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Journal articleZhou Y, Sun M-L, Lin L, et al., 2025,
Dynamic regulation combined with systematic metabolic engineering for high-level palmitoleic acid accumulation in oleaginous yeast.
, Metab Eng, Vol: 89, Pages: 33-46Palmitoleic acid (POA, C16:1Δ9) is widely recognized for its preventive and therapeutic effects in various chronic and cardiovascular diseases, but the current production practices based on plant extraction are both economically and ecologically unsustainable. Although Yarrowia lipolytica is capable of producing POA, it only accumulates to a small percentage of total fatty acids. The present study aimed to enhance the accumulation of POA by employing a two-layer engineering strategy, encompassing the modulation of the fatty acid profile and the promotion of the accumulation of POA-rich lipids. The fatty acid profile was subject to modulation through the engineering of the fatty acid metabolism by expressing heterologous specific fatty acid desaturases CeFat5 and implementing dynamic regulation based on a copper-responsive promoter. Then, the mechanism underlying this improvement of POA production capacity was elucidated. Finally, the POA-rich lipid accumulation ability was enhanced through engineering of the lipid metabolism by overexpressing the heterologous POA-specific triacylglycerol forming acyltransferase, introducing the artificial designed non-carboxylative malonyl-CoA production pathway, and preventing lipid degradation. The resulting optimized yeast strain achieved an impressive POA accumulation accounting for 50.62% of total fatty acids, marking a 37.7-fold improvement over the initial strain. Moreover, a record POA titer of 25.6 g/L was achieved in the bioreactor. Overall, this study introduces a framework for establishing efficient yeast platforms for the accumulation of valuable fatty acids.
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Journal articleÖztürk HB, Verdross P, Bismarck A, 2025,
Black liquor derived lignin adsorbents for removal of organic pollutants from water
, Reactive and Functional Polymers, Vol: 209, ISSN: 1381-5148We present the preparation of lignin particles from raw black liquor and their chemical modification, so that they can be used as efficient adsorbents to remove organic pollutants from water. For the production of adsorbent powders, pristine black liquor (as obtained from a pulping mill) was emulsified with epichlorohydrin and Span 80 and cured at elevated temperatures. Phospinic esters and organic sulphates were synthetically introduced to the lignin-backbone to modify the adsorbents. The adsorption of organic model compounds, Paraquat, p-nitrophenol, and malachite green were studied. Kinetic and thermodynamic data of the adsorption processes was obtained experimentally and by fitting mathematical models (Langmuir and Freundlich isotherms). Adsorbents showed adsorption capacities of Qe = 62 mg/g for Paraquat, Qe = 199 mg/g for p-nitrophenol, and Qe = 1200 mg/g for malachite green in batch-adsorption experiments. These values are among the highest for any of the model compounds presented in literature. Adsorption occurred reasonably fast for all compounds, reaching plateau-adsorption within minutes to hours.
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Journal articleHummer S, Barkan-Öztürk H, Bismarck A, et al., 2025,
Ready, steady, flow: Pd-loaded hypercrosslinked microreactors for the flow synthesis of paracetamol
, Applied Materials Today, Vol: 43, ISSN: 2352-9407We describe the implementation of monolithic polymerised high internal phase emulsions (polyHIPEs) as catalyst supports for the continuous flow synthesis of paracetamol. PolyHIPEs are formed by polymerisation of particle and surfactant-stabilised water-in-oil HIPEs comprising of a divinylbenzene and divinylbenzene/4-vinylbenzyl chloride monomer continuous phase. The polyHIPEs were post-functionalised by hypercrosslinking using the Scholl coupling reaction to yield polymers decorated with organophosphorus moieties and surface areas of up to 500 m²/g. The phosphorus moieties act as catalyst anchor sites for the subsequent loading of Pd via a Pd−O=P ligand bridge. The Pd-loaded polyHIPEs are used as catalysts for the reduction of 4-nitrophenol to 4-aminophenol, achieving turnover numbers of ∼2,000. The Pd-loaded polyHIPEs and non-functionalised divinylbenzene-based polyHIPEs are fabricated into flow-cells and used en route to Plant-on-a-Bench as microreactor/mixer for the continuous flow synthesis of paracetamol in yields of 66%.
