Combinatorial synthetic pathway fine-tuning and cofactor regeneration for metabolic engineering of Escherichia coli significantly improve production of D-glucaric acid

Su, Huihui; Peng, Fei; Ou, Xiao‐Yang; Zeng, Ying‐Jie; Zong, Min‐Hua; Lou, Wen‐Yong

doi:10.1016/j.nbt.2020.03.004

Cited by 11 publications

(14 citation statements)

References 33 publications

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“…The alternative is bio-based production approaches. Therefore, the development of GA production based on fermentation and metabolically engineered cells or in vitro cell-free biocatalysis has leapt forward in recent years, and GA titers and yield have increased rapidly [ 18 , 19 , 26 , 40 , 51 , 64 , 69 ]. Firstly, a series of synthetic pathways for GA have been ported into E. coli , S. cerevisiae , and P. pastoris , and these engineered cells show the ability to synthesize GA de novo.…”

Section: Discussionmentioning

confidence: 99%

“…Ribosome binding site (RBS) optimization is another strategy to enhance GA production by coordinating the expression of enzymes [ 39 ]. The activity ratio of Ino1 to MIOX was improved from 1:0.40 to 1:3.41, which greatly enhanced the GA titer starting from glucose, to 4.56 g/L, compared with the unoptimized strain (3.42 g/L) and the yield of GA was 0.467 g/g glucose [ 40 ].…”

Section: Construction and Optimization Of Heterologous Ga Synthesis Pmentioning

confidence: 99%

“…The increase of cellular oxygen might increase oxygen availability for this reaction and/or accelerate cell metabolism and/or growth and thus cause more efficient GA production [ 58 ]. It is worth mentioning that when P. pastoris was employed as the host, recombinant strains produced GA from glucose and MI up to 6.61 ± 0.30 g/L, which is also better than the titer achieved in recombinant E. coli [ 40 ].…”

Section: Construction and Optimization Of Heterologous Ga Synthesis Pmentioning

confidence: 99%

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Cell-based and cell-free biocatalysis for the production of d-glucaric acid

Chen

Huang²,

Hou

et al. 2020

Biotechnol Biofuels

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Abstractd-Glucaric acid (GA) is a value-added chemical produced from biomass, and has potential applications as a versatile platform chemical, food additive, metal sequestering agent, and therapeutic agent. Marketed GA is currently produced chemically, but increasing demand is driving the search for eco-friendlier and more efficient production approaches. Cell-based production of GA represents an alternative strategy for GA production. A series of synthetic pathways for GA have been ported into Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris, respectively, and these engineered cells show the ability to synthesize GA de novo. Optimization of the GA metabolic pathways in host cells has leapt forward, and the titer and yield have increased rapidly. Meanwhile, cell-free multi-enzyme catalysis, in which the desired pathway is constructed in vitro from enzymes and cofactors involved in GA biosynthesis, has also realized efficient GA bioconversion. This review presents an overview of studies of the development of cell-based GA production, followed by a brief discussion of potential applications of biosensors that respond to GA in these biosynthesis routes.

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Section: Discussionmentioning

confidence: 99%

Section: Construction and Optimization Of Heterologous Ga Synthesis Pmentioning

confidence: 99%

Section: Construction and Optimization Of Heterologous Ga Synthesis Pmentioning

confidence: 99%

See 1 more Smart Citation

Cell-based and cell-free biocatalysis for the production of d-glucaric acid

Chen

Huang²,

Hou

et al. 2020

Biotechnol Biofuels

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“…S. cerevisiae can be used for production of glucarate by coexpression of four key enzymes including myo -inositol-1-phosphate synthase (MIPS), inositol-1-monophosphatase (IMPase), myo -inositol oxygenase (MIOX), and uronic acid/uronate dehydrogenase (UDH) to produce glucaric acid from extracellular glucose (Figure ). By fed-batch fermentation in a 5-L bioreactor, the bioconversion of glucose to glucaric acid could be achieved with a yield of 0.20 g/g and production rate at 0.03 g/L h. Recently, a biosynthetic pathway for glucaric acid production in E. coli from raw glucose was established by overexpression of the same four key enzymes as in S. cerevisiae . The flux of the glucaric acid biosynthetic pathway could be enhanced by blocking the conversion of glucose into G6P by disrupting G6PD and GPI expression.…”

