Engineered Pseudomonas putida KT2440 co-utilizes galactose and glucose

Peabody, V, George L.; Elmore, Joshua R.; Martinez-Baird, Jessica; Guss, Adam M.

doi:10.1186/s13068-019-1627-0

Engineered Pseudomonas putida KT2440 co-utilizes galactose and glucose

Journal Article · Mon Dec 23 00:00:00 EST 2019 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-019-1627-0· OSTI ID:1618773

Peabody, V, George L.; Elmore, Joshua R.; Martinez-Baird, Jessica;

Abstract Background

Efficient conversion of plant biomass to commodity chemicals is an important challenge that needs to be solved to enable a sustainable bioeconomy. Deconstruction of biomass to sugars and lignin yields a wide variety of low molecular weight carbon substrates that need to be funneled to product. Pseudomonas putida KT2440 has emerged as a potential platform for bioconversion of lignin and the other components of plant biomass. However, P. putida is unable to natively utilize several of the common sugars in hydrolysate streams, including galactose.

Results

In this work, we integrated a De Ley–Doudoroff catabolic pathway for galactose catabolism into the chromosome of P. putida KT2440, using genes from several different organisms. We found that the galactonate catabolic pathway alone (DgoKAD) supported slow growth of P. putida on galactose. Further integration of genes to convert galactose to galactonate and to optimize the transporter expression level resulted in a growth rate of 0.371 h ⁻¹ . Additionally, the best-performing strain was demonstrated to co-utilize galactose with glucose.

Conclusions

We have engineered P. putida to catabolize galactose, which will allow future engineered strains to convert more plant biomass carbon to products of interest. Further, by demonstrating co-utilization of glucose and galactose, continuous bioconversion processes for mixed sugar streams are now possible.

View Journal Article

Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE; USDOE Office of Energy Efficiency and Renewable Energy (EERE)

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1618773

Alternate ID(s):: OSTI ID: 1607137

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Journal Issue: 1 Vol. 12; ISSN 1754-6834

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: Netherlands

Language:: English

References (34)

