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Core–Shell Droplet-Based Microfluidic Screening System for Filamentous Fungi
ACS Sensors ( IF 8.2 ) Pub Date : 2023-08-21 , DOI: 10.1021/acssensors.3c01018
Changtai Zhang 1, 2 , Xiaohui Wu 1, 2 , Fuqiang Song 1, 2 , Song Liu 1, 2 , Shiqin Yu 1, 2, 3, 4 , Jingwen Zhou 1, 2, 3, 4
ACS Sensors ( IF 8.2 ) Pub Date : 2023-08-21 , DOI: 10.1021/acssensors.3c01018
Changtai Zhang 1, 2 , Xiaohui Wu 1, 2 , Fuqiang Song 1, 2 , Song Liu 1, 2 , Shiqin Yu 1, 2, 3, 4 , Jingwen Zhou 1, 2, 3, 4
Affiliation
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Filamentous fungi are competitive hosts for the production of drugs, proteins, and chemicals. However, their utility is limited by screening methods and low throughput. In this work, a universal high-throughput system for optimizing protein production in filamentous fungi was described. Droplet microfluidics was used to encapsulate large mutant strain pools in biocompatible core–shell microdroplets designed to avoid mycelial punctures and thus sustain prolonged culture. The self-assembled split GFP was then used to characterize the secretory capacity of the strains and isolate strains with superior production titers according to the fluorescence signals. The platform was applied to optimize the α-amylase secretion of Aspergillus niger, resulting in the isolation of a strain with 2.02-fold higher secretion capacity. The system allows the analysis of >105 single cells per h and will facilitate ultrahigh-throughput screening experiments of filamentous fungi. This method could help identify improved hosts for the large-scale production of biotechnology-relevant proteins. This is a broadly applicable system that can be equally used in other hosts.
中文翻译:
基于核壳液滴的丝状真菌微流控筛选系统
丝状真菌是生产药物、蛋白质和化学品的竞争宿主。然而,它们的实用性受到筛选方法和低通量的限制。在这项工作中,描述了一种用于优化丝状真菌中蛋白质生产的通用高通量系统。液滴微流体用于将大型突变菌株池封装在生物相容性核壳微滴中,旨在避免菌丝穿刺,从而维持长时间培养。然后使用自组装的分裂GFP来表征菌株的分泌能力,并根据荧光信号分离具有优异生产滴度的菌株。该平台用于优化黑曲霉的α-淀粉酶分泌,分离出分泌能力提高2.02倍的菌株。该系统每小时可分析>10 5 个单细胞,并将促进丝状真菌的超高通量筛选实验。该方法可以帮助识别用于大规模生产生物技术相关蛋白质的改良宿主。这是一个适用范围广泛的系统,可以同样用于其他主机。
更新日期:2023-08-21
中文翻译:
![](https://scdn.x-mol.com/jcss/images/paperTranslation.png)
基于核壳液滴的丝状真菌微流控筛选系统
丝状真菌是生产药物、蛋白质和化学品的竞争宿主。然而,它们的实用性受到筛选方法和低通量的限制。在这项工作中,描述了一种用于优化丝状真菌中蛋白质生产的通用高通量系统。液滴微流体用于将大型突变菌株池封装在生物相容性核壳微滴中,旨在避免菌丝穿刺,从而维持长时间培养。然后使用自组装的分裂GFP来表征菌株的分泌能力,并根据荧光信号分离具有优异生产滴度的菌株。该平台用于优化黑曲霉的α-淀粉酶分泌,分离出分泌能力提高2.02倍的菌株。该系统每小时可分析>10 5 个单细胞,并将促进丝状真菌的超高通量筛选实验。该方法可以帮助识别用于大规模生产生物技术相关蛋白质的改良宿主。这是一个适用范围广泛的系统,可以同样用于其他主机。