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Coproduction of Phase-Separated Carotenoids and β-Farnesene as a Yeast Biomass Valorization Strategy
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2023-09-18 , DOI: 10.1021/acssynbio.3c00270 Yoseph Tsegaye 1 , Phoebe Yeh 1 , Victor Holmes 1 , Matthew Jones 1 , Alexander Kilbo 1 , Chris N Micklem 1 , Chia-Hong Tsai 1 , Christopher J Paddon 1
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2023-09-18 , DOI: 10.1021/acssynbio.3c00270 Yoseph Tsegaye 1 , Phoebe Yeh 1 , Victor Holmes 1 , Matthew Jones 1 , Alexander Kilbo 1 , Chris N Micklem 1 , Chia-Hong Tsai 1 , Christopher J Paddon 1
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Valorization, the process whereby waste materials are converted into more valuable products, is rarely practiced in industrial fermentation. We developed a model valorization system whereby Saccharomyces cerevisiae that had previously been engineered to produce high concentrations (>100 g/L) of extracellular β-farnesene was further engineered to simultaneously produce intracellular carotenoids, both products being isoprenoids. Thus, a single fermentation generates two valuable products, namely, β-farnesene in the liquid phase and carotenoids in the solid biomass phase. Initial attempts to produce high levels of canthaxanthin (a ketocarotenoid used extensively in animal feed) in a β-farnesene production strain negatively impacted both biomass growth and β-farnesene production. A refined approach used a promoter titration strategy to reduce β-carotene production to a level that had minimal impact on growth and β-farnesene production in fed-batch fermentations and then engineered the resulting strain to produce canthaxanthin. Further optimization of canthaxanthin coproduction used a bioprospecting approach to identify ketolase enzymes that maximized conversion of β-carotene to canthaxanthin. Finally, we demonstrated that β-carotene is not present in the extracellular β-farnesene at a significant concentration and that which is present can be removed by a simple distillation, indicating that β-farnesene (the primary fermentation product) purity is unaffected by coproduction of carotenoids.
中文翻译：
相分离类胡萝卜素和 β-金合欢烯的联产作为酵母生物质增值策略
增值是将废物转化为更有价值的产品的过程，在工业发酵中很少进行。我们开发了一个模型增值系统，通过该系统，先前被改造为生产高浓度（>100 g/L）细胞外β-法呢烯的酿酒酵母被进一步改造为同时生产细胞内类胡萝卜素，这两种产品都是类异戊二烯。因此，一次发酵产生两种有价值的产物，即液相中的β-法呢烯和固体生物质相中的类胡萝卜素。最初尝试在 β-法呢烯生产菌株中生产高水平的角黄素（一种广泛用于动物饲料的酮类胡萝卜素），但对生物量增长和 β-法呢烯生产产生了负面影响。一种改进的方法使用启动子滴定策略将 β-胡萝卜素产量降低到对补料分批发酵中的生长和 β-法呢烯产量影响最小的水平，然后对所得菌株进行改造以生产角黄素。角黄素共生产的进一步优化使用生物勘探方法来鉴定能够最大限度地将 β-胡萝卜素转化为角黄素的酮酶。最后，我们证明胞外β-法呢烯中不存在高浓度的β-胡萝卜素，并且可以通过简单的蒸馏除去存在的β-胡萝卜素，这表明β-法呢烯（初级发酵产物）的纯度不受联产的影响类胡萝卜素。
更新日期：2023-09-18
中文翻译：
相分离类胡萝卜素和 β-金合欢烯的联产作为酵母生物质增值策略
增值是将废物转化为更有价值的产品的过程，在工业发酵中很少进行。我们开发了一个模型增值系统，通过该系统，先前被改造为生产高浓度（>100 g/L）细胞外β-法呢烯的酿酒酵母被进一步改造为同时生产细胞内类胡萝卜素，这两种产品都是类异戊二烯。因此，一次发酵产生两种有价值的产物，即液相中的β-法呢烯和固体生物质相中的类胡萝卜素。最初尝试在 β-法呢烯生产菌株中生产高水平的角黄素（一种广泛用于动物饲料的酮类胡萝卜素），但对生物量增长和 β-法呢烯生产产生了负面影响。一种改进的方法使用启动子滴定策略将 β-胡萝卜素产量降低到对补料分批发酵中的生长和 β-法呢烯产量影响最小的水平，然后对所得菌株进行改造以生产角黄素。角黄素共生产的进一步优化使用生物勘探方法来鉴定能够最大限度地将 β-胡萝卜素转化为角黄素的酮酶。最后，我们证明胞外β-法呢烯中不存在高浓度的β-胡萝卜素，并且可以通过简单的蒸馏除去存在的β-胡萝卜素，这表明β-法呢烯（初级发酵产物）的纯度不受联产的影响类胡萝卜素。
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