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L-Cysteine and L-methionine, as the only two sulfur-containing amino acids among the canonical 20 amino acids, possess distinct characteristics and find wide-ranging industrial applications. The use of different organisms for fermentative production of L-cysteine and L-methionine is gaining increasing attention, with Escherichia coli being extensively studied as the preferred strain. This preference is due to its ability to grow rapidly in cost-effective media, its robustness for industrial processes, the well-characterized metabolism, and the availability of molecular tools for genetic engineering. This review focuses on the genetic and molecular mechanisms involved in the production of these sulfur-containing amino acids in E. coli. Additionally, we systematically summarize the metabolic engineering strategies employed to enhance their production, including the identification of new targets, modulation of metabolic fluxes, modification of transport systems, dynamic regulation strategies, and optimization of fermentation conditions. The strategies and design principles discussed in this review hold the potential to facilitate the development of strain and process engineering for direct fermentation of sulfur-containing amino acids., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2023. Published by Elsevier Inc.)

A new composite sponge assisted by magnetic field-mediated guidance was developed for effective hemostasis. It was based on polydopamine capillary-channel agarose (PDA-CAGA) sponge as matrix; meanwhile, the combination of deep eutectic solvent (DES, choline chloride:glycerol = 1:1, M/M)-dispersed Fe 3 O 4 nanoparticles after fabrication by tannic acid (DES-Fe 3 O 4 @TA) was applied as hemostatic magnetic fluid. This sponge had oriented and aligned capillary channels realized by a 3D printed pattern, which endowed them with obvious shape memory and liquid absorption performance. Computational simulation was performed to describe the fluid status in channels; DES-Fe 3 O 4 @TA exhibited good magnetic properties, fluidity, and stability. In addition, the sponge driven to react rapidly with the bleeding site under the effect of a magnetic field presented a shorter hemostasis time (reduced by 85.02% in the tail and 81.07% in the liver of rats) and less blood loss (reduced by 97.08% in the tail and 91.50% in the liver) than those of medical gelatin sponge (GS). Meanwhile, the multifunctional material also exhibited better biocompatibility, procoagulant performance, and significant inhibition on S. aureus and E. coli than GS. As a whole, this work proposed a new strategy for rapid hemostasis by designing a magnetic field assisted composite bacteriostatic material, which also expanded the applications of green solvents in the clinical management field.

The rigidity of bacterial cell walls synthesized by a complicated pathway limit the cell shapes as coccus, bar or ellipse or even fibers. A less rigid bacterium could be beneficial for intracellular accumulation of poly-3-hydroxybutyrate (PHB) as granular inclusion bodies. To understand how cell rigidity affects PHB accumulation, E. coli cell wall synthesis pathway was reinforced and weakened, respectively. Cell rigidity was achieved by thickening the cell walls via insertion of a constitutive gltA (encoding citrate synthase) promoter in front of a series of cell wall synthesis genes on the chromosome of several E. coli derivatives, resulting in 1.32-1.60 folds increase of Young's modulus in mechanical strength for longer E. coli cells over-expressing fission ring FtsZ protein inhibiting gene sulA. Cell rigidity was weakened by down regulating expressions of ten genes in the cell wall synthesis pathway using CRISPRi, leading to elastic cells with more spaces for PHB accumulation. The regulation on cell wall synthesis changes the cell rigidity: E. coli with thickened cell walls accumulated only 25% PHB while cell wall weakened E. coli produced 93% PHB. Manipulation on cell wall synthesis mechanism adds another possibility to morphology engineering of microorganisms., (Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)

Maltose-binding protein (MBP) is a component of the maltose transport system of Escherichia coli. Our previous study found that MBP combined with Bacillus Calmette-Guerin (BCG) increases the percentage of activated macrophages in the spleen and the pinocytic activity of peritoneal macrophages in vivo. However, the effect of MBP alone on macrophages remains unclear. In the present study, the results showed that MBP enhanced LPS-stimulated macrophage activity in vivo. Subsequently, we investigated the regulatory effect of MBP on mouse peritoneal macrophages in vitro and the possible underlying mechanism. The results showed that MBP directly promoted macrophage phagocytic activity and increased the production of NO, IL-1β and IL-6. Notably, macrophage phenotypic analysis showed that MBP significantly increased iNOS, IL-12p70 and CD16/32. In contrast, MBP decreased the secretion of IL-10 and slightly decreased Arg-1 mRNA and CD206 protein expression. These results suggested that MBP activated macrophages and polarized them into M1 macrophages. Further study found that MBP directly bound to macrophages and upregulated TLR2 mRNA expression. This process was accompanied by a clear increase in MyD88 expression and phosphorylation of p38 MAPK and IκB-α, but these effects were largely abrogated by pretreatment with anti-TLR2 or anti-TLR4 antibodies. The effects of MBP on macrophage NO production were also partially inhibited by anti-TLR2 and/or anti-TLR4 antibodies. Furthermore, the effect of MBP on IL-12 and IL-10 secretion was largely influenced by the NF-κB inhibitor PDTC and the p38 MAPK inhibitor SB203580. These results suggest that MBP directly activates macrophages and induces M1 polarization through a process that may involve TLR2 and TLR4., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Bacteria, exemplified by enteropathogenic Escherichia coli (E. coli), rely on elaborate acid resistance systems to survive acidic environment (such as the stomach). Comprehensive understanding of bacterial acid resistance is important for prevention and clinical treatment. In this study, we report a previously uncharacterized type of acid resistance system in E. coli that relies on L-glutamine (Gln), one of the most abundant food-borne free amino acids. Upon uptake into E. coli, Gln is converted to L-glutamate (Glu) by the acid-activated glutaminase YbaS, with concomitant release of gaseous ammonia. The free ammonia neutralizes proton, resulting in elevated intracellular pH under acidic environment. We show that YbaS and the amino acid antiporter GadC, which exchanges extracellular Gln with intracellular Glu, together constitute an acid resistance system that is sufficient for E. coli survival under extremely acidic environment.

