N-Terminal Fused Signal Peptide Prompted Extracellular Production of a Bacillus-Derived Alkaline and Thermo Stable Xylanase in E. coli Through Cell Autolysis
| Autor: | Fenghong Huang, Huahua He, Liang Feng, Li Yi, Chan Yu, Haoran Ge, Ting Deng, Faying Zhang |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Signal peptide Autolysis (biology) Lysis Bacillus Bioengineering Protein Sorting Signals 01 natural sciences Applied Microbiology and Biotechnology Biochemistry 010608 biotechnology Enzyme Stability Escherichia coli Extracellular Molecular Biology Endo-1 4-beta Xylanases biology 010405 organic chemistry Bacillus pumilus Chemistry Hydrolysis Cell autolysis Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification 0104 chemical sciences Periplasm Xylanase Extracellular Space Bacterial outer membrane Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 192:339-352 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | Xylanases are extensively used as industrial enzymes for its ability of hydrolyzing xylan to oligosaccharides. Here, XynHB, a thermo and alkaline stable xylanase derived from Bacillus pumilus HBP8, was extracellularly produced in E. coli cells through N-terminal-fused signal peptides. We found that the matured XynHB itself could be auto-secreted out of E. coli BL21(DE3) cells at a very low level, and two Sec-pathway signal peptides, PelB and OmpA, and one dual Sec-Tat-pathway signal peptide, FhuD, could effectively prompt its extracellular production up to 12-fold. Our results showed that PelB signal peptide led to the highest extracellular production of XynHB for approximately 54.1 μg/mL, and FhuD-fused XynHB possessed the highest specific activity of 1746.0 U/mg at 70 °C. Meanwhile, our studies also indicated that PelB- and FhuD-fused XynHB might disrupt E. coli cells' periplasm during their secretion process, thus causing cell lysis to facilitate their extracellular production. Moreover, further characterization revealed that the extracellular production of XynHB was not affected by the outer membrane permeability of E. coli cells. Our studies provided an advantageous strategy for the extracellular production of xylanase in E. coli, which may also be used for E. coli autolysis in the future. |
| Databáze: | OpenAIRE |
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