Advanced pathway engineering for phototrophic putrescine production

Freudenberg R, Wittemeier L, Einhaus A, Baier T, Kruse O (2022)
Plant Biotechnology Journal.

Zeitschriftenaufsatz | E-Veröff. vor dem Druck | Englisch
 
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Abstract / Bemerkung
The polyamine putrescine (1,4-diaminobutane) contributes to cellular fitness in most organisms, where it is derived from the amino acids ornithine or arginine. In the chemical industry, putrescine serves as a versatile building block for polyamide synthesis. The green microalga Chlamydomonas reinhardtii accumulates relatively high putrescine amounts, which, together with recent advances in genetic engineering, enables the generation of a powerful green cell factory to promote sustainable biotechnology for base chemical production. Here, we report a systematic investigation of the native putrescine metabolism in C. reinhardtii, leading to the first CO2-based bio-production of putrescine, by employing modern synthetic biology and metabolic engineering strategies. A CRISPR/Cas9-based knockout of key enzymes of the polyamine biosynthesis pathway identified ornithine decarboxylase 1 (ODC1) as a gatekeeper for putrescine accumulation and demonstrated that the arginine decarboxylase (ADC) route is likely inactive and that amine oxidase 2 (AMX2) is mainly responsible for putrescine degradation in C. reinhardtii. A 4.5-fold increase of cellular putrescine levels was achieved by engineered overexpression of potent candidate ornithine decarboxylases (ODCs). We identified unexpected substrate promiscuity in two bacterial ODCs, which exhibited co-production of cadaverine and 4-aminobutanol. Final pathway engineering included overexpression of recombinant arginases for improved substrate availability as well as functional knockout of putrescine degradation, which resulted in a 10-fold increase in cellular putrescine titers and yielded 200 mg/L in phototrophic high cell density cultivations after 10 days.
Erscheinungsjahr
2022
Zeitschriftentitel
Plant Biotechnology Journal
ISSN
1467-7644
eISSN
1467-7652
Page URI
https://pub.uni-bielefeld.de/record/2964020

Zitieren

Freudenberg R, Wittemeier L, Einhaus A, Baier T, Kruse O. Advanced pathway engineering for phototrophic putrescine production. Plant Biotechnology Journal. 2022.
Freudenberg, R., Wittemeier, L., Einhaus, A., Baier, T., & Kruse, O. (2022). Advanced pathway engineering for phototrophic putrescine production. Plant Biotechnology Journal. https://doi.org/10.1111/pbi.13879
Freudenberg, R., Wittemeier, L., Einhaus, A., Baier, T., and Kruse, O. (2022). Advanced pathway engineering for phototrophic putrescine production. Plant Biotechnology Journal.
Freudenberg, R., et al., 2022. Advanced pathway engineering for phototrophic putrescine production. Plant Biotechnology Journal.
R. Freudenberg, et al., “Advanced pathway engineering for phototrophic putrescine production”, Plant Biotechnology Journal, 2022.
Freudenberg, R., Wittemeier, L., Einhaus, A., Baier, T., Kruse, O.: Advanced pathway engineering for phototrophic putrescine production. Plant Biotechnology Journal. (2022).
Freudenberg, Robert, Wittemeier, Luisa, Einhaus, Alexander, Baier, Thomas, and Kruse, Olaf. “Advanced pathway engineering for phototrophic putrescine production”. Plant Biotechnology Journal (2022).

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PMID: 35748533
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