TY - JOUR
T1 - Engineered Arabidopsis Blue Light Receptor LOV Domain Variants with Improved Quantum Yield, Brightness, and Thermostability
AU - Ko, Sanghwan
AU - Hwang, Bora
AU - Na, Jung Hyun
AU - Lee, Jisun
AU - Jung, Sang Taek
N1 - Funding Information:
This work was supported by grants from the Basic Science Research Program (2019R1F1A1059834, 2019R1A4A1029000, and 2018R1A6A3A11050730), the Bio & 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:
© 2019 American Chemical Society.
PY - 2019
Y1 - 2019
N2 - Despite remarkable contribution of green fluorescent protein and its variants for better understanding of various biological functions, its application for anaerobic microorganisms has been limited because molecular oxygen is essential for chromophore formation. To overcome the limitation, we engineered a plant-derived light, oxygen, or voltage (LOV) domain containing flavin mononucleotide for enhanced spectral properties. The resulting LOV variants exhibited improved fluorescence intensity (20 and 70% higher for SH3 and 70% for BR1, respectively) compared to iLOV, an LOV variant isolated in a previous study, and the quantum yields of the LOV variants (0.40 for SH3 and 0.45 for BR1) were also improved relative to that of iLOV (Q = 0.37). In addition to fluorescence intensity, the identified mutations of SH3 enabled an improved thermostability of the protein. The engineered LOV variants with enhanced spectral properties could provide a valuable tool for fluorescent molecular probes under anaerobic conditions.
AB - Despite remarkable contribution of green fluorescent protein and its variants for better understanding of various biological functions, its application for anaerobic microorganisms has been limited because molecular oxygen is essential for chromophore formation. To overcome the limitation, we engineered a plant-derived light, oxygen, or voltage (LOV) domain containing flavin mononucleotide for enhanced spectral properties. The resulting LOV variants exhibited improved fluorescence intensity (20 and 70% higher for SH3 and 70% for BR1, respectively) compared to iLOV, an LOV variant isolated in a previous study, and the quantum yields of the LOV variants (0.40 for SH3 and 0.45 for BR1) were also improved relative to that of iLOV (Q = 0.37). In addition to fluorescence intensity, the identified mutations of SH3 enabled an improved thermostability of the protein. The engineered LOV variants with enhanced spectral properties could provide a valuable tool for fluorescent molecular probes under anaerobic conditions.
KW - directed evolution
KW - flavin-binding fluorescent protein
KW - fluorescence intensity
KW - quantum yield
UR - http://www.scopus.com/inward/record.url?scp=85073822274&partnerID=8YFLogxK
U2 - 10.1021/acs.jafc.9b05473
DO - 10.1021/acs.jafc.9b05473
M3 - Article
C2 - 31581772
AN - SCOPUS:85073822274
SN - 0021-8561
JO - Journal of agricultural and food chemistry
JF - Journal of agricultural and food chemistry
ER -