Glyoxylate carboligase-based whole-cell biotransformation of formaldehyde into ethylene glycol <i>via</i> glycolaldehyde

Jo, Hye-Jin; Kim, Junhong; Kim, Ye-Na; Seo, Pil-Won; Kim, Chae‐Yun; Kim, Jiwon; Yu, Han-na; Cheon, Huijin; Lee, Eun Yeol; Kim, Ji Soo; Park, Jin Byung

doi:10.1039/d1gc03549e

Cited by 26 publications

(33 citation statements)

References 46 publications

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“…However, no proof of in vivo functionality of this pathway was shown. More recently a new GalS was engineered based on a newly discovered glyoxylate carboligase enzyme found in E. coli ( Jo et al, 2022 ). One of the variants showed a 10 times higher affinity and a 2 times higher catalytic efficiency for formaldehyde compared to the previously described GalS ( Lu, et al, 2019 ; Yang, et al, 2019 ).…”

Section: Create Novel Enzymesmentioning

confidence: 99%

“…One of the variants showed a 10 times higher affinity and a 2 times higher catalytic efficiency for formaldehyde compared to the previously described GalS ( Lu, et al, 2019 ; Yang, et al, 2019 ). When this optimized GalS was associated in vitro to a lactaldehyde reductase (FucO) , 66% of the formaldehyde was converted into ethylene glycol ( Jo, et al, 2022 ).…”

Section: Create Novel Enzymesmentioning

confidence: 99%

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From a Hetero- to a Methylotrophic Lifestyle: Flash Back on the Engineering Strategies to Create Synthetic Methanol-User Strains

Peiro

Vicente

Jallet

et al. 2022

Front. Bioeng. Biotechnol.

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Engineering microorganisms to grow on alternative feedstocks is crucial not just because of the indisputable biotechnological applications but also to deepen our understanding of microbial metabolism. One-carbon (C1) substrate metabolism has been the focus of extensive research for the prominent role of C1 compounds in establishing a circular bioeconomy. Methanol in particular holds great promise as it can be produced directly from greenhouse gases methane and carbon dioxide using renewable resources. Synthetic methylotrophy, i.e. introducing a non-native methanol utilization pathway into a model host, has therefore been the focus of long-time efforts and is perhaps the pinnacle of metabolic engineering. It entails completely changing a microorganism’s lifestyle, from breaking up multi-carbon nutrients for growth to building C-C bonds from a single-carbon molecule to obtain all metabolites necessary to biomass formation as well as energy. The frontiers of synthetic methylotrophy have been pushed further than ever before and in this review, we outline the advances that paved the way for the more recent accomplishments. These include optimizing the host’s metabolism, “copy and pasting” naturally existing methylotrophic pathways, “mixing and matching” enzymes to build new pathways, and even creating novel enzymatic functions to obtain strains that are able to grow solely on methanol. Finally, new approaches are contemplated to further advance the field and succeed in obtaining a strain that efficiently grows on methanol and allows C1-based production of added-value compounds.

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Section: Create Novel Enzymesmentioning

confidence: 99%

Section: Create Novel Enzymesmentioning

confidence: 99%

From a Hetero- to a Methylotrophic Lifestyle: Flash Back on the Engineering Strategies to Create Synthetic Methanol-User Strains

Peiro

Vicente

Jallet

et al. 2022

Front. Bioeng. Biotechnol.

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“…The gene coding for EcGCL from E. coli K-12 and variants were constructed as previously 15 . The AldA from E. coli K-12 was ampli ed by PCR from chromosomal DNA of E. coli K-12 using the primers which were designed for In-Fusion cloning protocol (Takara Bio.…”

Section: Gene Cloning and Construction Of Recombinant Plasmidsmentioning

confidence: 99%

“…Two molecules of formaldehyde are condensed into one molecule of glycolaldehyde by a carboligase, which is further transformed into glycolic acid by an aldehyde dehydrogenase (Scheme 1). A number of carboligases including formolase 3,21 and glyoxylate carboligase were examined for the condensation of two molecules of formaldehyde into glycolaldehyde 15 . A formolase variant (i.e., FLSH29DQ113C) 21 was better than parent enzyme in terms of the production yield of glycolaldehyde from formaldehyde.…”

Section: Biotransformation Of Formaldehyde Into Glycolic Acidmentioning

confidence: 99%

“…The concentrations of reaction substrates and products were determined as previously. 15 Brie y, The formaldehyde concentrations were assayed by a modi cation of the Nash reaction for 96-well plate 19 . A 100 µL of diluted sample and 100 µL of Nash reagent (0.2 % 2,4-pentanedione, 0.1 M acetic acid, and 3.89 M ammonium acetate) was incubated at 37 °C for 30 min.…”

Section: Reactant Analysismentioning

confidence: 99%

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Biotransformation pathway and side reaction engineering for the whole-cell production of glycolic acid from formaldehyde

Kim

et al. 2022

Preprint

Self Cite

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A very simple biocatalytic system for the synthesis of the industrially relevant C2 chemicals (e.g., glycolic acid (3)) from formaldehyde (1) was established by whole-cell biocatalyst. The biocatalytic system consisted of an engineered glyoxylate carboligase from Escherichia coli K-12 (EcGCL) and an aldehyde dehydrogenase (e.g., AldA from E. coli K-12). To improve glycolic acid production, we have investigated metabolic engineering to regulate substrate consumption and product bypass and E. coli-based strain optimization. The target compound was produced at rates of up to 20.3 U/g dry cells with conversions up to 92%. This study will ensure the sustainability of the green energy industry by synthesizing valuable C2 chemical from a single carbon compound, formaldehyde.

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Recent Progress in the Synthesis of α‐Hydroxy Carbonyl Compounds with ThDP‐dependent Carboligases

Dobiašová,

Jurkaš,

Both

et al. 2024

ChemCatChem

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Carboligases catalyze the thiamine diphosphate (ThDP) dependent formation of carbon‐carbon bonds. These enzymes are prominent biocatalysts for the production of valuable a‐hydroxy carbonyl compounds, which serve as key building blocks in the synthesis of various pharmaceuticals and fine chemicals. Carboligases act in selective manner to afford regio‐ and stereochemically defined products from simple starting materials. This review explores the catalytic prowess of carboligases for synthetic purposes and is aimed at providing a selection tool of enzymes for particular sets of substrates or desired products. The comprehensive overview encompasses structural insights, relationships of the currently known sequence space and practical applications with a focus on recent literature, showcasing carboligases as potent tools for sustainable and efficient synthesis.

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Glyoxylate carboligase-based whole-cell biotransformation of formaldehyde into ethylene glycol via glycolaldehyde

Abstract: A novel biocatalytic system for the synthesis of the industrially relevant C2 chemicals (e.g., ethylene glycol (3)) from formaldehyde (1) was established. The biocatalytic system consisted of a newly discovered...

Cited by 26 publications

References 46 publications

From a Hetero- to a Methylotrophic Lifestyle: Flash Back on the Engineering Strategies to Create Synthetic Methanol-User Strains

From a Hetero- to a Methylotrophic Lifestyle: Flash Back on the Engineering Strategies to Create Synthetic Methanol-User Strains

Biotransformation pathway and side reaction engineering for the whole-cell production of glycolic acid from formaldehyde

Recent Progress in the Synthesis of α‐Hydroxy Carbonyl Compounds with ThDP‐dependent Carboligases

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