Dehydrogenation Mechanism of Three Stereoisomers of Butane-2,3-Diol in Pseudomonas putida KT2440

Pseudomonas putida KT2440 is a promising chassis of industrial biotechnology due to its metabolic versatility. Butane-2,3-diol (2,3-BDO) is a precursor of numerous value-added chemicals. It is also a microbial metabolite which widely exists in various habiting environments of P. putida KT2440. It was reported that P. putida KT2440 is able to use 2,3-BDO as a sole carbon source for growth. There are three stereoisomeric forms of 2,3-BDO: (2R,3R)-2,3-BDO, meso-2,3-BDO and (2S,3S)-2,3-BDO. However, whether P. putida KT2440 can utilize three stereoisomeric forms of 2,3-BDO has not been elucidated. Here, we revealed the genomic and enzymic basis of P. putida KT2440 for dehydrogenation of different stereoisomers of 2,3-BDO into acetoin, which will be channeled to central mechanism via acetoin dehydrogenase enzyme system. (2R,3R)-2,3-BDO dehydrogenase (PP0552) was detailedly characterized and identified to participate in (2R,3R)-2,3-BDO and meso-2,3-BDO dehydrogenation. Two quinoprotein alcohol dehydrogenases, PedE (PP2674) and PedH (PP2679), were confirmed to be responsible for (2S,3S)-2,3-BDO dehydrogenation. The function redundancy and inverse regulation of PedH and PedE by lanthanide availability provides a mechanism for the adaption of P. putida KT2440 to variable environmental conditions. Elucidation of the mechanism of 2,3-BDO catabolism in P. putida KT2440 would provide new insights for bioproduction of 2,3-BDO-derived chemicals based on this robust chassis.

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Outer membrane vesicles catabolize lignin-derived aromatic compounds in Pseudomonas putida KT2440

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1921073117 ◽

2020 ◽

Vol 117 (17) ◽

pp. 9302-9310 ◽

Cited By ~ 7

Author(s):

Davinia Salvachúa ◽

Allison Z. Werner ◽

Isabel Pardo ◽

Martyna Michalska ◽

Brenna A. Black ◽

...

Keyword(s):

Pseudomonas Putida ◽

Outer Membrane ◽

Aromatic Compounds ◽

Value Added ◽

Membrane Vesicles ◽

Outer Membrane Vesicles ◽

Pseudomonas Putida Kt2440 ◽

Rich Media

Lignin is an abundant and recalcitrant component of plant cell walls. While lignin degradation in nature is typically attributed to fungi, growing evidence suggests that bacteria also catabolize this complex biopolymer. However, the spatiotemporal mechanisms for lignin catabolism remain unclear. Improved understanding of this biological process would aid in our collective knowledge of both carbon cycling and microbial strategies to valorize lignin to value-added compounds. Here, we examine lignin modifications and the exoproteome of three aromatic–catabolic bacteria: Pseudomonas putida KT2440, Rhodoccocus jostii RHA1, and Amycolatopsis sp. ATCC 39116. P. putida cultivation in lignin-rich media is characterized by an abundant exoproteome that is dynamically and selectively packaged into outer membrane vesicles (OMVs). Interestingly, many enzymes known to exhibit activity toward lignin-derived aromatic compounds are enriched in OMVs from early to late stationary phase, corresponding to the shift from bioavailable carbon to oligomeric lignin as a carbon source. In vivo and in vitro experiments demonstrate that enzymes contained in the OMVs are active and catabolize aromatic compounds. Taken together, this work supports OMV-mediated catabolism of lignin-derived aromatic compounds as an extracellular strategy for nutrient acquisition by soil bacteria and suggests that OMVs could potentially be useful tools for synthetic biology and biotechnological applications.

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Biochemical and Molecular Characterization of theBacillus subtilis Acetoin Catabolic Pathway

Journal of Bacteriology ◽

10.1128/jb.181.12.3837-3841.1999 ◽

1999 ◽

Vol 181 (12) ◽

pp. 3837-3841 ◽

Cited By ~ 62

Author(s):

Min Huang ◽

Fred Bernd Oppermann-Sanio ◽

Alexander Steinbüchel

Keyword(s):

Bacillus Subtilis ◽

Carbon Source ◽

Sole Carbon Source ◽

Enzyme System ◽

Gene Clusters ◽

Catabolic Pathway ◽

E Coli ◽

Homologous Genes ◽

Dehydrogenase Enzyme

ABSTRACT A recent study indicated that Bacillus subtiliscatabolizes acetoin by enzymes encoded by the acu gene cluster (F. J. Grundy, D. A. Waters, T. Y. Takova, and T. M. Henkin, Mol. Microbiol. 10:259–271, 1993) that are completely different from those in the multicomponent acetoin dehydrogenase enzyme system (AoDH ES) encoded by aco gene clusters found before in all other bacteria capable of utilizing acetoin as the sole carbon source for growth. By hybridization with a DNA probe covering acoA and acoB of the AoDH ES from Clostridium magnum, genomic fragments from B. subtilis harboring acoA, acoB,acoC, acoL, and acoR homologous genes were identified, and some of them were functionally expressed inE. coli. Furthermore, acoA was inactivated inB. subtilis by disruptive mutagenesis; these mutants were impaired to express PPi-dependent AoDH E1 activity to remove acetoin from the medium and to grow with acetoin as the carbon source. Therefore, acetoin is catabolized in B. subtilis by the same mechanism as all other bacteria investigated so far, leaving the function of the previously described acu genes obscure.

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Generation of ionic liquid tolerant Pseudomonas putida KT2440 strains via adaptive laboratory evolution

Green Chemistry ◽

10.1039/d0gc01663b ◽

2020 ◽

Vol 22 (17) ◽

pp. 5677-5690 ◽

Cited By ~ 1

Author(s):

Hyun Gyu Lim ◽

Bonnie Fong ◽

Geovanni Alarcon ◽

Harsha D. Magurudeniya ◽

Thomas Eng ◽

...

Keyword(s):

Ionic Liquid ◽

Ionic Liquids ◽

Pseudomonas Putida ◽

Low Cost ◽

Industrial Biotechnology ◽

Adaptive Laboratory Evolution ◽

Pseudomonas Putida Kt2440 ◽

Laboratory Evolution

Pseudomonas putida KT2440, a promising microbial platform for industrial biotechnology was tolerized to low-cost biomass decomposing ionic liquids via the adaptive laboratory evolution.

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