title: [Effect of signal peptides on the expression of laccase in Pichia pastoris].
abstract: OBJECTIVE
  Expression level of laccase POXA1c would be increased by screening effective signal peptide in Pichia pastoris.
  
  
  METHODS
  According to 2D-gel and profile of P. pastoris genome sequence, seven signal peptides from high secreted endogenous proteins of P. pastoris X-33 were chosen to evaluate their secreted ability by using POXA1c as reporter protein.
  
  
  RESULTS
  Compared with POXA1c's native signal peptide, the signal peptide of repressible acid phosphatases PHO5 and lectin-like protein FLO10 showed 2. 5-fold and 2-fold increase of laccase activity. Furthermore, PHO5-αpro and FLO10-αpro were constructed by fusing signal peptide of PHO5 and FLO10 with pro-peptide of α-MF respectively. The laccase activity under the leading of FLO10-αpro and PHO5-αpro showed 3-fold and 3. 5- fold laccase activity higher than native signal peptide, and showed 20% and 40% increase compared with saccharomyces cerevisiae α-MF signal respectively.
  
  
  CONCLUSIONS
  Signal peptides from high secreted endogenous proteins of P. pastoris X-33 could be effectively used to lead laccase expression in P. pastoris. The activity of POXA1c under the leading of the PHO5-αpro signal peptide was 57.98 U/mL after high density fermentation.
authors: Yiping
authors: Y
authors: Shi
authors: Xue
authors: X
authors: Zhou
authors: Meirong
authors: M
authors: Hu
authors: Jianhui
authors: J
authors: Lin
authors: Yong
authors: Y
authors: Tao
authors: Jianzhong
authors: J
authors: Huang
mesh_terms: D000135:Acid Phosphatase / Q000502:physiology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D005285:Fermentation; D042845:Laccase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D021382:Protein Sorting Signals / Q000502:physiology*
chemical_list: D021382:Protein Sorting Signals; D042845:Laccase; D000135:Acid Phosphatase