Effect of flexible linker length on the activity of fusion protein 4-coumaroyl-CoA ligase::stilbene synthase.

Abstract:

:In order to elucidate the effect of flexible linker length on the catalytic efficiency of fusion proteins, two short flexible peptide linkers of various lengths were fused between Arabidopsis thaliana 4-coumaroyl-CoA ligase (4CL) and Polygonum cuspidatum stilbene synthase (STS) to generate fusion proteins 4CL-(GSG)n-STS (n ≤ 5) and 4CL-(GGGGS)n-STS (n ≤ 4). The fusion proteins were expressed in both Escherichia coli and Saccharomyces cerevisiae, and their bioactivities were tested in vitro and in vivo using purified proteins and engineered strains, respectively. The catalytic efficiency of the fusions decreased gradually with the increase of GSG or GGGGS repeats. In both engineered S. cerevisiae and E. coli in vivo experiments, the capacity of resveratrol production decreased gradually with increasing linker length. In silico analysis showed that the prediction of homology models of fusion proteins was consistent with the in vitro and in vivo results.

journal_name

Mol Biosyst

journal_title

Molecular bioSystems

authors

Guo H,Yang Y,Xue F,Zhang H,Huang T,Liu W,Liu H,Zhang F,Yang M,Liu C,Lu H,Zhang Y,Ma L

doi

10.1039/c6mb00563b

subject

Has Abstract

pub_date

2017-02-28 00:00:00

pages

598-606

issue

3

eissn

1742-206X

issn

1742-2051

journal_volume

13

pub_type

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