993215
申请日：2009-06-01
申请人： Charles Abbas , Kostyantyn Dmytruk , Olena Dmytruk , Andriy Sibirny , Andriy Y. Voronovsky
发明人： Charles Abbas , Kostyantyn Dmytruk , Olena Dmytruk , Andriy Sibirny , Andriy Y. Voronovsky
IPC分类号： C12P7/06 , C12P7/10 , C12N1/19 , C12N9/12 , C12N9/04
CPC分类号： C12N9/1205 , C12N9/0006 , C12P7/06 , C12P7/10 , C12Y101/01009 , Y02E50/16 , Y02E50/17
摘要： Recombinant genetic constructs and strains of H. polymorpha having significantly increased ethanol productivity with a simultaneous decreased production of xylitol during high-temperature xylose fermentation are disclosed. The constructs include a H. polymorpha XYL1 gene encoding xylose reductase mutated to decrease affinity of the enzyme toward NADPH. The modified version of XYL1 gene under control of a strong constitutive HpGAP promoter was overexpressed in a Δxyl1 background. A recombinant H. polymorpha strain overexpressing the mutated enzyme together with native xylitol dehydrogenase and xylulokinase in the Δxyl1 background was also constructed. Xylose consumption, ethanol and xylitol production by the constructed strain were evaluated during high-temperature xylose fermentation (48° C.). A significant increase in ethanol productivity (up to 7.4 times) was shown in the recombinant strain as compared with the wild type strain. Moreover, the xylitol production by the recombinant strain was reduced considerably: 0.9 mgχ (Lχh)−1 versus 4.2 mgχ (Lxh)−1 by the wild type strain.
摘要翻译：公开了重组遗传构建体和多形汉逊酵母菌株，其在高温木糖发酵过程中同时降低了木糖醇的产生，显着提高了乙醇生产力。构建体包括编码木糖还原酶的多形汉逊酵母XYL1基因突变以降低酶对NADPH的亲和力。在强组成型HpGAP启动子控制下的XYL1基因的修饰版本在一个＆Dgr。xyl1背景中被过表达。还构建了重组多形汉逊酵母菌株，其过表达突变酶与天然木糖醇脱氢酶和木酮糖激酶在＆Dgr; xyl1背景中。在高温木糖发酵（48℃）期间评估了构建菌株的木糖消耗量，乙醇和木糖醇产量。与野生型菌株相比，重组菌株的乙醇生产力显着增加（高达7.4倍）。此外，重组菌株的木糖醇生产显着降低：野生型菌株为0.9mgx（Lχh）-1，而4.2mgx（Lxh）-1）。

3. 发明授权

US08071298B2 Increased ethanol production from xylose 有权
标题翻译：从木糖增加乙醇生产
公开(公告)号：US08071298B2
公开(公告)日：2011-12-06
申请号：US12103861
申请日：2008-04-16
申请人： Charles Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Oleh V. Stasyk , Olena P. Ishchuk , Olena B. Ryabova
发明人： Charles Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Oleh V. Stasyk , Olena P. Ishchuk , Olena B. Ryabova
IPC分类号： C12Q1/68 , C12N15/11 , C12N1/19
CPC分类号： C12P7/10 , C12N9/88 , Y02E50/16 , Y02E50/17
摘要： Methods and compositions for the production of ethanol from lignocellulosic starting materials are provided herein. Embodiments of the invention provide methods of manipulating the carbon flux of a host cell transformed with plasmids of the invention. Plasmids of the invention may include nucleotides that encode pyruvate decarboxylase. In one embodiment, a strain of the thermotolerant yeast Hansenula polymorpha that has been transformed with plasmids and polynucleotides of the invention is provided.
摘要翻译：本文提供了用于从木质纤维原料生产乙醇的方法和组合物。本发明的实施方案提供了操纵用本发明的质粒转化的宿主细胞的碳通量的方法。本发明的质粒可以包括编码丙酮酸脱羧酶的核苷酸。在一个实施方案中，提供已经用本发明的质粒和多核苷酸转化的耐热酵母汉逊酵母多形汉逊菌株。

