Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 68, Issue 21, Pages 5873-5879Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.0c00121
Keywords
flavin-binding fluorescent protein; fluorescence intensity; quantum yield
Funding
- Korea University grant
- Basic Science Research Program through the National Research Foundation of Korea - Ministry of Science and ICT [2019R1F1A1059834, 2019R1A4A1029000]
- Bio and Medical Technology Develoment Program through the National Research Foundation of Korea - Ministry of Science and ICT [2017M3A9C8060541]
- Pioneer Research Center Program through the National Research Foundation of Korea - Ministry of Science and ICT [2014M3C1A3051460]
- National Research Foundation of Korea [2019R1F1A1059834] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Oxygen-independent, flavin-binding fluorescent proteins (FbFPs) are emerging as alternatives to green fluorescent protein (GFP), which has limited applicability in studying anaerobic microorganisms, such as human gastrointestinal bacteria, which grow in oxygen-deficient environments. However, the utility of these FbFPs has been compromised because of their poor fluorescence emission. To overcome this limitation, we have employed a high-throughput library screening strategy and engineered an FbFP derived from Pseudomonas putida (SB2) for enhanced quantum yield. Of the resulting SB2 variants, KOFP-7 exhibited a significantly improved quantum yield (0.61) compared to other reported engineered FbFPs, which was even higher than that of enhanced GFP (EGFP, 0.60), with significantly enhanced tolerance against a strong reducing agent.
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