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An engineered yeast efficiently secreting penicillin.

Loknath Gidijala1, Jan A K W Kiel, Rutger D Douma

  • 1Molecular Cell Biology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Haren, The Netherlands.

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Summary

Researchers engineered the yeast Hansenula polymorpha to produce penicillin (PEN), a beta-lactam antibiotic, using genes from Penicillium chrysogenum. This demonstrates functional expression of non-ribosomal peptide synthetases in yeast for pharmaceutical production.

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Area of Science:

  • Biotechnology
  • Microbiology
  • Synthetic Biology

Background:

  • Industrial penicillin (PEN) production relies on Penicillium chrysogenum.
  • Hansenula polymorpha is a yeast with a history of pharmaceutical production.
  • Developing alternative hosts can improve antibiotic manufacturing.

Purpose of the Study:

  • To establish yeast Hansenula polymorpha as a viable host for penicillin production.
  • To explore the functional expression of key penicillin biosynthesis genes in a heterologous yeast system.
  • To investigate the role of peroxisomal localization in efficient penicillin production.

Main Methods:

  • Genetic engineering of H. polymorpha to express Penicillium chrysogenum genes.
  • Co-expression of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS) and Bacillus subtilis sfp gene.
  • Introduction of isopenicillin N synthase, isopenicillin N acyl transferase (IAT), and phenylacetyl CoA ligase (PCL) genes.
  • Comparison of PEN production in wild-type and peroxisome-deficient yeast strains.

Main Results:

  • Functional expression of active ACVS enzyme in H. polymorpha.
  • Production and secretion of biologically active penicillin (PEN) by engineered yeast.
  • PEN production levels comparable to the native P. chrysogenum strain (approx. 1 mg/L).
  • Significantly reduced PEN yield in yeast lacking peroxisomes, highlighting the importance of IAT and PCL localization.

Conclusions:

  • Hansenula polymorpha can be successfully engineered for penicillin production.
  • This study represents the first functional expression of a non-ribosomal peptide synthetase in yeast.
  • The findings open avenues for yeast-based production of diverse peptides and beta-lactam antibiotics.
  • Peroxisomal localization of specific enzymes is crucial for optimal penicillin yield.