4. 发明授权

US09388399B2 Development of strains of the thermotolerant yeast Hansenula polymorpha capable of alcoholic fermentation of starch and xylan by expression of starch and xylan degrading enzymes 有权
标题翻译：通过淀粉和木聚糖降解酶的表达开发能够通过淀粉和木聚糖进行酒精发酵的耐热酵母汉逊氏酵母菌株
公开(公告)号：US09388399B2
公开(公告)日：2016-07-12
申请号：US12989735
申请日：2009-05-06
申请人： Charles Abbas , Andriy Sibirny , Andriy Y. Voronovsky
发明人： Charles Abbas , Andriy Sibirny , Andriy Y. Voronovsky
IPC分类号： C12N9/34 , C12N9/28 , C12P7/06 , C12N9/30 , C12N9/26
CPC分类号： C12N9/2428 , C12N9/2414 , C12N9/242 , C12P7/06 , C12Y302/01001 , C12Y302/01003 , C12Y302/01008 , C12Y302/01037 , C12Y401/01001 , Y02E50/17 , Y02P20/52
摘要： Genes SWA2 and GAM1 from the yeast, Schwanniomyces occidentalis, encoding α-amylase and glucoamylase, respectively, were cloned and expressed in H. polymorpha. The expression was achieved by integration of the SWA2 and GAM1 genes into the chromosome of H. polymorpha under operably linked to a strong constitutive promoter of the H. polymorpha-glyceraldehyde-3-phosphate dehydrogenase gene (HpGAP. Resulting transformants acquired the ability to grow on a minimal medium containing soluble starch as a sole carbon source and can produce Ethanol at high-temperature fermentation from starch up to 10 g/L. A XYN2 gene encoding endoxylanase was obtained from the fungus Trichoderma resee, and a xlnD gene coding for β-xylosidase was obtained from the fungus Aspergillus niger. Co-expression of these genes was also achieved by integration into the H. polymorpha chromosome under control of the HpGAP promoter. The resulting transformants were capable of growth on a minimal medium supplemented with birchwood xylan as a sole carbon source. Successful expression of xylanolytic enzymes resulted in a recipient strain capable of fermentation of birchwood xylan to ethanol at 48° C. Further with co expression of the forgoing genes in a H. polymorpha strain that overexpresses a pyruvate decarboxylase gene further improved ethanol production.
摘要翻译：来自酵母的SWA2和GAM1基因分别编码α-淀粉酶和葡糖淀粉酶，分别在多形汉逊酵母中表达。该表达是通过将SWA2和GAM1基因整合到多形汉逊酵母的染色体中而实现的，其可操作地连接于多形汉逊酵母 - 甘油醛-3-磷酸脱氢酶基因（HpGAP）的强组成型启动子。得到的转化体获得了生长能力在含有可溶性淀粉作为唯一碳源的基本培养基上，可以从高达10g / L的淀粉高温发酵生产乙醇，从真菌木霉菌获得编码内切木聚糖酶的XYN2基因，并获得编码＆bgr的xlnD基因来自真菌黑曲霉的α-脱氧核苷酸酶，这些基因的共表达也通过在HpGAP启动子控制下整合入多形汉逊酵母染色体而实现，所得转化体能够在补充有桦木木聚糖的基本培养基上生长作为唯一的碳源，木聚糖酶的成功表达导致能够发酵桦木木聚糖的受体菌株到48℃的乙醇。此外，在多克隆多因子菌株中共表达过去基因，其过表达丙酮酸脱羧酶基因进一步改善乙醇生产。

