Oligomerization triggered by foldon: a simple method to enhance the catalytic efficiency of lichenase and xylanase
| Autor: | Ge Huihua, Guangya Zhang, Shuyu Wu, Xinzhe Wang, Dandan Zhang |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Protein Folding Glycoside Hydrolases lcsh:Biotechnology Biology Protein Engineering 010402 general chemistry Enzyme engineering 01 natural sciences Catalysis Gene Expression Regulation Enzymologic Substrate Specificity Evolution Molecular Foldon 03 medical and health sciences Hydrolysis chemistry.chemical_compound Protein Domains lcsh:TP248.13-248.65 Escherichia coli Oligomerization Non-chromatographic purification Catalytic efficiency chemistry.chemical_classification Endo-1 4-beta Xylanases Wild type Gene Expression Regulation Bacterial Protein engineering Combinatorial chemistry Elastin-like polypeptides 0104 chemical sciences Enzyme Activation Genetic Enhancement 030104 developmental biology Enzyme Monomer Biochemistry chemistry Xylanase Protein Multimerization Bacillus subtilis Research Article Biotechnology |
| Zdroj: | BMC Biotechnology BMC Biotechnology, Vol 17, Iss 1, Pp 1-10 (2017) |
| ISSN: | 1472-6750 |
| DOI: | 10.1186/s12896-017-0380-3 |
| Popis: | Effective and simple methods that lead to higher enzymatic efficiencies are highly sough. Here we proposed a foldon-triggered trimerization of the target enzymes with significantly improved catalytic performances by fusing a foldon domain at the C-terminus of the enzymes via elastin-like polypeptides (ELPs). The foldon domain comprises 27 residues and can forms trimers with high stability. Lichenase and xylanase can hydrolyze lichenan and xylan to produce value added products and biofuels, and they have great potentials as biotechnological tools in various industrial applications. We took them as the examples and compared the kinetic parameters of the engineered trimeric enzymes to those of the monomeric and wild type ones. When compared with the monomeric ones, the catalytic efficiency (k cat /K m ) of the trimeric lichenase and xylanase increased 4.2- and 3.9- fold. The catalytic constant (k cat ) of the trimeric lichenase and xylanase increased 1.8- fold and 5.0- fold than their corresponding wild-type counterparts. Also, the specific activities of trimeric lichenase and xylanase increased by 149% and 94% than those of the monomeric ones. Besides, the recovery of the lichenase and xylanase activities increased by 12.4% and 6.1% during the purification process using ELPs as the non-chromatographic tag. The possible reason is the foldon domain can reduce the transition temperature of the ELPs. The trimeric lichenase and xylanase induced by foldon have advantages in the catalytic performances. Besides, they were easier to purify with increased purification fold and decreased the loss of activities compared to their corresponding monomeric ones. Trimerizing of the target enzymes triggered by the foldon domain could improve their activities and facilitate the purification, which represents a simple and effective enzyme-engineering tool. It should have exciting potentials both in industrial and laboratory scales. |
| Databáze: | OpenAIRE |
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