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Pentapeptide prosequence enhances expression and structure folding of recombinant Thermomyces lanuginosus lipase in Pichia pastoris.

Abstract
Propeptides have been demonstrated to influence many properties of their mature proteins as intramolecular chaperones. In our study, the role of pentapeptide prosequence of Thermomyces lanuginosus lipase (TLL) was determined through its effect on the recombinant expression of TLL in P. pastoris. The result showed that the average lipase activity of recombinant strains GS-pTL reached 434.32 U/mL, higher than that of GS-TL 377.71 U/mL, indicating that the propeptide "SPIRR-" sequence is beneficial for enhancing TLL expression. In addition, rational design and site-directed mutation were performed to increase the propeptide's hydrophobicity to explore its possible function mechanism. The fermentation result of the recombinant strains with modified propeptide showed that the extracellular lipase activity of GS-pTL-VP variant reached 483.29 U/mL, increasing by 11.27% compared with that of GS-pTL (434.32 U/mL). Therefore, the increase of hydrophobicity in propeptide was considered to be advantageous to enhance the expression of lipase TLL, possibly through assisting the protein nucleation as a hydrophobic scaffold. This study would provide new insight into the role of short propeptide in protein expression and structure folding.

PMID: 28641563 [Pubmed - Publisher]

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