Reprogramming<i>Hansenula polymorpha</i>for penicillin production: expression of the<i>Penicillium chrysogenum pcl</i>gene

Gidijala, Loknath; Klei, Ida J. van der; Veenhuis, Marten; Kiel, Jan A.K.W.

doi:10.1111/j.1567-1364.2007.00228.x

Cited by 22 publications

(17 citation statements)

References 36 publications

(41 reference statements)

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“…3). We previously showed that both heterologous proteins are properly synthesized and sorted in H. polymorpha [8], [9]. To test whether HpPen4 cells produced and secreted the β-lactam antibiotic penicillin G (PenG), cells were grown in media supplemented with AAA and phenylacetic acid (PAA), the PenG side chain precursor.…”

Section: Resultsmentioning

confidence: 99%

An Engineered Yeast Efficiently Secreting Penicillin

et al. 2009

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This study aimed at developing an alternative host for the production of penicillin (PEN). As yet, the industrial production of this β-lactam antibiotic is confined to the filamentous fungus Penicillium chrysogenum. As such, the yeast Hansenula polymorpha, a recognized producer of pharmaceuticals, represents an attractive alternative. Introduction of the P. chrysogenum gene encoding the non-ribosomal peptide synthetase (NRPS) δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS) in H. polymorpha, resulted in the production of active ACVS enzyme, when co-expressed with the Bacillus subtilis sfp gene encoding a phosphopantetheinyl transferase that activated ACVS. This represents the first example of the functional expression of a non-ribosomal peptide synthetase in yeast. Co-expression with the P. chrysogenum genes encoding the cytosolic enzyme isopenicillin N synthase as well as the two peroxisomal enzymes isopenicillin N acyl transferase (IAT) and phenylacetyl CoA ligase (PCL) resulted in production of biologically active PEN, which was efficiently secreted. The amount of secreted PEN was similar to that produced by the original P. chrysogenum NRRL1951 strain (approx. 1 mg/L). PEN production was decreased over two-fold in a yeast strain lacking peroxisomes, indicating that the peroxisomal localization of IAT and PCL is important for efficient PEN production. The breakthroughs of this work enable exploration of new yeast-based cell factories for the production of (novel) β-lactam antibiotics as well as other natural and semi-synthetic peptides (e.g. immunosuppressive and cytostatic agents), whose production involves NRPS's.

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Section: Resultsmentioning

confidence: 99%

An Engineered Yeast Efficiently Secreting Penicillin

et al. 2009

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“…In P. chrysogenum, microbodies (peroxisomes) are essential for penicillin biosynthesis because the two final enzymatic steps catalyzed by acyl-CoA:isopenicillinN acyltransferase 28 and phenylacetyl-CoA ligase 29 are located in these organelles. Moreover, high-producing strains have enhanced microbody volume fractions (Fig.…”

Section: Microbodiesmentioning

confidence: 99%

Genome sequencing and analysis of the filamentous fungus Penicillium chrysogenum

et al. 2008

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“…Microbodies are organelles involved in the final steps of the penicillin pathway, which are catalyzed by the acyl-coenzyme A: isopenicillin N acyltransferase (IAT) and phenylacetyl-CoA ligase [18, 55, 56]. In fact, it has been reported that microbody volume fractions are enhanced in high-producing strains [57].…”

Section: Classical Strain Improvement and Molecular Genomic-transcmentioning

confidence: 99%

Proteomics Shows New Faces for the Old Penicillin ProducerPenicillium chrysogenum

Barreiro

Martı́n

Garcı́a-Estrada

2012

Journal of Biomedicine and Biotechnology

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Fungi comprise a vast group of microorganisms including the Ascomycota (majority of all described fungi), the Basidiomycota (mushrooms or higher fungi), and the Zygomycota and Chytridiomycota (basal or lower fungi) that produce industrially interesting secondary metabolites, such as β-lactam antibiotics. These compounds are one of the most commonly prescribed drugs world-wide. Since Fleming's initial discovery ofPenicillium notatum80 years ago, the role ofPenicilliumas an antimicrobial source became patent. After the isolation ofPenicillium chrysogenumNRRL 1951 six decades ago, classical mutagenesis and screening programs led to the development of industrial strains with increased productivity (at least three orders of magnitude). The new “omics” era has provided the key to understand the underlying mechanisms of the industrial strain improvement process. The review of different proteomics methods applied toP. chrysogenumhas revealed that industrial modification of this microorganism was a consequence of a careful rebalancing of several metabolic pathways. In addition, the secretome analysis ofP. chrysogenumhas opened the door to new industrial applications for this versatile filamentous fungus.

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ReprogrammingHansenula polymorphafor penicillin production: expression of thePenicillium chrysogenum pclgene

Cited by 22 publications

References 36 publications

An Engineered Yeast Efficiently Secreting Penicillin

An Engineered Yeast Efficiently Secreting Penicillin

Genome sequencing and analysis of the filamentous fungus Penicillium chrysogenum

Proteomics Shows New Faces for the Old Penicillin ProducerPenicillium chrysogenum

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