Section: Production Of Lignocellulosic Biomass-derived Productsmentioning

confidence: 99%

“…229 By fed-batch fermentation in a 5-L bioreactor, the bioconversion of glucose to glucaric acid could be achieved with a yield of 0.20 g/g and production rate at 0.03 g/L h. Recently, a biosynthetic pathway for glucaric acid production in E. coli from raw glucose was established by overexpression of the same four key enzymes as in S. cerevisiae. 230 The flux of the glucaric acid biosynthetic pathway could be enhanced by blocking the conversion of glucose into G6P by disrupting G6PD and GPI expression. Moreover, genes expressing uronic acid isomerase (UI) and glucaric acid/ glucarate dehydratase (GADHT) were deleted to increase the glucarate production titer.…”

Section: Production Of Lignocellulosic Biomass-derived Productsmentioning

confidence: 99%

Enzymes, In Vivo Biocatalysis, and Metabolic Engineering for Enabling a Circular Economy and Sustainability

et al. 2021

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Since the industrial revolution, the rapid growth and development of global industries have depended largely upon the utilization of coal-derived chemicals, and more recently, the utilization of petroleum-based chemicals. These developments have followed a linear economy model (produce, consume, and dispose). As the world is facing a serious threat from the climate change crisis, a more sustainable solution for manufacturing, i.e., circular economy in which waste from the same or different industries can be used as feedstocks or resources for production offers an attractive industrial/business model. In nature, biological systems, i.e., microorganisms routinely use their enzymes and metabolic pathways to convert organic and inorganic wastes to synthesize biochemicals and energy required for their growth. Therefore, an understanding of how selected enzymes convert biobased feedstocks into special (bio)chemicals serves as an important basis from which to build on for applications in biocatalysis, metabolic engineering, and synthetic biology to enable biobased processes that are greener and cleaner for the environment. This review article highlights the current state of knowledge regarding the enzymatic reactions used in converting biobased wastes (lignocellulosic biomass, sugar, phenolic acid, triglyceride, fatty acid, and glycerol) and greenhouse gases (CO 2 and CH 4 ) into value-added products and discusses the current progress made in their metabolic engineering. The commercial aspects and life cycle assessment of products from enzymatic and metabolic engineering are also discussed. Continued development in the field of metabolic engineering would offer diversified solutions which are sustainable and renewable for manufacturing valuable chemicals.

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Production of d-glucaric acid with phosphoglucose isomerase-deficient Saccharomyces cerevisiae

Toivari,

Vehkomäki,

Ruohonen

et al. 2023

Biotechnol Lett

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Abstractd-Glucaric acid is a potential biobased platform chemical. Previously mainly Escherichia coli, but also the yeast Saccharomyces cerevisiae, and Pichia pastoris, have been engineered for conversion of d-glucose to d-glucaric acid via myo-inositol. One reason for low yields from the yeast strains is the strong flux towards glycolysis. Thus, to decrease the flux of d-glucose to biomass, and to increase d-glucaric acid yield, the four step d-glucaric acid pathway was introduced into a phosphoglucose isomerase deficient (Pgi1p-deficient) Saccharomyces cerevisiae strain. High d-glucose concentrations are toxic to the Pgi1p-deficient strains, so various feeding strategies and use of polymeric substrates were studied. Uniformly labelled 13C-glucose confirmed conversion of d-glucose to d-glucaric acid. In batch bioreactor cultures with pulsed d-fructose and ethanol provision 1.3 g d-glucaric acid L−1 was produced. The d-glucaric acid titer (0.71 g d-glucaric acid L−1) was lower in nitrogen limited conditions, but the yield, 0.23 g d-glucaric acid [g d-glucose consumed]−1, was among the highest that has so far been reported from yeast. Accumulation of myo-inositol indicated that myo-inositol oxygenase activity was limiting, and that there would be potential to even higher yield. The Pgi1p-deficiency in S. cerevisiae provides an approach that in combination with other reported modifications and bioprocess strategies would promote the development of high yield d-glucaric acid yeast strains.

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Combinatorial synthetic pathway fine-tuning and cofactor regeneration for metabolic engineering of Escherichia coli significantly improve production of D-glucaric acid

Cited by 11 publications

References 33 publications

Cell-based and cell-free biocatalysis for the production of d-glucaric acid

Cell-based and cell-free biocatalysis for the production of d-glucaric acid

Enzymes, In Vivo Biocatalysis, and Metabolic Engineering for Enabling a Circular Economy and Sustainability

Production of d-glucaric acid with phosphoglucose isomerase-deficient Saccharomyces cerevisiae

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