Effect of flexible linker length on the activity of fusion protein 4-coumaroyl-CoA ligase::stilbene synthase
| Autor: | Huang Tiran, Yadong Yang, Heshu Lü, Yansheng Zhang, Mingfeng Yang, Fenqiang Zhang, Lanqing Ma, Guo Huili, Zhang Hong, Liu Wenbin, Liu Huan, Liu Chunmei, Feiyan Xue |
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| Rok vydání: | 2017 |
| Předmět: |
Models
Molecular 0301 basic medicine Protein Conformation Recombinant Fusion Proteins Saccharomyces cerevisiae Arabidopsis Gene Expression Peptide Biology medicine.disease_cause Structure-Activity Relationship 03 medical and health sciences Protein structure In vivo Escherichia coli medicine Molecular Biology chemistry.chemical_classification DNA ligase biology.organism_classification Fusion protein Enzyme Activation Kinetics 030104 developmental biology chemistry Biochemistry Acyl Coenzyme A Linker Acyltransferases Biotechnology |
| Zdroj: | Molecular BioSystems. 13:598-606 |
| ISSN: | 1742-2051 1742-206X |
| DOI: | 10.1039/c6mb00563b |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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