Invention Grant
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
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
- Patent Title: 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
- Patent Title (中): 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
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Application No.: US12989735Application Date: 2009-05-06
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Publication No.: US09388399B2Publication Date: 2016-07-12
- Inventor: Charles Abbas , Andriy Sibirny , Andriy Y. Voronovsky
- Applicant: Charles Abbas , Andriy Sibirny , Andriy Y. Voronovsky
- Applicant Address: US IL Decatur
- Assignee: Archer Daniels Midland Company
- Current Assignee: Archer Daniels Midland Company
- Current Assignee Address: US IL Decatur
- Agent Corey Crafton
- International Application: PCT/US2009/042975 WO 20090506
- International Announcement: WO2009/137574 WO 20091112
- Main IPC: C12N9/34
- IPC: C12N9/34 ; C12N9/28 ; C12P7/06 ; C12N9/30 ; C12N9/26

Abstract:
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.
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