5. 发明申请

US20110045562A1 Development of Strains of the Thermotolerant Yeast Hansenula Polymorpha Capable of Alcoholic Fermentation of Starch and Xylan by Expression of Starch and Xylan Degrading Enzymes 有权
标题翻译：通过淀粉和木聚糖降解酶的表达开发可以通过淀粉和木聚糖的酒精发酵的耐热酵母汉逊酵母多形酵菌菌株
公开(公告)号：US20110045562A1
公开(公告)日：2011-02-24
申请号：US12989735
申请日：2009-05-06
申请人： Charles Abbas , Andriy Sibirny , Andriy Y. Voronovsky
发明人： Charles Abbas , Andriy Sibirny , Andriy Y. Voronovsky
IPC分类号： C12P7/06 , C12N1/19 , C07H21/00 , C12N15/63
CPC分类号： C12N9/2428 , C12N9/2414 , C12N9/242 , C12P7/06 , C12Y302/01001 , C12Y302/01003 , C12Y302/01008 , C12Y302/01037 , C12Y401/01001 , Y02E50/17 , Y02P20/52
摘要： Genes SWA2 and GAMl from the yeast, Schwanniomyces occidentalis, encoding α-amylase and glucoamylase, respectively, were cloned and expressed in H. polymorpha. The expression was achieved by integration of the SWA2 and GAM1 genes into the chromosome of H. polymorpha under operably linked to a strong constitutive promoter of the H. polymorpha-glyceraldehyde-3-phosphate dehydrogenase gene (HpGAP. Resulting transformants acquired the ability to grow on a minimal medium containing soluble starch as a sole carbon source and can produce Ethanol at high-temperature fermentation from starch up to 10 g/L. A XYN2 gene encoding endoxylanase was obtained from the fungus Trichoderma resee, and a xlnD gene coding for β-xylosidase was obtained from the fungus Aspergillus niger. Co-expression of these genes was also achieved by integration into the H. polymorpha chromosome under control of the HpGAP promoter. The resulting transformants were capable of growth on a minimal medium supplemented with birchwood xylan as a sole carbon source. Successful expression of xylanolytic enzymes resulted in a recipient strain capable of fermentation of birchwood xylan to ethanol at 48° C. Further with co expression of the for-going genes in a H. polymorpha strain that overexpresses a pyruvate decarboxylase gene further improved ethanol production.
摘要翻译：来自酵母的基因SWA2和GAM1分别编码α-淀粉酶和葡糖淀粉酶，分别在多形汉逊酵母中表达。该表达是通过将SWA2和GAM1基因整合到多形汉逊酵母的染色体中而实现的，其可操作地连接于多形汉逊酵母 - 甘油醛-3-磷酸脱氢酶基因（HpGAP）的强组成型启动子。得到的转化体获得了生长能力在含有可溶性淀粉作为唯一碳源的基本培养基上，可以从高达10g / L的淀粉高温发酵生产乙醇，从真菌木霉菌获得编码内切木聚糖酶的XYN2基因，并获得编码＆bgr的xlnD基因来自真菌黑曲霉的α-脱氧核苷酸酶，这些基因的共表达也通过在HpGAP启动子控制下整合入多形汉逊酵母染色体而实现，所得转化体能够在补充有桦木木聚糖的基本培养基上生长作为唯一的碳源，木聚糖酶的成功表达导致能够发酵桦木木聚糖的受体菌株到48℃的乙醇。此外，过表达丙酮酸脱羧酶基因的多形汉逊酵母菌株中正在进行的基因的共表达进一步改进了乙醇生产。

6. 发明申请

US20110086402A1 ENGINEERING OF XYLOSE REDUCTASE AND OVEREXPRESSION OF XYLITOL DEHYDROGENASE AND XYLULOKINASE IMPROVES XYLOSE ALCOHOLIC FERMENTATION IN THE THERMOTOLERANT YEAST HANSENULA POLYMORPHA 审中-公开
标题翻译： XYLOSE还原酶的工程和XYLITOL DEHYDROGENASE和XYLULOKINASE的过表达改善了热稳定剂YANSTER HANSENULA POLYMORPHA中的XYLOSE酒精发酵
公开(公告)号：US20110086402A1
公开(公告)日：2011-04-14
申请号：US12993215
申请日：2009-06-01
申请人： Charles Abbas , Kostyantyn Dmytruk , Olena Dmytruk , Andriy Sibirny , Andriy Y. Voronovsky
发明人： Charles Abbas , Kostyantyn Dmytruk , Olena Dmytruk , Andriy Sibirny , Andriy Y. Voronovsky
IPC分类号： C12P7/06 , C12N1/19 , C07H21/00 , C07H21/04
CPC分类号： C12N9/1205 , C12N9/0006 , C12P7/06 , C12P7/10 , C12Y101/01009 , Y02E50/16 , Y02E50/17
摘要： Recombinant genetic constructs and strains of H. polymorpha having significantly increased ethanol productivity with a simultaneous decreased production of xylitol during high-temperature xylose fermentation are disclosed. The constructs include a H. polymorpha XYL1 gene encoding xylose reductase mutated to decrease affinity of the enzyme toward NADPH. The modified version of XYL1 gene under control of a strong constitutive HpGAP promoter was overexpressed in a Δxyll background. A recombinant H. polymorpha strain overexpressing the mutated enzyme together with native xylitol dehydrogenase and xylulokinase in the Δxyll background was also constructed. Xylose consumption, ethanol and xylitol production by the constructed strain were evaluated during high-temperature xylose fermentation (48° C.). A significant increase in ethanol productivity (up to 7.4 times) was shown in the recombinant strain as compared with the wild type strain. Moreover, the xylitol production by the recombinant strain was reduced considerably: 0.9 mgχ (Lχh)−1 versus 4.2 mgχ (Lxh)−1 by the wild type strain.
摘要翻译：公开了重组遗传构建体和多形汉逊酵母菌株，其在高温木糖发酵过程中同时降低了木糖醇的产生，显着提高了乙醇生产力。构建体包括编码木糖还原酶的多形汉逊酵母XYL1基因突变以降低酶对NADPH的亲和力。在强组成型HpGAP启动子的控制下，修饰版本的XYL1基因在一个＆Dgr; xyll背景中被过表达。还构建了在多克隆背景中过表达突变酶与天然木糖醇脱氢酶和木酮糖激酶的重组多形汉逊酵母菌株。在高温木糖发酵（48℃）期间评估了构建菌株的木糖消耗量，乙醇和木糖醇产量。与野生型菌株相比，重组菌株的乙醇生产力显着增加（高达7.4倍）。此外，重组菌株的木糖醇生产显着降低：野生型菌株为0.9mgx（Lχh）-1，而4.2mgx（Lxh）-1）。

