Engineered Pseudomonas putida simultaneously catabolizes five major components of corn stover lignocellulose: Glucose, xylose, arabinose, p-coumaric acid, and acetic acid

Elmore, Joshua R.; Dexter, Gara N.; Salvachúa, Davinia; O’Brien, Marykate H.; Klingeman, Dawn M.; Gorday, Kent; Michener, Joshua K.; Peterson, Darren J.; Beckham, Gregg T.; Guss, Adam M.

doi:10.1016/j.ymben.2020.08.001

Cited by 76 publications

(105 citation statements)

References 48 publications

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“…In a related study, the Crc protein was identified as a global regulator whose inactivation has been demonstrated to improve product formation (cis, cis muconic acid) from p CA under laboratory cultivation conditions 13 . However, when translated to an aromatics and sugar co-utilizing engineered P. putida strain, crc deficient cells exhibited a significant lag phase under bioreactor growth conditions 41 . As we did not identify crc as a positive fitness mutant in our analysis of bioreactor-advantaged strains, we instead argue optimizing strains for bioproduct formation using laboratory settings may be inadequate; the same optimizations can have negative implications upon scale-up.…”

Section: Discussionmentioning

confidence: 99%

“…The ambr ® 250 improves our throughput to up to 12 simultaneous stirred tank runs but increasing the throughput above 24 units requires a significant capital investment. The isogenic deletion strain collection of bioreactor-advantaged mutants is ready to be screened with other heterologous gene pathways and carbon streams, such as xylose 41 . This functional genomics data can also help improve predictability of machine learning tools, such as ART 53 .…”

Section: Discussionmentioning

confidence: 99%

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High Throughput Fitness Profiling Reveals Loss Of GacS-GacA Regulation Improves Indigoidine Production InPseudomonas putida

Eng

Banerjee

Lau

et al. 2021

Preprint

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Pseudomonas putida KT2440 is an emerging industrial microbe amenable for use with renewable carbon streams including aromatics such as para-coumarate (pCA). We examined this microbe under common stirred-tank bioreactor parameters with quantitative fitness assays using a pooled transposon library containing nearly all (4,778) non-essential genes. Assessing differential fitness values by monitoring changes in mutant strain abundance over time identified 31 genes with improved fitness in multiple bioreactor-relevant parameters. Twenty-one genes from this subset were reconstructed, including GacA, a signaling protein, TtgB, an ABC transporter, and PP_0063, a lipid A acyltransferase. Twelve deletion strains with roles in varying cellular functions were evaluated for conversion of pCA, to a heterologous bioproduct, indigoidine. Several mutants, such as the ΔgacA strain improved both fitness in a bioreactor and showed an 8-fold improvement in indigoidine production (4.5 g/L, 0.29 g/g pCA, 23% MTY) from pCA as the carbon source.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

High Throughput Fitness Profiling Reveals Loss Of GacS-GacA Regulation Improves Indigoidine Production InPseudomonas putida

Eng

Banerjee

Lau

et al. 2021

Preprint

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“…CBP has focused on fermentative microbes that can deconstruct the carbohydrate fraction of lignocellulose, with less focus on lignin deconstruction due to the associated difficulties. However, some organisms are being studied for their ability to process lignin derivatives, such as Pseudomonas putida (Elmore et al ., 2020).…”

Section: Cellulolytic and Hemicellulolytic Microorganisms With Industrial Potentialmentioning

confidence: 99%

Thermophilic microbial deconstruction and conversion of natural and transgenic lignocellulose

Bing

Sulis

Wang

et al. 2021

Environ Microbiol Rep

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The potential to convert renewable plant biomasses into fuels and chemicals by microbial processes presents an attractive, less environmentally intense alternative to conventional routes based on fossil fuels. This would best be done with microbes that natively deconstruct lignocellulose and concomitantly form industrially relevant products, but these two physiological and metabolic features are rarely and simultaneously observed in nature. Genetic modification of both plant feedstocks and microbes can be used to increase lignocellulose deconstruction capability and generate industrially relevant products. Separate efforts on plants and microbes are ongoing, but these studies lack a focus on optimal, complementary combinations of these disparate biological systems to obtain a convergent technology. Improving genetic tools for plants have given rise to the generation of low-lignin lines that are more readily solubilized by microorganisms. Most focus on the microbiological front has involved thermophilic bacteria from the genera Caldicellulosiruptor and Clostridium, given their capacity to degrade lignocellulose and to form bio-products through metabolic engineering strategies enabled by ever-improving molecular genetics tools. Bioengineering plant properties to better fit the deconstruction capabilities of candidate consolidated bioprocessing microorganisms has potential to achieve the efficient lignocellulose deconstruction needed for industrial relevance.

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“…Xylose, a pentose, is the major constituent of lignocellulose biomass, and was deemed to be the second most abundant biological resource, after glucose (Elmore et al, 2020). High-value utilization of xylose is shifting from bioenergy to a broader range of chemicals.…”

Section: Bioconversion Of Single-carbon Sources To Valuable Chemicalsmentioning

confidence: 99%

Recent Progress on Chemical Production From Non-food Renewable Feedstocks Using Corynebacterium glutamicum

Zhang

Jiang

et al. 2020

Front. Bioeng. Biotechnol.

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Due to the non-renewable nature of fossil fuels, microbial fermentation is considered a sustainable approach for chemical production using glucose, xylose, menthol, and other complex carbon sources represented by lignocellulosic biomass. Among these, xylose, methanol, arabinose, glycerol, and other alternative feedstocks have been identified as superior non-food sustainable carbon substrates that can be effectively developed for microbe-based bioproduction. Corynebacterium glutamicum is a model gram-positive bacterium that has been extensively engineered to produce amino acids and other chemicals. Recently, in order to reduce production costs and avoid competition for human food, C. glutamicum has also been engineered to broaden its substrate spectrum. Strengthening endogenous metabolic pathways or assembling heterologous ones enables C. glutamicum to rapidly catabolize a multitude of carbon sources. This review summarizes recent progress in metabolic engineering of C. glutamicum toward a broad substrate spectrum and diverse chemical production. In particularly, utilization of lignocellulosic biomass-derived complex hybrid carbon source represents the futural direction for non-food renewable feedstocks was discussed.

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Engineered Pseudomonas putida simultaneously catabolizes five major components of corn stover lignocellulose: Glucose, xylose, arabinose, p-coumaric acid, and acetic acid

Cited by 76 publications

References 48 publications

High Throughput Fitness Profiling Reveals Loss Of GacS-GacA Regulation Improves Indigoidine Production InPseudomonas putida

High Throughput Fitness Profiling Reveals Loss Of GacS-GacA Regulation Improves Indigoidine Production InPseudomonas putida

Thermophilic microbial deconstruction and conversion of natural and transgenic lignocellulose

Recent Progress on Chemical Production From Non-food Renewable Feedstocks Using Corynebacterium glutamicum

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