Enhanced biosynthesis of d-tagatose from maltodextrin through modular pathway engineering of recombinant Escherichia coli

Dai, Yiwei; Li, Chenchen; Zheng, Luhua; Jiang, Bo; Zhang, Tao; Chen, Jingjing

doi:10.1016/j.bej.2021.108303

Cited by 14 publications

(11 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…rne131 encodes a missing RNase E protein that stabilizes many messenger ribonucleic acids, thus enhancing heterologous protein expression . Furthermore, the vectors pRSFDuet-1 and pETDuet-1 may be more compatible, and this combination also facilitates the synthesis of 2′-FL and d -tagatose; hence, combining pRSFDuet-1 and pETDuet-1 may be more suitable for the production of sugars, including LNT and LNnT. , In summary, the strains L0315 and LI0308, with a dual plasmid system of pRSFDuet-1 and pETDuet-1, were selected for the further construction of efficient strains for LNnT and LNT production.…”

Section: Resultsmentioning

confidence: 99%

“…coli and the production of the target product . The combinatorial optimization strategy of altering different plasmids has been widely used to produce HMOs and other target metabolites efficiently in vivo. − To achieve high LNnT and LNT production capabilities, lgtA and galE in pCDF-AE, HpgalT in pBS-HpgalT, and SewbdO in pBS–SewbdO were inserted into the pRSFDuet-1, pETDuet-1, and pACYCDuet-1 vectors, respectively, and 11 newly engineered strains for LNT and LNnT production were constructed according to different vector combinations (Figure A). These three vectors have copy numbers of >100, ∼40, and 10–12, respectively; in addition, pCDFDuet-1 and pBlueScript II SK (+) have copy numbers of 20–40 and 300–500, respectively. , …”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Metabolic Engineering of Escherichia coli for High-Level Production of Lacto-N-neotetraose and Lacto-N-tetraose

Liao

et al. 2023

J. Agric. Food Chem.

View full text Add to dashboard Cite

Lacto-N-neotetraose (LNnT) and lacto-N-tetraose (LNT) are important oligosaccharides found in breast milk and are commonly used as nutritional supplements in infant formula. We used metabolic engineering techniques to optimize the modified Escherichia coli BL21 star (DE3) strain for efficient synthesis of LNnT and LNT using β-1,4-galactosyltransferase (HpgalT) from Helicobacter pylori and β-1,3-galactosyltransferase (SewbdO) from Salmonella enterica subsp. salamae serovar, respectively. Further, we optimized the expression of three key genes, lgtA, galE, and HpgalT (SewbdO), to synthesize LNnT or LNT and deleted several genes (ugd, ushA, agp, wcaJ, otsA, and wcaC) to block competition in the UDP-galactose synthesis pathway. The optimized strain produced LNnT or LNT with a titer of 22.07 or 48.41 g/L, respectively, in a supplemented batch culture, producing 0.41 or 0.73 g/L/h, respectively. The strategies used in this study contribute to the development of cell factories for high-level LNnT and LNT and their derivatives.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Metabolic Engineering of Escherichia coli for High-Level Production of Lacto-N-neotetraose and Lacto-N-tetraose

Liao

et al. 2023

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

“…To further demonstrate that the novel foam materials can be used for biocatalytic production processes, we chose to produce a challenging product, the rare sugar tagatose. [37] Commonly L-arabinose isomerase is being used as either free or immobilized enzyme, or in whole-cell catalysis, to produce D-tagatose by isomerization from D-galactose, leading to productivities for free or immobilized isomerase of 0.3 -9.6 g L −1 h −1 [38][39][40][41][42] and of 0.21 -1.13 g L −1 h −1 for synthesis by whole-cells. [37,43,44] Recently, oxidoreductive reactions have been used to overcome the thermodynamic equilibrium of the isomerization reaction and to improve the purification of the final product.…”

Section: Toward Applicationsmentioning

confidence: 99%

Biocatalytic Foams from Microdroplet‐Formulated Self‐Assembling Enzymes

Hertel,

Bitterwolf,

Kröll

et al. 2023

Advanced Materials

View full text Add to dashboard Cite

Industrial biocatalysis plays an important role in the development of a sustainable economy, as enzymes can be used to synthesize an enormous range of complex molecules under environmentally friendly conditions. To further develop the field, intensive research is being conducted on process technologies for continuous flow biocatalysis in order to immobilize large quantities of enzyme biocatalysts in microstructured flow reactors under conditions that are as gentle as possible in order to realize efficient material conversions. Here, monodisperse foams consisting almost entirely of enzymes covalently linked via SpyCatcher/SpyTag conjugation are reported. The biocatalytic foams are readily available from recombinant enzymes via microfluidic air‐in‐water droplet formation, can be directly integrated into microreactors, and can be used for biocatalytic conversions after drying. Reactors prepared by this method show surprisingly high stability and biocatalytic activity. The physicochemical characterization of the new materials is described and exemplary applications in biocatalysis are shown using two‐enzyme cascades for the stereoselective synthesis of chiral alcohols and the rare sugar tagatose.