7. 发明申请

US20090155872A1 ALCOHOLIC XYLOSE FERMENTATION AT HIGH TEMPERATURES BY THE THERMOTOLERANT YEAST HANSENULA POLYMORPHA 有权
标题翻译：热稳定剂YYSAST HANSENULA POLYMORPHA在高温下的酒精性半乳糖发酵
公开(公告)号：US20090155872A1
公开(公告)日：2009-06-18
申请号：US12330839
申请日：2008-12-09
申请人： Charles A. Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Olena P. Ishchuk
发明人： Charles A. Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Olena P. Ishchuk
IPC分类号： C12P7/06 , C12N1/19
CPC分类号： C12P7/06 , C07K14/39 , C12N9/1205 , C12N9/2402 , C12P7/10 , C12Y207/01017 , C12Y302/01028 , Y02E50/16 , Y02E50/17
摘要： Methods and compositions for the production of ethanol from lignocellulosic starting materials are provided herein. Embodiments provide yeast cells of the genus H. polymorpha with one or more modifications, including, for example, an inactive acid trehalase gene, overexpression of xylulokinase, and/or overexpression of heat-shock protein 104.
摘要翻译：本文提供了用于从木质纤维原料生产乙醇的方法和组合物。实施方案提供多形汉逊酵母属的酵母细胞，其具有一种或多种修饰，包括例如无活性酸海藻糖酶基因，木酮糖激酶的过表达和/或热休克蛋白104的过表达。

8. 发明申请

US20080254524A1 Increased Ethanol Production from Xylose 有权
标题翻译：从木糖增加乙醇生产
公开(公告)号：US20080254524A1
公开(公告)日：2008-10-16
申请号：US12103861
申请日：2008-04-16
申请人： Charles Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Oleh V. Stasyk , Olena P. Ishchuk , Olena B. Ryabova
发明人： Charles Abbas , Andriy A. Sibirny , Andriy Y. Voronovsky , Oleh V. Stasyk , Olena P. Ishchuk , Olena B. Ryabova
IPC分类号： C12P7/06 , C07H21/04 , C12N15/00 , C12N1/00
CPC分类号： C12P7/10 , C12N9/88 , Y02E50/16 , Y02E50/17
摘要： Methods and compositions for the production of ethanol from lignocellulosic starting materials are provided herein. Embodiments of the invention provide methods of manipulating the carbon flux of a host cell transformed with plasmids of the invention. Plasmids of the invention may include nucleotides that encode pyruvate decarboxylase. In one embodiment, a strain of the thermotolerant yeast Hansenula polymorpha that has been transformed with plasmids and polynucleotides of the invention is provided.
摘要翻译：本文提供了用于从木质纤维原料生产乙醇的方法和组合物。本发明的实施方案提供了操纵用本发明的质粒转化的宿主细胞的碳通量的方法。本发明的质粒可以包括编码丙酮酸脱羧酶的核苷酸。在一个实施方案中，提供已经用本发明的质粒和多核苷酸转化的耐热酵母汉逊酵母多形汉逊菌株。

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