Complete Genome Sequences of Four Natural
            <i>Pseudomonas</i>
            Isolates That Catabolize a Wide Range of Aromatic Compounds Relevant to Lignin Valorization

Hatmaker, E. Anne; Presley, Gerald N.; Cannon, Olivia  N.; Guss, Adam M.; Elkins, James G.

doi:10.1128/mra.00975-20

Cited by 1 publication

(1 citation statement)

References 20 publications

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“…Isolated genomic DNA (~5 μg) was sent to DOE Joint Genome Institute for PacBio Sequencing. Integration of pW17 was verified by the PacBio RSII system (Pacific Biosciences) with default settings and parameters (Hatmaker et al, 2020 ; Wang et al, 2019 ). PacBio sequencing confirmed the integration locus in KT2440 (PPK706, clones 1–4) but integration locus could not be confirmed for M2 (PP2M729, clones 1–4) due to multiple sequence alignments to pW17.…”

Section: Methodsmentioning

confidence: 99%

Development of genetic tools for heterologous protein expression in a pentose‐utilizing environmental isolate of Pseudomonas putida

Gauttam

Eng

Zhao

et al. 2023

Microbial Biotechnology

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Pseudomonas putida has emerged as a promising host for the conversion of biomass-derived sugars and aromatic intermediates into commercially relevant biofuels and bioproducts. Most of the strain development studies previously published have focused on P. putida KT2440, which has been engineered to produce a variety of non-native bioproducts. However, P. putida is not capable of metabolizing pentose sugars, which can constitute up to 25% of biomass hydrolysates. Related P. putida isolates that metabolize a larger fraction of biomass-derived carbon may be attractive as complementary hosts to P. putida KT2440. Here we describe genetic tool development for P. putida M2, a soil isolate that can metabolize pentose sugars. The functionality of five inducible promoter systems and 12 ribosome binding sites was assessed to regulate gene expression. The utility of these expression systems was confirmed by the production of indigoidine from C6 and C5 sugars. Chromosomal integration and expression of non-native genes was achieved by using chassis-independent recombinase-assisted genome engineering (CRAGE) for single-step gene integration of biosynthetic pathways directly into the genome of P. putida M2. These genetic tools provide a foundation to develop hosts complementary to P. putida KT2440 and expand the ability of this versatile microbial group to convert biomass to bioproducts. − , m k + ) supE44 thi −1 gyrA996 relA1 phoA Hanahan (1983) E. coli BW29427 A conjugal donor strain to transfer plasmids, auxotrophic to diaminopimelic acid Wang et al. (2019) P. putida KT2440 Wild type ATCC 12633 P. putida M2 Accession number JADOUD010000001 Park, Fong, et al. (2022) P. putida PPK1 (PP1; JPUB_014489) P. putida KT2440 carrying pRGPDuo2; Kan R Gauttam et al. (2020) P. putida PPK2 (PP2; JPUB_014491) P. putida KT2440 carrying pRGPDuo2-sfGFPtac RBS0; Kan R Gauttam et al. (2020) P. putida PPK3 (PP3; JPUB_014493) P. putida KT2440 carrying pRGPDuo2-sfGFPtet RBS0; Kan R Gauttam et al. (2020) P. putida PPK7 (PP7; JPUB_014775) P. putida KT2440 carrying pRGPDuo1; Gent R Gauttam et al. (2021) P. putida PPK9 (PP9; JPUB_014779) P. putida KT2440 carrying pRGPDuo1-sfGFPtac RBS0; Gent R Gauttam et al. (2021) P. putida PPK12 (PP12; JPUB_014785) P. putida KT2440 carrying pRGPDuo3; Gent R Gauttam et al. (2021) P. putida PPK13 (PP13; JPUB_014787) P. putida KT2440 carrying pRGPDuo4; Kan R Gauttam et al. (2021) P. putida PPK14 (PP14; JPUB_014810) P. putida KT2440 carrying pRGPDuo3-sfGFPbad RBS0; Gent R Gauttam et al. (2021) P. putida PPK16 (PP16; JPUB_014791) P. putida KT2440 carrying pRGPDuo4-sfGFPbad RBS0; Kan R Gauttam et al. (2021) P. putida PP2M226 (JPUB_020213) P. putida M2 carrying pRGPDuo1; Gent R This study P. putida PP2M227 (JPUB_020214) P. putida M2 carrying pRGPDuo2; Kan R This study P. putida PP2M229 (JPUB_020215) P. putida M2 carrying pRGPDuo1-sfGFPtac RBS0; Gent R This study P. putida PP2M230 P. putida M2 carrying pRGPDuo2-sfGFPtet RBS0; Kan R This study P. putida PP2M231 (JPUB_020216) P. putida M2 carrying pRGPDuo2-sfGFPtac RBS0; Kan R ...

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Section: Methodsmentioning

confidence: 99%