Pub Date : 2024-09-13DOI: 10.1038/s42004-024-01299-9
Kangning Liu, Qi Ding, Doudou Cao, Enpeng Xi, Yun Zhao, Nan Gao, Yajie Yang, Ye Yuan
Although the pathogenesis of Alzheimer’s disease (AD) is still unknown, the molecular pathological phenomena is clear, mainly due to mitochondrial dysfunction and central nervous system inflammation caused by imbalanced antioxidant capacity and synaptic dysfunction, so antioxidant therapy is still the preferred treatment for AD. However, although antioxidant enzymes have high catalytic efficiency, the substrate spectrum is narrow; Antioxidants have wider range of effects, but their efficiency is low. Since the antioxidant defense system in high-grade organisms is composed of both enzymatic and non-enzymatic systems, therefore we synthesized a metal-organic framework (MOF) with superoxide dismutase activity, and depending on the interface potential effect, curcumin was loaded to construct a synergistic antioxidant treatment system. More importantly, due to the complementary surface electrostatic potential between MOF and curcumin, the system exhibited both good antioxidant activity and efficient β-amyloid plaque scavenging ability, which slowed down the cognitive dysfunction in the brain of AD mice. Although the pathogenesis of Alzheimer’s disease (AD) is still unknown, imbalanced antioxidant capacity in nerve cells is a successfully targeted pathological phenomenon in clinical practice. Here, the authors show that the complementary surface electrostatic potential between a metal-organic framework and curcumin results in a complex with good antioxidant activity and efficient β-amyloid plaque scavenging ability, which slows down the cognitive dysfunction in the brain of AD mice.
{"title":"Interface potential-induced natural antioxidant mimic system for the treatment of Alzheimer’s disease","authors":"Kangning Liu, Qi Ding, Doudou Cao, Enpeng Xi, Yun Zhao, Nan Gao, Yajie Yang, Ye Yuan","doi":"10.1038/s42004-024-01299-9","DOIUrl":"10.1038/s42004-024-01299-9","url":null,"abstract":"Although the pathogenesis of Alzheimer’s disease (AD) is still unknown, the molecular pathological phenomena is clear, mainly due to mitochondrial dysfunction and central nervous system inflammation caused by imbalanced antioxidant capacity and synaptic dysfunction, so antioxidant therapy is still the preferred treatment for AD. However, although antioxidant enzymes have high catalytic efficiency, the substrate spectrum is narrow; Antioxidants have wider range of effects, but their efficiency is low. Since the antioxidant defense system in high-grade organisms is composed of both enzymatic and non-enzymatic systems, therefore we synthesized a metal-organic framework (MOF) with superoxide dismutase activity, and depending on the interface potential effect, curcumin was loaded to construct a synergistic antioxidant treatment system. More importantly, due to the complementary surface electrostatic potential between MOF and curcumin, the system exhibited both good antioxidant activity and efficient β-amyloid plaque scavenging ability, which slowed down the cognitive dysfunction in the brain of AD mice. Although the pathogenesis of Alzheimer’s disease (AD) is still unknown, imbalanced antioxidant capacity in nerve cells is a successfully targeted pathological phenomenon in clinical practice. Here, the authors show that the complementary surface electrostatic potential between a metal-organic framework and curcumin results in a complex with good antioxidant activity and efficient β-amyloid plaque scavenging ability, which slows down the cognitive dysfunction in the brain of AD mice.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-6"},"PeriodicalIF":5.9,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01299-9.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142233247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1038/s42004-024-01296-y
Krishna Puri, Suvarn S. Kulkarni
Zwitterionic polysaccharides (ZPSs) present on the surface of a common gut commensal Bacteroides fragilis are endowed with unique immunological properties as they can directly bind to T-cells in the absence of protein conjugation. ZPSs are therefore considered to be potential antigens for the development of totally carbohydrate-based vaccines. Herein, we disclose the first total synthesis of a highly branched phosphorylated zwitterionic capsular polysaccharide repeating unit of Bacteroides fragilis. The hexasaccharide repeating unit bearing six different monosaccharides comprises three 1,2-cis-glycosidic linkages, a challenging 1,2-trans linkage in D-QuipNAc-β-(1→4)-D-Gal motif, and a 2-aminoethyl phosphonate appendage. The synthesis of target ZPS was accomplished utilizing an expeditious, highly stereoselective and convergent (1 + 2 + 2 + 1) one-pot glycosylation strategy. The striking features include efficient synthesis of rare deoxy amino sugars D- and L-quinovosamine, stereoselective installation of three 1,2-cis glycosidic linkages, glycosylation of D-quinovosamine donor with a sterically crowded, poorly reactive 4-OH galactose moiety, as well as late stage phosphorylation. Zwitterionic polysaccharides present on the surface of a common gut commensal Bacteroides fragilis are considered to be potential antigens for the development of totally carbohydrate-based vaccines. Here, the authors report the total synthesis of a highly branched phosphorylated zwitterionic capsular hexasaccharide repeating unit of Bacteroides fragilis via a one-pot glycosylation strategy.
