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3 Biotech

, 9:33 | Cite as

High-level expression of Thermomyces dupontii thermo-alkaline lipase in Pichia pastoris under the control of different promoters

  • Jianrong Wang
  • Tianyu Zhang
  • Yangyuan Li
  • Lilang Li
  • Yong Wang
  • Bo Yang
  • Yonghua WangEmail author
Original Article
  • 31 Downloads

Abstract

In this study, 15 methanol-inducible and 9 constitutive promoters were used to drive the expression of Thermomyces dupontii lipase (TDL) in Pichia pastoris. Of the 15 methanol-inducible promoters, formaldehyde dehydrogenase promoter (PFLD1) showed the highest efficiency in driving lipase production, followed by alcohol oxidase 1 (PAOX1) and dihydroxyacetone synthase (PDAS1) promoters. The maximum lipase activity of transformants with PFLD1, PAOX1 and PDAS1 promoters in 5-l bioreactor was 27,076, 24,159 and 22,342 U/ml, respectively. For the nine constitutive promoters, glycosyl phosphatidyl inositol-anchored protein promoter (PGCW14) produced the highest amount of lipases in a medium containing glucose or glycerol as the only carbon source, followed by mitochondrial alcohol dehydrogenase isozyme (P0472) and glyceraldehyde-3-phosphate dehydrogenase (PGAP) promoters. The maximum lipase yields in 5-l bioreactors under the control of PGCW14, P0472 and PGAP promoters were 17,353, 15,046 and 14,276 U/ml, respectively. The result of this study not only identifies a few highly efficient promoters for the heterologous expression of TDL in P. pastoris, but also casts some insight into the optimization of protein production in heterologous systems.

Keywords

Lipase Pichia pastoris Promoter Thermomyces dupontii 

Notes

Acknowledgements

This work was supported by the National High Technology Project of People’s Republic of China (No. 2014AA093514).

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.

Supplementary material

13205_2018_1531_MOESM1_ESM.docx (10.8 mb)
Supplementary material 1 (DOCX 11069 KB)

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Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  • Jianrong Wang
    • 1
  • Tianyu Zhang
    • 1
  • Yangyuan Li
    • 3
  • Lilang Li
    • 3
  • Yong Wang
    • 3
  • Bo Yang
    • 1
  • Yonghua Wang
    • 2
    Email author
  1. 1.School of Bioscience and BioengineeringSouth China University of TechnologyGuangzhouPeople’s Republic of China
  2. 2.School of Food Science and EngineeringSouth China University of TechnologyGuangzhouPeople’s Republic of China
  3. 3.Guangdong VTR Bio-Tech Co., LtdZhuhaiPeople’s Republic of China

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