Mariah B M J Kes, Biwen Wang, Peter van Ulsen, Leendert W Hamoen, Joen Luirink
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引用次数: 0
摘要
枯草杆菌（Bacillus subtilis）是一种革兰氏阳性细菌，因其卓越的蛋白质分泌能力，经常被生物工业用于生产各种蛋白质。要确定枯草杆菌分泌蛋白质的最佳条件，快速灵敏的蛋白质分泌测量方法至关重要。快速、通用的检测方法对于高通量检测各种蛋白质最为有用。在本研究中，我们介绍了一种基于分离荧光素酶的方法来测量枯草杆菌的蛋白质分泌。NanoBiT 系统用于监测四种不同蛋白质的分泌：木聚糖酶 A、淀粉酶 M、谷氨酰胺酶 A 蛋白和 GFP 纳米抗体。我们的研究结果表明，分裂荧光素酶系统是一种快速、灵敏、用户友好的方法。
Development of a split-luciferase assay to establish optimal protein secretion conditions for protein production by Bacillus subtilis.
Bacillus subtilis is a Gram-positive bacterium that is frequently used in the bioindustry for the production of various proteins, because of its superior protein secretion capacities. To determine optimal conditions for protein secretion by B. subtilis, a quick and sensitive method for measuring protein secretion is crucial. A fast and universal assay is most useful for detecting diverse proteins in a high-throughput manner. In this study, we introduce a split-luciferase-based method for measuring protein secretion by B. subtilis. The NanoBiT system was used to monitor secretion of four different proteins: xylanase A, amylase M, protein glutaminase A, and GFP nanobody. Our findings underscore the split-luciferase system as a quick, sensitive, and user-friendly method.
期刊介绍：
We publish high-quality original research on bacteria, fungi, protists, archaea, algae, parasites and other microscopic life forms.
Topics include but are not limited to:
Antimicrobials and antimicrobial resistance
Bacteriology and parasitology
Biochemistry and biophysics
Biofilms and biological systems
Biotechnology and bioremediation
Cell biology and signalling
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Cross-disciplinary work
Ecology and environmental microbiology
Food microbiology
Genetics
Host–microbe interactions
Microbial methods and techniques
Microscopy and imaging
Omics, including genomics, proteomics and metabolomics
Physiology and metabolism
Systems biology and synthetic biology
The microbiome.
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