CRISPR/Cas9-Based Counterselection Boosts Recombineering Efficiency in Pseudomonas putida Aparicio, Tomás; de Lorenzo, Víctor; Martínez-García, Esteban Biotechnology Journal, Vol. 13, Issue 5 https://doi.org/10.1002/biot.201700161	journal	December 2017
Improved performance of Pseudomonas putida in a bioelectrochemical system through overexpression of periplasmic glucose dehydrogenase Yu, Shiqin; Lai, Bin; Plan, Manuel R. Biotechnology and Bioengineering, Vol. 115, Issue 1 https://doi.org/10.1002/bit.26433	journal	October 2017
Lignocellulosic agriculture wastes as biomass feedstocks for second-generation bioethanol production: concepts and recent developments Saini, Jitendra Kumar; Saini, Reetu; Tewari, Lakshmi 3 Biotech, Vol. 5, Issue 4 https://doi.org/10.1007/s13205-014-0246-5	journal	August 2014
The enzymatic transformation of uridine diphosphate glucose into a galactose derivative Leloir, Luis F. Archives of Biochemistry and Biophysics, Vol. 33, Issue 2 https://doi.org/10.1016/0003-9861(51)90096-3	journal	September 1951
D-galactonate utilisation by enteric bacteria The catabolic pathway in Escherichia coli Deacon, Julia; Cooper, R. A. FEBS Letters, Vol. 77, Issue 2 https://doi.org/10.1016/0014-5793(77)80234-2	journal	May 1977
Chemical profiles of switchgrass Hu, Zhoujian; Sykes, Robert; Davis, Mark F. Bioresource Technology, Vol. 101, Issue 9 https://doi.org/10.1016/j.biortech.2009.12.033	journal	May 2010
Innovative Chemicals and Materials from Bacterial Aromatic Catabolic Pathways Johnson, Christopher W.; Salvachúa, Davinia; Rorrer, Nicholas A. Joule, Vol. 3, Issue 6 https://doi.org/10.1016/j.joule.2019.05.011	journal	June 2019
Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440 Linger, Jeffrey G.; Hobdey, Sarah E.; Franden, Mary Ann Metabolic Engineering Communications, Vol. 3 https://doi.org/10.1016/j.meteno.2016.01.005	journal	December 2016
Conversion and assimilation of furfural and 5-(hydroxymethyl)furfural by Pseudomonas putida KT2440 Guarnieri, Michael T.; Ann Franden, Mary; Johnson, Christopher W. Metabolic Engineering Communications, Vol. 4 https://doi.org/10.1016/j.meteno.2017.02.001	journal	June 2017
Development of a high efficiency integration system and promoter library for rapid modification of Pseudomonas putida KT2440 Elmore, Joshua R.; Furches, Anna; Wolff, Gara N. Metabolic Engineering Communications, Vol. 5 https://doi.org/10.1016/j.meteno.2017.04.001	journal	December 2017
Rapid generation of recombinant Pseudomonas putida secondary metabolite producers using yTREX Domröse, Andreas; Weihmann, Robin; Thies, Stephan Synthetic and Systems Biotechnology, Vol. 2, Issue 4 https://doi.org/10.1016/j.synbio.2017.11.001	journal	December 2017
Aromatic catabolic pathway selection for optimal production of pyruvate and lactate from lignin Johnson, Christopher W.; Beckham, Gregg T. Metabolic Engineering, Vol. 28 https://doi.org/10.1016/j.ymben.2015.01.005	journal	March 2015
Pseudomonas putida as a functional chassis for industrial biocatalysis: From native biochemistry to trans-metabolism Nikel, Pablo I.; de Lorenzo, Víctor Metabolic Engineering, Vol. 50 https://doi.org/10.1016/j.ymben.2018.05.005	journal	November 2018
Refactoring the upper sugar metabolism of Pseudomonas putida for co-utilization of cellobiose, xylose, and glucose Dvořák, Pavel; de Lorenzo, Víctor Metabolic Engineering, Vol. 48 https://doi.org/10.1016/j.ymben.2018.05.019	journal	July 2018
Engineering Pseudomonas putida KT2440 for efficient ethylene glycol utilization Franden, Mary Ann; Jayakody, Lahiru N.; Li, Wing-Jin Metabolic Engineering, Vol. 48 https://doi.org/10.1016/j.ymben.2018.06.003	journal	July 2018
Tn7-Based Device for Calibrated Heterologous Gene Expression in Pseudomonas putida Zobel, Sebastian; Benedetti, Ilaria; Eisenbach, Lara ACS Synthetic Biology, Vol. 4, Issue 12 https://doi.org/10.1021/acssynbio.5b00058	journal	July 2015
Holistic bioengineering: rewiring central metabolism for enhanced bioproduction Aslan, Selçuk; Noor, Elad; Bar-Even, Arren Biochemical Journal, Vol. 474, Issue 23 https://doi.org/10.1042/BCJ20170377	journal	November 2017
Pseudomonas putida KT2440 Strain Metabolizes Glucose through a Cycle Formed by Enzymes of the Entner-Doudoroff, Embden-Meyerhof-Parnas, and Pentose Phosphate Pathways Nikel, Pablo I.; Chavarría, Max; Fuhrer, Tobias Journal of Biological Chemistry, Vol. 290, Issue 43 https://doi.org/10.1074/jbc.M115.687749	journal	September 2015
Cloning, Expression, and Characterization of Bacterial L-Arabinose 1-Dehydrogenase Involved in an Alternative Pathway of L-Arabinose Metabolism Watanabe, Seiya; Kodak, Tsutomu; Makino, Keisuke Journal of Biological Chemistry, Vol. 281, Issue 5, p. 2612-2623 https://doi.org/10.1074/jbc.M506477200	journal	December 2005