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Journal articleYoun T, Kim G, Hariharan P, et al., 2025,
Improved Pendant-Bearing Glucose-Neopentyl Glycols for Membrane Protein Stability.
, Bioconjug ChemMembrane proteins are biologically and pharmaceutically significant, and determining their 3D structures requires a membrane-mimetic system to maintain protein stability. Detergent micelles are widely used as membrane mimetics; however, their dynamic structures often lead to the denaturation and aggregation of encapsulated membrane proteins. To address the limitations of classical detergents in stabilizing membrane proteins, we previously reported a class of glucose-neopentyl glycols (GNGs) and their pendant-bearing versions (P-GNGs), several of which proved more effective than DDM in stabilizing membrane proteins. In this study, we synthesized additional GNG derivatives by varying the lengths of the pendant (P-GNGs), and by introducing hemifluorinated pendants to the GNG scaffold (fluorinated pendant-bearing GNGs or FP-GNGs). The synthetic flexibility of the GNG chemical architecture allowed us to create a diverse range of derivatives, essential for the effective optimization of detergent properties. When tested with two model membrane proteins (a transporter and a G-protein coupled receptor (GPCR)), most of the new (F)P-GNGs demonstrated superior stabilization of these membrane proteins compared to DDM, the original GNG (OGNG)), and a previously developed P-GNG (i.e., GNG-3,14). Notably, several P-GNGs synthesized in this study were as effective as or even better than lauryl maltose neopentyl glycol (LMNG) in stabilizing a human GPCR, beta2 adrenergic receptor (β2AR). Enhanced protein stability was particularly observed for the P-GNGs with a butyl (C4) or pentyl (C5) pendant, indicating that these pendant sizes are optimal for membrane protein stability. The volumes of these pendants appear to minimize the empty spaces in the micelle interiors, thereby enhancing detergent-detergent interactions in micelles complexed with the membrane proteins. Additionally, we identified one FP-GNG that was more efficient at extracting the transporter and more effective at st
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Journal articleSadaf A, Yun HS, Lee H, et al., 2025,
Multiple Pendants-Bearing Triglucosides for Membrane Protein Studies: Effects of Pendant Length and Number on Micelle Interior Hydration and Protein Stability.
, BiomacromoleculesMembrane proteins play central roles in cell physiology and are the targets of over 50% of FDA-approved drugs. In the present study, we prepared single alkyl-chained triglucosides decorated with multiple pendants, designated multiple pendant-bearing glucosides (MPGs), to enhance membrane protein stability. The new detergents feature two and four pendants of varying size at the hydrophilic-lipophilic interfaces, designated MPG-Ds and MPG-Ts, respectively. When tested with model membrane proteins, including the human adrenergic receptor (β2AR), the tetra-pendant-bearing MPGs (MPG-Ts) demonstrated superior performance compared to the dipendant analogs (MPG-Ds) and the gold standard DDM. All-atom molecular dynamics (MD) simulations results reveal that the four-pendant configuration of this detergent is remarkably effective in excluding water from the hydrophobic micelle interiors compared to the dipendant MPGs and DDM, an unprecedented feature of this new detergent. Our findings provide a novel strategy for designing water-resistant detergents, advancing the field of membrane protein research.
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Journal articleDodds IL, Watts EC, Schuster M, et al., 2025,
Immunity gene silencing increases transient protein expression in Nicotiana benthamiana.
, Plant Biotechnol J -
Journal articleRawson TM, Al-Hassan M, Brzeska-Trafny I, et al., 2025,
Comment on: Resistance profiles of carbapenemase-producing Enterobacterales in a large centre in England: are we already losing cefiderocol?
