Enhanced chaotrope tolerance and (S)-2-hydroxypropiophenone production by recombinant Pseudomonas putida engineered with Pprl from Deinococcus radiodurans.

Autor: Kordesedehi R; Department of Biotechnology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran., Shahpiri A; Department of Biotechnology, College of Agriculture, Isfahan University of Technology, Isfahan, Iran., Asadollahi MA; Department of Biotechnology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran., Biria D; Department of Biotechnology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran., Nikel PI; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark.
Jazyk: angličtina
Zdroj: Microbial biotechnology [Microb Biotechnol] 2024 Mar; Vol. 17 (3), pp. e14448.
DOI: 10.1111/1751-7915.14448
Abstrakt: Pseudomonas putida is a soil bacterium with multiple uses in fermentation and biotransformation processes. P. putida ATCC 12633 can biotransform benzaldehyde and other aldehydes into valuable α-hydroxyketones, such as (S)-2-hydroxypropiophenone. However, poor tolerance of this strain toward chaotropic aldehydes hampers efficient biotransformation processes. To circumvent this problem, we expressed the gene encoding the global regulator PprI from Deinococcus radiodurans, an inducer of pleiotropic proteins promoting DNA repair, in P. putida. Fine-tuned gene expression was achieved using an expression plasmid under the control of the LacI Q /P trc system, and the cross-protective role of PprI was assessed against multiple stress treatments. Moreover, the stress-tolerant P. putida strain was tested for 2-hydroxypropiophenone production using whole resting cells in the presence of relevant aldehyde substrates. P. putida cells harbouring the global transcriptional regulator exhibited high tolerance toward benzaldehyde, acetaldehyde, ethanol, butanol, NaCl, H 2 O 2 and thermal stress, thereby reflecting the multistress protection profile conferred by PprI. Additionally, the engineered cells converted aldehydes to 2-hydroxypropiophenone more efficiently than the parental P. putida strain. 2-Hydroxypropiophenone concentration reached 1.6 g L -1 upon a 3-h incubation under optimized conditions, at a cell concentration of 0.033 g wet cell weight mL -1 in the presence of 20 mM benzaldehyde and 600 mM acetaldehyde. Product yield and productivity were 0.74 g 2-HPP g -1 benzaldehyde and 0.089 g 2-HPP g cell dry weight -1  h -1 , respectively, 35% higher than the control experiments. Taken together, these results demonstrate that introducing PprI from D. radiodurans enhances chaotrope tolerance and 2-HPP production in P. putida ATCC 12633.
(© 2024 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd.)
Databáze: MEDLINE
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