TY - JOUR
T1 - Discovery of Novel Pseudomonas putida Flavin-Binding Fluorescent Protein Variants with Significantly Improved Quantum Yield
AU - Ko, Sanghwan
AU - Ko, Sanghwan
AU - Jeon, Hyunwoo
AU - Yoon, Sanghan
AU - Kyung, Munsu
AU - Yun, Hyungdon
AU - Na, Jung Hyun
AU - Jung, Sang Taek
N1 - Funding Information:
This work was supported by a Korea University grant and grants from the Basic Science Research Program (2019R1F1A1059834 and 2019R1A4A1029000), the Bio and Medical Technology Development Program (2017M3A9C8060541), and the Pioneer Research Center Program (2014M3C1A3051460) through the National Research Foundation of Korea funded by the Ministry of Science and ICT.
Publisher Copyright:
© 2020 American Chemical Society.
PY - 2020/5/27
Y1 - 2020/5/27
N2 - 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.
AB - 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.
KW - flavin-binding fluorescent protein
KW - fluorescence intensity
KW - quantum yield
UR - http://www.scopus.com/inward/record.url?scp=85085420049&partnerID=8YFLogxK
U2 - 10.1021/acs.jafc.0c00121
DO - 10.1021/acs.jafc.0c00121
M3 - Article
C2 - 32367716
AN - SCOPUS:85085420049
VL - 68
SP - 5873
EP - 5879
JO - Journal of Agricultural and Food Chemistry
JF - Journal of Agricultural and Food Chemistry
SN - 0021-8561
IS - 21
ER -