A study on the effects of linker flexibility on acid phosphatase PhoC-GFP fusion protein using a novel linker library

Enzyme and Microbial Technology
Ziliang HuangXin-Hui Xing

Abstract

Fusion strategy has been widely used to construct artificial multifunction proteins. The flexibility or rigidity of linkers between two fused partners is an important parameter that affects the function of fusion proteins. By combining the flexible unit GGGGS (F) and rigid unit EAAAK (R), ten linkers consisting of five elementary units that cover the fully rigid RRRRR linker to the fully flexible FFFFF linker were used to construct acid phosphatase-green fluorescence protein fusion protein (PhoC-GFP). By varying the linker flexibility in PhoC-GFPs, the relative specific activity of phosphotransferase and phosphatase varied from ∼19.0% to 100% and ∼9.35% to 100%, respectively. There exists an optimal linker capable of achieving the highest phosphotransferase/phosphatase activity and GFP fluorescence intensity. We found that the highest activities were achieved neither with the rigid RRRRR linker nor with the flexible FFFFF linker, but with the FFFRR linker. Linker flexibility could adjust the activity ratio between phosphotransferase and phosphatase and varied between ∼30% to 100%. PhoC-GFP with FRRRR linker achieved the highest relative specific phosphotransferase activity/relative specific phosphatase activity (T/P) value. Our...Continue Reading

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Citations

Aug 3, 2020·Biotechnology and Bioengineering·Xiaoqiang JiaKang Wu
Nov 16, 2016·Frontiers in Cellular and Infection Microbiology·Víctor M Luna-PinedaJuan Xicohtencatl-Cortes
Jan 29, 2020·Applied Microbiology and Biotechnology·Ping LinTengfei Wang
Mar 24, 2021·International Journal of Biological Macromolecules·Junling GuoZhemin Zhou
Aug 12, 2021·Scientific Reports·Jongjin ParkSunghwa Choe

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