show abstract

“…PMM/PGMs have received interest over the years in the context of their use in enzyme-based production of biochemicals with different potential industrial applications (Wei et al, 2021;Dai et al, 2022). For example, a heat stable PGM which has been characterized in Clostridium thermocellum (Wang and Zhang, 2010) has recently been used to convert glucose 1 phosphate to glucose 6-phosphate in an enzymatic cascade involving five other enzymes in a one-pot stoichiometric conversion of starch to mannitol (Wei et al, 2021).…”

Section: Biotechnological Potentialmentioning

confidence: 99%

“…This system has the potential to be further expanded to use starch for the efficient synthesis of other valueadded biochemicals (Wei et al, 2021). Similarly, the PGM from Thermococcus kodakaraensis has also recently been used in a recombinant E. coli vector system for the production of another food sweetener, namely D-tagatose from maltodextrin using whole-cell catalysts (Dai et al, 2022); while a thermostable PGM from T. kodakaraensis has been used in the production of myo-inositol, an important human dietary supplement, from starch (Fujisawa et al, 2017). A recombinant PGM from E. coli has also been used to enhance UDP-galactose derived synthesis of the bioactive compound hyperoside (quercetin 3-O-galactoside) in a metabolically engineered E. coli strain (Li et al, 2022).…”

Section: Biotechnological Potentialmentioning

confidence: 99%

Isolation, identification, and biochemical characterization of a novel bifunctional phosphomannomutase/phosphoglucomutase from the metagenome of the brown alga Laminaria digitata

et al. 2022

View full text Add to dashboard Cite

Macroalgae host diverse epiphytic bacterial communities with potential symbiotic roles including important roles influencing morphogenesis and growth of the host, nutrient exchange, and protection of the host from pathogens. Macroalgal cell wall structures, exudates, and intra-cellular environments possess numerous complex and valuable carbohydrates such as cellulose, hemi-cellulose, mannans, alginates, fucoidans, and laminarin. Bacterial colonizers of macroalgae are important carbon cyclers, acquiring nutrition from living macroalgae and also from decaying macroalgae. Seaweed epiphytic communities are a rich source of diverse carbohydrate-active enzymes which may have useful applications in industrial bioprocessing. With this in mind, we constructed a large insert fosmid clone library from the metagenome of Laminaria digitata (Ochrophyta) in which decay was induced. Subsequent sequencing of a fosmid clone insert revealed the presence of a gene encoding a bifunctional phosphomannomutase/phosphoglucomutase (PMM/PGM) enzyme 10L6AlgC, closely related to a protein from the halophilic marine bacterium, Cobetia sp. 10L6AlgC was subsequently heterologously expressed in Escherichia coli and biochemically characterized. The enzyme was found to possess both PMM and PGM activity, which had temperature and pH optima of 45°C and 8.0, respectively; for both activities. The PMM activity had a Km of 2.229 mM and Vmax of 29.35 mM min−1 mg−1, while the PGM activity had a Km of 0.5314 mM and a Vmax of 644.7 mM min−1 mg−1. Overall characterization of the enzyme including the above parameters as well as the influence of various divalent cations on these activities revealed that 10L6AlgC has a unique biochemical profile when compared to previously characterized PMM/PGM bifunctional enzymes. Thus 10L6AlgC may find utility in enzyme-based production of biochemicals with different potential industrial applications, in which other bacterial PMM/PGMs have previously been used such as in the production of low-calorie sweeteners in the food industry.

show abstract

Enhanced biosynthesis of d-tagatose from maltodextrin through modular pathway engineering of recombinant Escherichia coli

Cited by 14 publications

References 23 publications

Metabolic Engineering of Escherichia coli for High-Level Production of Lacto-N-neotetraose and Lacto-N-tetraose

Metabolic Engineering of Escherichia coli for High-Level Production of Lacto-N-neotetraose and Lacto-N-tetraose

Biocatalytic Foams from Microdroplet‐Formulated Self‐Assembling Enzymes

Isolation, identification, and biochemical characterization of a novel bifunctional phosphomannomutase/phosphoglucomutase from the metagenome of the brown alga Laminaria digitata

Contact Info

Product

Resources

About