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    • 59. 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
    • US20110045562A1
    • 2011-02-24
    • US12989735
    • 2009-05-06
    • Charles AbbasAndriy SibirnyAndriy Y. Voronovsky
    • Charles AbbasAndriy SibirnyAndriy Y. Voronovsky
    • C12P7/06C12N1/19C07H21/00C12N15/63
    • C12N9/2428C12N9/2414C12N9/242C12P7/06C12Y302/01001C12Y302/01003C12Y302/01008C12Y302/01037C12Y401/01001Y02E50/17Y02P20/52
    • 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.
    • 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。