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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
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
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机译：淀粉和木聚糖降解酶表达的淀粉和木聚糖酒精发酵耐热酵母多酚菌株的研制。
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摘要
Genes SWA2 and GAM1 from the yeast, Schwanniomyces occidentalis, encoding -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 -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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机译：分别克隆了来自酵母，西汉产假单胞菌的基因SWA2和GAM1，分别编码-淀粉酶和葡糖淀粉酶。通过将SWA2和GAM1基因整合到多形汉逊酵母染色体中，并与多形汉逊酵母-甘油醛-3-磷酸脱氢酶基因（HpGAP）的强组成型启动子可操作连接，从而实现表达。转化子获得了生长的能力在含有可溶性淀粉作为唯一碳源并且可以在高达10 g / L的高温发酵条件下从乙醇中生产乙醇的基本培养基上。从木霉木霉菌中获得了编码内切木聚糖酶的XYN2基因，而xlnD基因编码为-从木霉黑曲霉中获得木糖苷酶，通过在HpGAP启动子的控制下整合入多形汉逊酵母染色体中来实现这些基因的共表达，所得转化子能够在添加了桦木木聚糖的基本培养基上生长。唯一的碳源。木聚糖水解酶的成功表达导致了能够将桦木木聚糖发酵成乙醇的受体菌株。在48°C的乙醇中。进一步与上述基因在过表达丙酮酸脱羧酶基因的多形汉逊酵母菌株中的共表达进一步改善了乙醇的生产。
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