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

Paper ID Volume ID Publish Year Pages File Format Full-Text
16832 42616 2016 6 PDF Available
Title
A study on the effects of linker flexibility on acid phosphatase PhoC-GFP fusion protein using a novel linker library
Abstract

•Linker flexibility can efficiently adjust the activities of PhoC-GFP.•Linker with a proper flexibility is necessary to achieve an optimal PhoC-GFP.•Fully rigid or flexible linkers are not the optimal choice for PhoC-GFP.

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 results show that applying a linker library with controllable flexibility to the fusion proteins will be an efficient way to adjust the function of fusion enzymes.

Keywords
Acid phosphatase-green fluorescent protein; Enzyme activity; Fusion protein; Linker flexibility; Linker library
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A study on the effects of linker flexibility on acid phosphatase PhoC-GFP fusion protein using a novel linker library
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 83, February 2016, Pages 1–6
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us