A temperature-induced chitosanase bacterial cell-surface display system for the efficient production of chitooligosaccharides
| Autor: | Shuo Zhang, Shaofeng Rong, Shimin Guan, Yangzhi Ye, Baoguo Cai, Qianqian Li, Tuantuan Wang |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Glycoside Hydrolases Recombinant Fusion Proteins Oligosaccharides Bioengineering Chitobiose medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology Chitosan Gel permeation chromatography 03 medical and health sciences chemistry.chemical_compound Hydrolysis Bacterial Proteins 010608 biotechnology Enzymatic hydrolysis medicine Escherichia coli Chitosanase chemistry.chemical_classification Chromatography Temperature General Medicine 030104 developmental biology Enzyme chemistry Cell Surface Display Techniques Biotechnology Bacterial Outer Membrane Proteins |
| Zdroj: | Biotechnology letters. 43(8) |
| ISSN: | 1573-6776 |
| Popis: | To establish a temperature-induced chitosanase bacterial cell-surface display system to produce chitooligosaccharides (COSs) efficiently for industrial applications. Temperature-inducible chitosanase CSN46A bacterial surface display systems containing one or two copies of ice nucleation protein (InaQ-N) as anchoring motifs were successfully constructed on the basis of Escherichia coli and named as InaQ-N-CSN46A (1 copy) and 2InaQ-N-CSN46A (2 copies). The specific enzyme activity of 2InaQ-N-CSN46A reached 761.34 ± 0.78 U/g cell dry weight, which was 45.6% higher than that of InaQ-N-CSN46A. However, few proteins were detected in the 2InaQ-N-CSN46A hydrolysis system. Therefore, 2InaQ-N-CSN46A had higher hydrolysis efficiency and stability than InaQ-N-CSN46A. Gel permeation chromatography revealed that under the optimum enzymatic hydrolysis temperature, the final products were mainly chitobiose and chitotriose. Chitopentaose accumulated (77.62%) when the hydrolysis temperature reached 60 °C. FTIR and NMR analysis demonstrated that the structures of the two hydrolysis products were consistent with those of COSs. In this study, chitosanase was expressed on the surfaces of E. coli by increasing the induction temperature, and chitosan was hydrolysed directly without enzyme purification steps. This study provides a novel strategy for industrial COS production. |
| Databáze: | OpenAIRE |
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