Bioprocess development for muconic acid production from aromatic compounds and lignin

Salvachúa, Davinia; Johnson, Christopher W.; Singer, Christine A.; Rohrer, Holly; Peterson, Darren J.; Black, Brenna A.; Knapp, A; Beckham, Gregg T.

doi:10.1039/c8gc02519c

Cited by 132 publications

(102 citation statements)

References 51 publications

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“…It has been stated that industrially relevant production rates of fuels and chemicals, such as ethanol and muconic acid from biomass starting from a range of 1 g L −1 h −1 . When now considering a respective synthesis based on a secondary MET the microbial formate consumption rates are least in the range of 1 g L −1 h −1 (22 mmol L −1 h −1 ).…”

Section: Discussionsupporting

confidence: 77%

Engineering electrochemical CO₂ reduction to formate under bioprocess‐compatible conditions to bioreactor scale

2019

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The coupling of electrochemical CO 2 reduction to formate with the microbial biosynthesis of valuable products is promising for exploiting CO 2 as well as to store excess renewable electricity. Here, electrochemical CO 2 reduction to formate at bioprocesscompatible conditions was transferred from the scale of 50 mL electrochemical cells to 1 L electrobioreactors. In the 1 L electrobioreactor, a reproducible coulombic efficiency of 74.9 � 5.5 % and formate production rate of 0.038 � 0.005 mmol h À 1 cm À 2 could be achieved. Autoclaving by standard steam sterilization was shown to negatively affect the reliability of the process performance and, therefore, needs further investigation. However, mass transfer is not limiting for the process performance. Thus, the coupling of electrochemical CO 2 reduction to formate with microbial biosynthesis in sterile conditions at technical scale can be targeted in the future.[a] Dr.

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Section: Discussionsupporting

confidence: 77%

Engineering electrochemical CO₂ reduction to formate under bioprocess‐compatible conditions to bioreactor scale

2019

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“…This lignin comes from the solid fraction generated after enzymatic hydrolysis of pretreated corn stover (Chen et al ., ). Then, it is further washed with water (to remove sugars) and solubilized via base‐catalyzed depolymerization (Rodriguez et al ., ; Salvachúa et al ., ). Lignin solubilization was approximately 53% (lignin content in soluble stream/lignin content in initial solid stream) and contained ~ 4 g l − 1 of p ‐CA and 0.1‐0.2 g l − 1 ferulic acid (as major aromatic compounds) from an initial total lignin content of approximately 22 g l − 1 .…”

Section: Resultsmentioning

confidence: 97%

Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin

Salvachúa

Rydzak

Auwae

et al. 2019

Microbial Biotechnology

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SummaryMicrobial conversion offers a promising strategy for overcoming the intrinsic heterogeneity of the plant biopolymer, lignin. Soil microbes that natively harbour aromatic‐catabolic pathways are natural choices for chassis strains, and Pseudomonas putida KT2440 has emerged as a viable whole‐cell biocatalyst for funnelling lignin‐derived compounds to value‐added products, including its native carbon storage product, medium‐chain‐length polyhydroxyalkanoates (mcl‐PHA). In this work, a series of metabolic engineering targets to improve mcl‐PHA production are combined in the P. putida chromosome and evaluated in strains growing in a model aromatic compound, p‐coumaric acid, and in lignin streams. Specifically, the PHA depolymerase gene phaZ was knocked out, and the genes involved in β‐oxidation (fadBA1 and fadBA2) were deleted. Additionally, to increase carbon flux into mcl‐PHA biosynthesis, phaG, alkK, phaC1 and phaC2 were overexpressed. The best performing strain – which contains all the genetic modifications detailed above – demonstrated a 53% and 200% increase in mcl‐PHA titre (g l−1) and a 20% and 100% increase in yield (g mcl‐PHA per g cell dry weight) from p‐coumaric acid and lignin, respectively, compared with the wild type strain. Overall, these results present a promising strain to be employed in further process development for enhancing mcl‐PHA production from aromatic compounds and lignin.

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“…3b). Previous studies suggested that overexpression of vanAB promotes vanillic acid catabolism in P. putida KT2440 12 and A514 17 , respectively. Therefore, we inferred that increasing the expression of vanAB, which regenerates NAD + from NADH, should counter this hurdle (Fig.…”

Section: Resultsmentioning

confidence: 96%

“…To date, the biotransformation of ferulic acid to vanillin has been intensively studied, as vanillin is the aldehyde intermediate of the ferulic acid degradation pathway, and a widely used flavoring agent 8,10 . Recently there has been strong interest in the use of bacteria to produce more diverse and valuable products (e.g., polyhydroxyalkanoates (PHAs) and muconic acid) from ferulic acid 9,11,12 . PHAs, which act as microbial carbon storage, are a class of natural biopolymers and can be used as environmentally friendly alternatives to petroleum-based plastics, as well as bioimplant materials for medical and therapeutic applications [13][14][15][16] .…”

mentioning

confidence: 99%

Development of a CRISPR/Cas9n-based tool for metabolic engineering of Pseudomonas putida for ferulic acid-to-polyhydroxyalkanoate bioconversion

et al. 2020

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Ferulic acid is a ubiquitous phenolic compound in lignocellulose, which is recognized for its role in the microbial carbon catabolism and industrial value. However, its recalcitrance and toxicity poses a challenge for ferulic acid-to-bioproducts bioconversion. Here, we develop a genome editing strategy for Pseudomonas putida KT2440 using an integrated CRISPR/Cas9nλ-Red system with pyrF as a selection marker, which maintains cell viability and genetic stability, increases mutation efficiency, and simplifies genetic manipulation. Via this method, four functional modules, comprised of nine genes involved in ferulic acid catabolism and polyhydroxyalkanoate biosynthesis, were integrated into the genome, generating the KTc9n20 strain. After metabolic engineering and optimization of C/N ratio, polyhydroxyalkanoate production was increased to~270 mg/L, coupled with~20 mM ferulic acid consumption. This study not only establishes a simple and efficient genome editing strategy, but also offers an encouraging example of how to apply this method to improve microbial aromatic compound bioconversion.

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Bioprocess development for muconic acid production from aromatic compounds and lignin

Abstract: This work shows parallel strain and bioreactor process development to improve muconic acid production from aromatic compounds and lignin.

Cited by 132 publications

References 51 publications

Engineering electrochemical CO₂ reduction to formate under bioprocess‐compatible conditions to bioreactor scale

Engineering electrochemical CO₂ reduction to formate under bioprocess‐compatible conditions to bioreactor scale

Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin

Development of a CRISPR/Cas9n-based tool for metabolic engineering of Pseudomonas putida for ferulic acid-to-polyhydroxyalkanoate bioconversion

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Bioprocess development for muconic acid production from aromatic compounds and lignin

Abstract: This work shows parallel strain and bioreactor process development to improve muconic acid production from aromatic compounds and lignin.

Cited by 132 publications

References 51 publications

Engineering electrochemical CO2 reduction to formate under bioprocess‐compatible conditions to bioreactor scale

Engineering electrochemical CO2 reduction to formate under bioprocess‐compatible conditions to bioreactor scale

Metabolic engineering of Pseudomonas putida for increased polyhydroxyalkanoate production from lignin

Development of a CRISPR/Cas9n-based tool for metabolic engineering of Pseudomonas putida for ferulic acid-to-polyhydroxyalkanoate bioconversion

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Engineering electrochemical CO₂ reduction to formate under bioprocess‐compatible conditions to bioreactor scale

Engineering electrochemical CO₂ reduction to formate under bioprocess‐compatible conditions to bioreactor scale