Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.
Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.
Adv Sci (Weinh). 2023 Oct;10(30):e2302826. doi: 10.1002/advs.202302826. Epub 2023 Aug 30.
Microbial synthesis of valuable hemoproteins has become a popular research topic, and Pichia pastoris is a versatile platform for the industrial production of recombinant proteins. However, the inadequate supply of heme limits the synthesis of high-active hemoproteins. Here a strategy for enhancing intracellular heme biosynthesis to improve the titers and functional activities of hemoproteins is reported. After selecting a suitable expressional strategy for globins, the efficient heme-supply P. pastoris chassis is established by removing the spatial segregation during heme biosynthesis, optimizing precursor synthesis, assembling rate-limiting enzymes using protein scaffolds, and inhibiting heme degradation. This robust chassis produces several highly active hemoproteins, including porcine myoglobin, soy hemoglobin, Vitreoscilla hemoglobin, and P450-BM3, which can be used in the development of artificial meat, high-cell-density fermentation, and whole-cell catalytic synthesis of high-value-added compounds. Furthermore, the engineered chassis strain has great potential for producing and applying other hemoproteins with high activities in various fields.
微生物合成有价值的血红素蛋白已成为一个热门的研究课题，毕赤酵母是工业生产重组蛋白的多功能平台。然而，血红素的供应不足限制了高活性血红素蛋白的合成。本研究报告了一种增强细胞内血红素生物合成的策略，以提高血红素蛋白的效价和功能活性。在选择球蛋白的合适表达策略后，通过去除血红素生物合成过程中的空间隔离、优化前体合成、使用蛋白质支架组装限速酶以及抑制血红素降解，建立了高效血红素供应的毕赤酵母底盘。该底盘能够生产几种高活性的血红素蛋白，包括猪肌红蛋白、大豆血红蛋白、Vitreoscilla 血红蛋白和 P450-BM3，可用于人工肉的开发、高密度发酵和高附加值化合物的全细胞催化合成。此外，该工程底盘菌株在生产和应用其他具有高活性的血红素蛋白方面具有很大的潜力，可应用于各个领域。