, J Antimicrob Chemother -
Journal articleSun M-L, Xu Y, Lin L, et al., 2025,
Enhancing Precursor Supply and Engineering Efflux Systems to Improve Abscisic Acid Production and Secretion in Yarrowia lipolytica.
, J Agric Food Chem, Vol: 73, Pages: 6050-6058Abscisic acid is a sesquiterpene phytohormone with extensive applications in agriculture and human health. Currently, it is produced through fermentation of Botrytis cinerea, a plant pathogenic filamentous fungus. The process requires morphology controls, which complicates production and strain optimization. In this study, the abscisic acid production strain Yarrowia lipolytica SM309 was optimized by enhancing the precursor supply using a "push-pull-restrain" strategy focusing on acetyl-CoA, which increased abscisic acid production from 266.34 to 328.51 mg/L. Subsequently, in silico prediction and analysis were used to obtain the docking conformations and binding affinity of ABC transporters for abscisic acid. Overexpression of ABC transporter YlGcn20 further enhanced abscisic acid production by 10.88%, reaching 354.21 mg/L. Additionally, low temperature and dodecane addition were employed as auxiliary strategies to promote abscisic acid synthesis, resulting in a titer of 605.92 mg/L. Finally, the engineered strain achieved an abscisic acid titer of 2056.64 mg/L in a 5 L bioreactor, representing the highest titer reported for a yeast de novo synthesis system to date.
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Journal articleSamir NM, Locke-Gotel J, Urayama S-I, et al., 2025,
Molecular characterization of a polymycovirus in Leptosphaeria biglobosa.
, Arch Virol, Vol: 170Leptosphaeria biglobosa is a phytopathogenic ascomycete of Brassica napus that causes phoma stem canker/blackleg. A new double-stranded RNA (dsRNA) mycovirus from this fungus has been fully characterized. The virus genome has five dsRNA segments, ranging in length from 1,180 bp to 2,402 bp. Each dsRNA has a single open reading frame flanked by 5' and 3' untranslated regions. The proteins encoded by dsRNAs 1 and 3, an RNA-dependent RNA polymerase (RdRP) and a methyltransferase, respectively, have significant similarity to those of Plasmopara viticola lesion associated polymycovirus 1. The proline-alanine-serine-rich protein encoded by dsRNA 5 is similar to that of Erysiphe necator associated polymycovirus 1. The proteins encoded by dsRNAs 2 and 4 have significant similarity to those of a mycovirus identified in Alternaria sp. FA0703. Phylogenetic analysis based on RdRP sequences showed that this virus clusters with members of the family Polymycoviridae. Based on these observations, this virus, which we have named "Leptosphaeria biglobosa polymycovirus 1", should be classified as a member of the family Polymycoviridae. This is the first report of a polymycovirus in L. biglobosa.
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Journal articleZheng X, Li Y, Che C, et al., 2025,
Metabolic Engineering of Rhodotorula toruloides for the Production of Linalool.
, J Agric Food Chem, Vol: 73, Pages: 5395-5404Linalool is widely used in foods, pharmaceuticals, and cosmetics. Microbial production offers a sustainable alternative to current plant extraction and chemical synthesis. In the present study, the oleaginous yeast Rhodotorula toruloides was metabolically engineered for linalool production. Overall, an integrated strategy involving linalool synthase selection, promoter screening, and pathway enhancement was implemented. The linalool biosynthesis baseline was established by using a 51 aa-truncated linalool synthase from Mentha citrate (t51McLIS). The linalool titer was increased to 249.7 mg/L by optimizing the expression of critical enzymes with native promoters. Then, it was improved to 790.2 mg/L by manipulating the mevalonate pathway and further elevated to 845.1 mg/L (42.3 mg/g glucose) by optimizing the fermentation conditions in shake flasks. Finally, the linalool production was enhanced to 2.59 g/L and 26.9 mg/g glucose in a 3 L bioreactor. The results demonstrated the potential of R. toruloides for the microbial production of monoterpenes.
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