Expression and Optimization Thermo-Stable Lysine Aminopeptidase in P. pastoris

doi:10.3969/j.issn.1673-1689.2018.06.004

Home > Archive>Volume 37, Issue 6, 2018 >589-594. DOI:10.3969/j.issn.1673-1689.2018.06.004

Expression and Optimization Thermo-Stable Lysine Aminopeptidase in P. pastoris
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                        10.3969/j.issn.1673-1689.2018.06.004
                    
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The aim of this study was toachieve heterologous expression of thermo-Stable Lysine Aminopeptidase（PLAP），the PLAP gene thathave been investigated producing a thermo-stable Lysine Aminopeptidasewas cloned from the genomic DNA of Pseudomonas aeruginosa NJ-814 by PCR.The expression vector pPIC9K-PLAP was constructed to express extracellular protein in P. pastoris GS115. After preliminary screening of the recombinant P. pastoris，the optimized fermentation conditions based on single factor experiment wereidentified as follows：induction methanol concentration was 15 g/L，induction pH was 5.0，induction temperature was 28 ℃，the medium volume was 25 mL/250 mL，the amount of Co2+ was 50 μmol/mL，induction sorbitol was 9 g/L. Finally，the enzyme activity was 5.2 times as in original conditions，the specific activitywas 2.74 U/mg. The result showed that PLAP gene has been successfully transferred and expressed in the Pichia pastoris GS115。

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LIU Qiang, TIAN Yaping. Expression and Optimization Thermo-Stable Lysine Aminopeptidase in P. pastoris[J]. Journal of Food Science and Biotechnology,2018,37(6):589-594.

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Online: September 27,2018
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