Department of Microbiology and Biotechnology, College of Life Sciences, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin, 150030, China.
Department of Strategic and Integrative Research, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
Appl Microbiol Biotechnol. 2024 Nov 5;108(1):503. doi: 10.1007/s00253-024-13342-9.
Enhancing the secretion of recombinant proteins, particularly non-enzymatic proteins that predominate in food and pharmaceutic protein products, remains a significant challenge due to limitations in high-throughput screening methods. This study addresses this bottleneck by establishing a yeast surface display system in the food-grade microorganism Kluyveromyces lactis, enabling efficient display of model target proteins on the yeast cell surface. To assess its potential as a universal high-throughput screening tool for enhanced non-enzymatic protein secretion, we evaluated the consistency between protein display levels and secretion efficiency under the influence of various genetic factors. Our results revealed a strong correlation between these two properties. Furthermore, screening in a random mutagenesis library successfully identified a mutant with improved secretion. These findings demonstrate the potential of the K. lactis surface display system as a powerful and universal tool for high-throughput screening of strains with superior non-enzymatic protein secretion capacity. We believe this study could pave the way for efficient large-scale production of heterologous food and therapeutic proteins in industries. KEY POINTS: • A YSD (yeast surface display) system was established in Kluyveromyces lactis • This system enables high-throughput screening of non-enzymatic protein secretion • This technology assists industrial production of food and therapeutic proteins.
增强重组蛋白的分泌，特别是在食品和药物蛋白产品中占主导地位的非酶蛋白的分泌，仍然是一个重大挑战，因为高通量筛选方法存在局限性。本研究通过在食品级微生物乳酸克鲁维酵母中建立酵母表面展示系统来解决这一瓶颈，使模型靶蛋白能够在酵母细胞表面高效展示。为了评估其作为增强非酶蛋白分泌的通用高通量筛选工具的潜力，我们评估了在各种遗传因素影响下，蛋白展示水平和分泌效率之间的一致性。我们的结果表明这两个特性之间存在很强的相关性。此外，在随机诱变文库中的筛选成功鉴定出一个具有改善分泌能力的突变体。这些发现表明，K. lactis 表面展示系统作为一种强大而通用的高通量筛选工具，具有很高的潜力，可以筛选出具有优异非酶蛋白分泌能力的菌株。我们相信这项研究可以为食品和治疗性蛋白的高效大规模生产铺平道路。 关键点： • 在乳酸克鲁维酵母中建立了 YSD（酵母表面展示）系统 • 该系统能够高通量筛选非酶蛋白的分泌 • 该技术有助于食品和治疗性蛋白的工业生产。
