Higher alcohol isobutanol is a promising liquid fuel. During alcoholic fermentation, Saccharomyces cerevisiae produces only trace amounts of isobutanol. Screening the collection of nonconventional yeasts show that Magnusiomyces magnusii accumulates 440 mg of isobutanol per L in rich YPD medium. Here, the transformation protocol for M. magnusii is adapted based on the use of the dominant markers conferring resistance to nourseothricin or zeocin; the strong constitutive promoter TEF1 is cloned and a reporter system based on LAC4 gene from Kluyveromyces lactis coding for β‐galactosidase is constructed. In order to increase isobutanol production in M. magnusii , the heterologous gene ILV2 from S. cerevisiae is expressed in M. magnusii under control of the TEF1 promoter. The best stabilized transformants produce 620 mg of isobutanol per L in YPD medium and 760 mg L⁻¹ in the medium with 2‐oxoisovalerate. This suggests that M. magnusii is a promising organism for further development of a robust isobutanol producer.

中文翻译：

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多核酵母菌属（Magnusii）（Dipodascus，Endomyces）是一种有希望的异丁醇生产者。

高级醇异丁醇是有前途的液体燃料。在酒精发酵过程中，酿酒酵母仅产生痕量的异丁醇。筛选非常规酵母的集合表明，Magnusiomyces magnusii在富YPD培养基中每L累积440 mg异丁醇。这里，对于转化方案M. magnusii适于基于使用的显性标记赋予对诺尔丝菌素或Zeocin抗性的; 克隆了强组成型启动子TEF1，并构建了一个基于乳酸克鲁维酵母LAC4基因的报告系统，该系统编码β-半乳糖苷酶。为了增加马格纳霉中的异丁醇产量中，异源基因ILV2从酿酒酵母中表达的M. magnusii下的控制TEF1启动子。最佳稳定的转化子在YPD培养基中每L产生620 mg异丁醇，在含有2-氧代异戊酸的培养基中产生760 mg L ^-1。这表明马格纳霉菌是有希望的生物体，可用于进一步发展健壮的异丁醇生产商。