Recombinant expression, characterization, and application of a phospholipase B from Fusarium oxysporum
| Autor: | Yongmei Xia, Dening Ji, Lingang Yu, Jing Wu, Lingqia Su, Tao Xiumei |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Cell Count Bioengineering Protein Engineering 01 natural sciences Applied Microbiology and Biotechnology Pichia law.invention Pichia pastoris Gene product 03 medical and health sciences Bioreactors Bacterial Proteins Fusarium law 010608 biotechnology Enzyme Stability Fusarium oxysporum Bioreactor Cloning Molecular Lipase chemistry.chemical_classification Phospholipase B biology business.industry General Medicine biology.organism_classification Recombinant Proteins Biotechnology Enzyme Activation Petroleum 030104 developmental biology Enzyme chemistry Biochemistry Batch Cell Culture Techniques biology.protein Recombinant DNA Lysophospholipase business |
| Zdroj: | Journal of Biotechnology. 242:92-100 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/j.jbiotec.2016.12.009 |
| Popis: | In this study, a gene encoding a putative lipase from Fusarium oxysporum was optimized via codon optimization and expressed in Pichia pastoris KM71. The gene product was identified as a phospholipase B (PLB). The engineered P. pastoris was further cultured in a 3.6-L bioreactor. After optimization of the induction conditions, this system produced 6.6 mg mL−1 protein and 6503.8 U mL−1 PLB activity in the culture medium. Efficient expression of this PLB in P. pastoris should reduce the costs of production and application. The purified enzyme, with a specific activity of 1170 U mg−1, was optimally active at pH 5.0 and 55 °C. The results of a degumming experiment performed using the recombinant PLB showed that the phosphorus content of a test oil was decreased from 75.88 ppm to 3.3 ppm in 2 h under optimal reaction conditions. This study provides a basis for the industrial use of F. oxysporum PLB in oil degumming applications. |
| Databáze: | OpenAIRE |
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