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Exploiting Issatchenkia orientalis SD108 for succinic acid production

机译:利用东方伊萨奇尼SD108生产琥珀酸

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Background Recent advances in synthesizing valuable chemicals such as organic acids from low-cost renewable biomass through microbial fermentation have attracted great attention. However, the toxicity of organic acids presents a key challenge to the development of an economically viable fermentation process. Therefore, a platform organism that not only produces organic acids but also tolerates the associated toxicity is highly desirable. Results Here we report the discovery, characterization, and engineering of a yeast strain, Issatchenkia orientalis SD108, that is tolerant to low pH and high concentration of organic acids. This strain demonstrated a higher tolerance compared to I. orientalis ATCC 24210 and Classic Distiller’s Turbo yeast. In order to explore SD108 as a potential platform organism for organic acid production, we determined its draft genome sequence and use the sequencing information to guide pathway design. As proof of concept, an engineered four-gene expression cassette related to the reductive TCA cycle was assembled and integrated into the genome of a uracil auxotroph of SD108. The resulting strain was able to produce succinic acid with a titer of 11.63?g/L, yield of 0.12?g/g, and productivity of 0.11?g/L?·?h in batch cultures using shake flasks. Conclusions The high tolerance of I. orientalis SD108 towards multiple important organic acids makes it a highly attractive organism as a platform host for producing this group of compounds as it will reduce production cost, facilitate downstream processing, and serve as a host for construction of production strains with both pH and specific anion tolerance.
机译:背景技术通过微生物发酵从低成本的可再生生物质中合成有价值的化学物质例如有机酸的最新进展引起了极大的关注。然而,有机酸的毒性对经济上可行的发酵方法的发展提出了关键的挑战。因此,非常需要一种不仅能产生有机酸而且还能耐受相关毒性的平台生物。结果在这里,我们报道了对低pH和高浓度有机酸具有耐受性的酵母Issatchenkia Orientalis SD108的发现,表征和工程改造。与东方I. ATCC 24210和Classic Distiller的Turbo酵母相比,该菌株具有更高的耐受性。为了探索SD108作为生产有机酸的潜在平台生物,我们确定了其基因组序列草案,并使用测序信息来指导途径设计。作为概念的证明,与还原性TCA循环有关的工程化的四基因表达盒被组装并整合到SD108的尿嘧啶营养缺陷型的基因组中。在使用摇瓶的分批培养中,所得菌株能够产生效价为11.63μg/ L,产率为0.12μg/ g,产率为0.11μg/ L··h的琥珀酸。结论东方I. SD108对多种重要有机酸具有高度耐受性,使其成为生产这类化合物的平台宿主,极具吸引力,因为它将降低生产成本,促进下游加工,并作为生产结构的宿主具有pH值和特定阴离子耐受性的菌株。

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