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    • 21. 发明申请
    • GENES CONFERRING TOLERANCE TO ETHANOL AND HIGH TEMPERATURES FOR YEASTS
    • 基于对乙烯和乙烯的高温的承认
    • WO2013106617A2
    • 2013-07-18
    • PCT/US2013021100
    • 2013-01-11
    • ARCHER DANIELS MIDLAND CO
    • ABBAS CHARLESSIBIRNY ANDRIY AVORONOVSKY ANDRIY YISHCHUK OLENA P
    • C12P7/06
    • C12P7/06C07K14/39C07K14/395C12N15/81C12N15/815C12Q1/6811Y02E50/17
    • Methods of identifying genes conferring ethanol tolerance in yeasts, genes that confer ethanol tolerance, and mutant strains used to identify such genes are described. A gene herein designated HpETT1 was isolated from the yeast Hansenula polymorpha. Expression of HpETT1 in an ethanol sensitive mutant H. polymorpha strain designated 7E complimented ethanol sensitivity of the mutant. When multiple copies of the HpETT1 were integrated into the genome and overexpressed, the transformed strain demonstrated approximately 10-fold greater resistance to ethanol and resistance to the protein misfolding agent AZC. Expression of HpETT1 also increased ethanol tolerance in Saccharomyces cerevisiae. HpEtt1 has 39% sequence identity to a previously identified protein from S. cerevisiae denoted MPE1, however, the MPE1 gene does not confer ethanol resistance to the 7E mutant. Another gene from the yeast Pichia stipites was identified that encodes an orthologue protein having 37% identity to HpETT1 herein designated PsETT1 and also confers ethanol resistance to the 7E mutant.
    • 描述赋予酵母中乙醇耐受性的基因,赋予乙醇耐性的基因和用于鉴定这些基因的突变菌株的方法。 本文命名为HpETT1的基因从酵母多形汉逊酵母中分离。 HpETT1在乙醇敏感突变体多形汉逊酵母菌株中的表达命名为7E,突变体的乙醇敏感性。 当HpETT1的多个拷贝整合到基因组中并过表达时,转化的菌株表现出对乙醇的抗性大约为10倍,并且对蛋白质错误折叠剂AZC具有抗性。 HpETT1的表达也增加了酿酒酵母中的乙醇耐受性。 HpEtt1与来自酿酒酵母的先前鉴定的蛋白质具有39%的序列同一性,表示MPE1,然而,MPE1基因不赋予7E突变体的乙醇抗性。 鉴定了来自酵母毕赤酵母属的另一个基因,其编码与本文命名为PsETT1的HpETT1具有37%同一性的直向同源物蛋白,并且还赋予7E突变体的乙醇抗性。
    • 24. 发明申请
    • 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
    • 通过淀粉和XYLAN降解酶的表达可以发现热稳定性YEAST HANSENULA多糖的菌株能够发酵淀粉和XYLAN的醇溶性发酵
    • WO2009137574A3
    • 2010-04-15
    • PCT/US2009042975
    • 2009-05-06
    • ARCHER DANIELS MIDLAND COABBAS CHARLESSIBIRNY ANDRIYVORONOVSKY ANDRIY Y
    • ABBAS CHARLESSIBIRNY ANDRIYVORONOVSKY ANDRIY Y
    • C12N1/19C12N15/56C12P7/06C12R1/01
    • C12N9/2428C12N9/2414C12N9/242C12P7/06C12Y302/01001C12Y302/01003C12Y302/01008C12Y302/01037C12Y401/01001Y02E50/17Y02P20/52
    • Genes SWA2 and GAMl from the yeast, Schwanniomyces occidentalis, encoding a- amylase and glucoamylase, respectively, were cloned and expressed in H. polymorpha. The expression was achieved by integration of the SWA2 and GAMl 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 ß-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.
    • 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。 通过与多形汉逊酵母 - 甘油醛-3-磷酸脱氢酶基因(HpGAP)的强组成型启动子可操作地连接,将SWA2和GAM1基因整合到多形汉逊酵母的染色体中来实现该表达。所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,可以从淀粉高温发酵至10g / L生产乙醇,从真菌木霉菌获得编码内切木聚糖酶的XYN2基因,编码ß的xlnD基因 这些基因的共表达也通过在HpGAP启动子控制下融合到多形汉逊酵母染色体中而得到,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 单独的碳源成功表达木聚糖酶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了乙醇生产。
    • 25. 发明申请
    • CONVERSION OF CARBOHYDRATES TO HYDROXY-METHYLFURFURAL (HMF) AND DERIVATIVES
    • 碳氢化合物转化成羟基甲基纤维素(HMF)和衍生物
    • WO2009076627A3
    • 2009-10-15
    • PCT/US2008086659
    • 2008-12-12
    • ARCHER DANIELS MIDLAND COSANBORN ALEXANDRAHOWARD STEPHEN
    • SANBORN ALEXANDRAHOWARD STEPHEN
    • C07D307/50C07D307/54
    • C07D307/50C07D307/44C07D307/48C07D307/54C07D307/68
    • A method of producing substantially pure HMF, HMF esters and other derivatives from a carbohydrate source by contacting the carbohydrate source with a solid phase catalyst. A carbohydrate starting material is heated in a solvent in a column and continuously flowed through a solid phase catalyst in the presence of an organic acid, or heated with the organic acid and a solid catalyst in solution to form a HMF ester. Heating without organic acid forms HMF. The resulting product is purified by filtration to remove the unreacted starting materials and catalyst. The HMF ester or a mixture of HMF and HMF ester may then be oxidized to 2,5- furandicarboxylic acid (FDCA) by combining the HMF ester with an organic acid, cobalt acetate, manganese acetate and sodium bromide under pressure. Alternatively, the HMF ester may be reduced to form a furan or tetrahydrofuran diol.
    • 通过使碳水化合物源与固相催化剂接触,从碳水化合物源生产基本上纯的HMF,HMF酯和其它衍生物的方法。 将碳水化合物原料在柱中的溶剂中加热,并在有机酸存在下连续流过固相催化剂,或者用有机酸和固体催化剂在溶液中加热形成HMF酯。 无有机酸加热形成HMF。 所得产物通过过滤纯化以除去未反应的起始原料和催化剂。 然后通过在压力下将HMF酯与有机酸,乙酸钴,乙酸锰和溴化钠混合,将HMF酯或HMF和HMF酯的混合物氧化成2,5-呋喃二羧酸(FDCA)。 或者,HMF酯可以还原形成呋喃或四氢呋喃二醇。