A Novel α-Ketoglutaric Semialdehyde Dehydrogenase: EVOLUTIONARY INSIGHT INTO AN ALTERNATIVE PATHWAY OF BACTERIAL l-ARABINOSE METABOLISM Watanabe, Seiya; Kodaki, Tsutomu; Makino, Keisuke Journal of Biological Chemistry, Vol. 281, Issue 39 https://doi.org/10.1074/jbc.M602585200	journal	July 2006
Identification and Characterization of l-Arabonate Dehydratase, l-2-Keto-3-deoxyarabonate Dehydratase, and l-Arabinolactonase Involved in an Alternative Pathway of l-Arabinose Metabolism: Novel Evolutionary Insight into Sugar Metabolism Watanabe, Seiya; Shimada, Naoko; Tajima, Kunihiko Journal of Biological Chemistry, Vol. 281, Issue 44, p. 33521-33536 https://doi.org/10.1074/jbc.M606727200	journal	September 2006
Relative substrate affinities of wild-type and mutant forms of the Escherichia coli sugar transporter GalP determined by solid-state NMR Patching, Simon G.; Psakis, Georgios; Baldwin, Stephen A. Molecular Membrane Biology, Vol. 25, Issue 6-7, p. 474-484 https://doi.org/10.1080/09687680802371963	journal	January 2008
Complete nucleotide sequence of Pseudomonas fluortscens D-galactose dehydrogenase gene Sperka, Sabine; Zehelein, Eva; Fiedler, Stefan Nucleic Acids Research, Vol. 17, Issue 13 https://doi.org/10.1093/nar/17.13.5402	journal	January 1989
The Leloir pathway: a mechanistic imperative for three enzymes to change the stereochemical configuration of a single carbon in galactose. Frey, P. A. The FASEB Journal, Vol. 10, Issue 4 https://doi.org/10.1096/fasebj.10.4.8647345	journal	March 1996
The Entner-Doudoroff pathway empowers Pseudomonas putida KT2440 with a high tolerance to oxidative stress : Perturbing the upper metabolism of Chavarría, Max; Nikel, Pablo I.; Pérez-Pantoja, Danilo Environmental Microbiology, Vol. 15, Issue 6 https://doi.org/10.1111/1462-2920.12069	journal	January 2013
Accelerated genome engineering of Pseudomonas putida by I‐ Sce I―mediated recombination and CRISPR ‐Cas9 counterselection Wirth, Nicolas T.; Kozaeva, Ekaterina; Nikel, Pablo I. Microbial Biotechnology, Vol. 13, Issue 1 https://doi.org/10.1111/1751-7915.13396	journal	March 2019
Growth of engineered Pseudomonas putida KT2440 on glucose, xylose, and arabinose: Hemicellulose hydrolysates and their major sugars as sustainable carbon sources Wang, Yan; Horlamus, Felix; Henkel, Marius GCB Bioenergy, Vol. 11, Issue 1 https://doi.org/10.1111/gcbb.12590	journal	January 2019
d-Galactose Dehydrogenase from Pseudomonas fluorescens. Purification, Properties and Structure Blachnitzky, Ernst-Otto; Wengenmayer, Friedrich; Kurz, Gerhart European Journal of Biochemistry, Vol. 47, Issue 2 https://doi.org/10.1111/j.1432-1033.1974.tb03687.x	journal	September 1974
The Metabolic Redox Regime of Pseudomonas putida Tunes Its Evolvability toward Novel Xenobiotic Substrates Akkaya, Özlem; Pérez-Pantoja, Danilo R.; Calles, Belén mBio, Vol. 9, Issue 4 https://doi.org/10.1128/mBio.01512-18	journal	August 2018
Production of medium-chain-length polyhydroxyalkanoates by sequential feeding of xylose and octanoic acid in engineered Pseudomonas putida KT2440 Le Meur, Sylvaine; Zinn, Manfred; Egli, Thomas BMC Biotechnology, Vol. 12, Issue 1 https://doi.org/10.1186/1472-6750-12-53	journal	January 2012
Development of a broad-host-range sacB-based vector for unmarked allelic exchange Marx, Christopher J. BMC Research Notes, Vol. 1, Article No. 1 https://doi.org/10.1186/1756-0500-1-1	journal	January 2008
Genome-Scale Reconstruction and Analysis of the Pseudomonas putida KT2440 Metabolic Network Facilitates Applications in Biotechnology Puchałka, Jacek; Oberhardt, Matthew A.; Godinho, Miguel PLoS Computational Biology, Vol. 4, Issue 10 https://doi.org/10.1371/journal.pcbi.1000210	journal	October 2008
Development of an Optimized Medium, Strain and High-Throughput Culturing Methods for Methylobacterium extorquens Delaney, Nigel F.; Kaczmarek, Maria E.; Ward, Lewis M. PLoS ONE, Vol. 8, Issue 4 https://doi.org/10.1371/journal.pone.0062957	journal	April 2013
Modular Engineering of Biomass Degradation Pathways Chaves, Julie E.; Presley, Gerald N.; Michener, Joshua K. Processes, Vol. 7, Issue 4 https://doi.org/10.3390/pr7040230	journal	April 2019

Similar Records

Enhancement of polyhydroxyalkanoate production by co-feeding lignin derivatives with glycerol in Pseudomonas putida KT2440

Journal Article · Wed Jan 06 19:00:00 EST 2021 · Biotechnology for Biofuels · OSTI ID:1756127

Revealing oxidative pentose metabolism in new Pseudomonas putida isolates

Journal Article · Sat Dec 10 19:00:00 EST 2022 · Environmental Microbiology · OSTI ID:1903073

Related Subjects

09 BIOMASS FUELS

Engineered Pseudomonas putida KT2440 co-utilizes galactose and glucose

Citation Formats

References (34)

Similar Records

Related Subjects