|
| Heterologous expression of linoleate isomerase from Propionibacterium acnes in |
| |
| DOI： |
| KeyWord:Pichia pastoris linoleate isomerase heterologous expression conjugated linoleic acid resting cell catalysis |
| FundProject:国家自然科学基金优秀青年基金（31722041） |
|
| Hits: 1131 |
| Download times: 735 |
| Abstract: |
| Linoleate acid isomerase (PAI) from Propionibacterium acnes was a linoleate acid isomerase that could generate conjugated linoleic acid (t10, c12-CLA) through single enzyme catalysis. In order to realize the efficient expression of PAI, pPink was used as the expression vector and Pichia pastoris (PichiaPinkTM Strain2) was used as the host strain to construct a Pichia pastoris recombinant strain (Pichia-pPink-His-opai). Finally, the heterologous expression of PAI in Pichia pastoris was realized. The Pichia pastoris recombinant strain was initially screened to obtain the transformant with the highest protein expression. The single factor experiment was used to obtain the best induction conditions for the Pichia pastoris recombinant strain as follows: volume fraction of the inducer methanol 2%, and induction time 24 h. Under these induction conditions, the Pichia pastoris recombinant strain was made into a resting cell catalyst to catalyze the synthsis of t10, c12-CLA. Under the conditions of 28 ℃, 200 r/min, mass concentration of linoleic acid 4 g/L, and reaction time 6 h, the productivity of t10, c12-CLA was 2.12 g/L, and the conversion rate reached 53%. |
| View full text View/Add Comment Download reader |
| Close |
|
|
|
