Centre for Industrial Biotechnology and Biocatalysis (InBio.be), Department of Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000 Ghent, Belgium; Bio Base Europe Pilot Plant (BBEPP), Rodenhuizekaai 1, 9042 Ghent (Desteldonk), Belgium.
Bio Base Europe Pilot Plant (BBEPP), Rodenhuizekaai 1, 9042 Ghent (Desteldonk), Belgium.
Biotechnol Adv. 2023 May-Jun;64:108121. doi: 10.1016/j.biotechadv.2023.108121. Epub 2023 Feb 10.
Recombinant proteins (RP) are widely used as biopharmaceuticals, industrial enzymes, or sustainable food source. Yeasts, with their ability to produce complex proteins through a broad variety of cheap carbon sources, have emerged as promising eukaryotic production hosts. As such, the prevalence of yeasts as favourable production organisms in commercial RP production is expected to increase. Yet, with the selection of a robust production host on the one hand, successful scale-up is dependent on a thorough understanding of the challenging environment and limitations of large-scale bioreactors on the other hand. In the present work, several prominent yeast species, including Saccharomyces cerevisiae, Pichia pastoris, Yarrowia lipolytica, Kluyveromyces lactis and Kluyveromyces marxianus are reviewed for their current state and performance in commercial RP production. Thereafter, the impact of principal process control parameters, including dissolved oxygen, pH, substrate concentration, and temperature, on large-scale RP production are discussed. Finally, technical challenges of process scale-up are identified. To that end, process intensification strategies to enhance industrial feasibility are summarized, specifically highlighting fermentation strategies to ensure sufficient cooling capacity, overcome oxygen limitation, and increase protein quality and productivity. As such, this review aims to contribute to the pursuit of sustainable yeast-based RP production.
重组蛋白（RP）广泛用作生物制药、工业酶或可持续的食物来源。酵母通过广泛的廉价碳源生产复杂蛋白质的能力，已成为很有前途的真核生产宿主。因此，预计酵母作为商业 RP 生产中有利生产生物的流行程度将会增加。然而，一方面需要选择一个强大的生产宿主，另一方面成功地扩大规模取决于对具有挑战性的环境和大型生物反应器的限制有透彻的了解。在本工作中，综述了几种主要的酵母物种，包括酿酒酵母、巴斯德毕赤酵母、解脂耶氏酵母、乳酸克鲁维酵母和马克斯克鲁维酵母，讨论了它们在商业 RP 生产中的现状和性能。然后，讨论了主要过程控制参数（包括溶解氧、pH 值、底物浓度和温度）对大规模 RP 生产的影响。最后，确定了工艺放大的技术挑战。为此，总结了强化工艺的策略以提高工业可行性，特别是强调发酵策略，以确保有足够的冷却能力、克服氧气限制、提高蛋白质质量和生产力。因此，本综述旨在为可持续的基于酵母的 RP 生产提供帮助。
