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Engineering the thermotolerant industrial yeast Kluyveromyces marxianus for anaerobic growth
Metabolic Engineering ( IF 8.4 ) Pub Date : 2021-07-23 , DOI: 10.1016/j.ymben.2021.07.006
Wijbrand J C Dekker 1 , Raúl A Ortiz-Merino 1 , Astrid Kaljouw 1 , Julius Battjes 1 , Frank W Wiering 1 , Christiaan Mooiman 1 , Pilar de la Torre 1 , Jack T Pronk 1
Affiliation  

Current large-scale, anaerobic industrial processes for ethanol production from renewable carbohydrates predominantly rely on the mesophilic yeast Saccharomyces cerevisiae. Use of thermotolerant, facultatively fermentative yeasts such as Kluyveromyces marxianus could confer significant economic benefits. However, in contrast to S. cerevisiae, these yeasts cannot grow in the absence of oxygen. Responses of K. marxianus and S. cerevisiae to different oxygen-limitation regimes were analyzed in chemostats. Genome and transcriptome analysis, physiological responses to sterol supplementation and sterol-uptake measurements identified absence of a functional sterol-uptake mechanism as a key factor underlying the oxygen requirement of K. marxianus. Heterologous expression of a squalene-tetrahymanol cyclase enabled oxygen-independent synthesis of the sterol surrogate tetrahymanol in K. marxianus. After a brief adaptation under oxygen-limited conditions, tetrahymanol-expressing K. marxianus strains grew anaerobically on glucose at temperatures of up to 45 °C. These results open up new directions in the development of thermotolerant yeast strains for anaerobic industrial applications.



中文翻译:

工程耐热工业酵母克鲁维酵母马克斯酵母厌氧生长

目前从可再生碳水化合物生产乙醇的大规模厌氧工业过程主要依赖于嗜温酵母酿酒酵母。使用耐热的兼性发酵酵母如马克斯克鲁维酵母可以带来显着的经济效益。然而,与酿酒酵母不同的是,这些酵母菌在缺氧的情况下无法生长。响应马克斯克鲁维酵母酿酒酵母在恒化器中分析了对不同限氧机制的影响。基因组和转录组分析、对甾醇补充的生理反应和甾醇摄取测量表明,缺乏功能性甾醇摄取机制是马克斯 K. marxianus需氧量的关键因素。角鲨烯-四氢甘醇环化酶的异源表达使马克斯金枪鱼中甾醇替代物四氢甘醇的非氧依赖性合成成为可能。在氧气有限的条件下短暂适应后,表达四氢甘醇的马氏 K. marxianus菌株在高达 45 °C 的温度下在葡萄糖上厌氧生长。这些结果为开发用于厌氧工业应用的耐热酵母菌株开辟了新的方向。

更新日期:2021-08-01
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