• A recombinant strain E. coli DH5α J23106 overexpressing merB gene was constructed to degrade MeHg. • DH5α J23106 degraded MeHg more than 81.6% in culture medium under anoxic/oxic conditions. • DH5α J23106 degrade newly produced and intrinsic MeHg in actual Hg-contaminated sediment. • Anoxic/oxic aging of MeHg in sediment decreased its degradation by DH5α J23106. • Ammonium thiosulfate leaching, combined with DH5α J23106, enhanced the degradation of MeHg in sediment. Sediment is the primary hotspot for microbial production of toxic and bio-accumulative methylmercury (MeHg). Common remediation strategies such as sediment dredging and capping can be too expensive and cannot degrade MeHg efficiently. Here, we constructed an Escherichia coli strain overexpressing merB gene (DH5α J23106) and assessed the effectiveness of this recombinant strain in degradation of MeHg in culture medium and sediment. DH5α J23106 can efficiently degrade MeHg (with initial concentration from 0.01 to 50 ng/mL) to more than 81.6% in a culture medium under anoxic and oxic conditions. Enriched isotope addition (199HgCl 2) revealed that this recombinant strain can degrade 78.6% of newly produced Me199Hg in actual sediment, however the biodegradation decreased to 36.3% for intrinsic MeHg. Degradation of spiked MeHg after aging in anoxic and oxic sediments further demonstrated DH5α J23106 can efficiently degrade newly produced MeHg and the degradation decreased with aging significantly, especially for oxic sediment. Eight sediments were further assessed for the biodegradation of aged MeHg by DH5α J23106 under oxic conditions, with degradation ratios ranging from 9.0% to 66.9%. When combined with (NH 4) 2 S 2 O 3 leaching, the degradation of MeHg increased by 15.8–38.8% in on-site and off-site modes through enhanced MeHg bioavailability in some of these sediments. Thus, this recombinant strain DH5α J23106 can degrade MeHg efficiently and have the potential for remediating bioavailable MeHg in contaminated sediments. [Display omitted] [ABSTRACT FROM AUTHOR]

A novel portable conductive system of hydroxylated fullerene (TEGs-C 60) embedded on the deep eutectic solvent (DES, choline chloride/malic acid)-agarose gel was developed for on-demand curation with skin contact. It has been found with better elasticity, conductivity and thermal stability than agarose hydrogel. Especially, the electro-responsive drug releasing amount on the skin of mice and live rabbits through small batteries was proved to be 4–5 times of those without electrical stimulation, which reached the level of clinic daily dosage. Meanwhile, it also showed good biocompatibility and much higher antibacterial rate against S. aureus (78.67 ± 0.80 %) together with E. coli (92.08 ± 1.50 %) than those of commercial chitosan gel. The new electrical "switch" gel system provided an effective and useful development strategy for advanced drug delivery. [Display omitted] [ABSTRACT FROM AUTHOR]

Technologies enabling high-cell-density growth are required for economical industrial production of most biotechnological products. However, the key factor limiting cell density in bioreactors is the availability of oxygen during the late phases of fermentation. Although the expression of bacterial Vitreoscilla hemoglobin (VHb) is useful for enhanced oxygen availability, bacterial cell membrane makes efficient hemoglobin-oxygen contact a challenge. On the other hand, periplasmic spaces of Gram-negative microorganisms offer an excellent compartment for the intermittent storage of hemoglobin-bound oxygen. In this study, the cell growth was increased by a remarkable 100% using the twin-arginine translocase (Tat) pathway to export active VHb into the periplasm of Escherichia coli, Halomonas bluephagenesis TD01 and H. campaniensis LS21. Furthermore, eight low-oxygen-inducible vgb promoters were constructed in tandem to become a strong promoter cassette termed P 8vgb , which better induces expression of both gene vgb encoding VHb and the PHB synthesis operon microaerobically. Both the P 8vgb and VHb performed excellently in E. coli and two Halomonas spp., demonstrating their universal applicability for various organisms. [ABSTRACT FROM AUTHOR]