{"title":"Total synthesis of a structurally complex zwitterionic hexasaccharide repeating unit of polysaccharide B from Bacteroides fragilis via one-pot glycosylation","authors":"Krishna Puri, Suvarn S. Kulkarni","doi":"10.1038/s42004-024-01296-y","DOIUrl":"10.1038/s42004-024-01296-y","url":null,"abstract":"Zwitterionic polysaccharides (ZPSs) present on the surface of a common gut commensal Bacteroides fragilis are endowed with unique immunological properties as they can directly bind to T-cells in the absence of protein conjugation. ZPSs are therefore considered to be potential antigens for the development of totally carbohydrate-based vaccines. Herein, we disclose the first total synthesis of a highly branched phosphorylated zwitterionic capsular polysaccharide repeating unit of Bacteroides fragilis. The hexasaccharide repeating unit bearing six different monosaccharides comprises three 1,2-cis-glycosidic linkages, a challenging 1,2-trans linkage in D-QuipNAc-β-(1→4)-D-Gal motif, and a 2-aminoethyl phosphonate appendage. The synthesis of target ZPS was accomplished utilizing an expeditious, highly stereoselective and convergent (1 + 2 + 2 + 1) one-pot glycosylation strategy. The striking features include efficient synthesis of rare deoxy amino sugars D- and L-quinovosamine, stereoselective installation of three 1,2-cis glycosidic linkages, glycosylation of D-quinovosamine donor with a sterically crowded, poorly reactive 4-OH galactose moiety, as well as late stage phosphorylation. Zwitterionic polysaccharides present on the surface of a common gut commensal Bacteroides fragilis are considered to be potential antigens for the development of totally carbohydrate-based vaccines. Here, the authors report the total synthesis of a highly branched phosphorylated zwitterionic capsular hexasaccharide repeating unit of Bacteroides fragilis via a one-pot glycosylation strategy.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-15"},"PeriodicalIF":5.9,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01296-y.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142236064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1038/s42004-024-01297-x
Isabella Senini, Sara Tengattini, Francesca Rinaldi, Gabriella Massolini, Christoph Gstöttner, Dietmar Reusch, Marcello Donini, Carla Marusic, Peter A. van Veelen, Elena Domínguez-Vega, Manfred Wuhrer, Caterina Temporini, Simone Nicolardi
Monoclonal antibody (mAb) glycoengineering has the potential to improve the efficacy of biopharmaceuticals by fine-tuning specific biological properties. Glycosylation analysis is key to monitoring the glycoengineering process. Various mass spectrometry (MS)-based methods are available to characterize mAb glycosylation at different structural levels, but comprehensive analysis is typically time-consuming and costly. Here, we present an approach that combines conventional intact mass measurement of glycoforms by direct infusion ESI-MS with an advanced MALDI-in-source decay FT-ICR MS method for direct analysis of glycans in intact mAbs, without the need for enzymatic release and separation. Using a sodium-doped MALDI matrix, glycans were directly released as ISD fragment ions from the intact mAbs during the ionization process. Measurement of 0,2A fragment signals yielded reproducible glycan profiles that were consistent with conventional methods, yet was achieved with unprecedented speed, providing complementary information to that obtained through intact mass measurement. The methods were applied to standard and glycoengineered trastuzumab and rituximab, allowing rapid glycosylation profiling and structural analysis of glycans by tandem MS of selected ISD fragment ions. This fast approach can facilitate the early-phase development of glycoengineering processes by constraining further in-depth analyses. We envision a broader applicability in studies focused on glycosylation changes in mAbs. Glycoengineering of monoclonal antibodies (mAbs) has the potential to improve the efficacy of biopharmaceuticals, however, monitoring the glycoengineering process by glycosylation analysis often requires a multi-method approach. Here, the authors present a direct glycosylation analysis of intact mAbs by combining conventional ESI-MS of intact glycoforms and MALDI-in-source decay FT-ICR MS of glycan fragments.
单克隆抗体（mAb）糖工程有可能通过微调特定的生物特性来提高生物制药的疗效。糖基化分析是监测糖工程过程的关键。目前有多种基于质谱（MS）的方法来表征不同结构水平的 mAb 糖基化，但全面的分析通常耗时且成本高昂。在这里，我们提出了一种方法，它将传统的直接注入 ESI-MS 测量糖型的完整质量测量方法与先进的 MALDI-in-source decay FT-ICR MS 方法相结合，直接分析完整 mAb 中的聚糖，而无需酶解和分离。使用掺钠 MALDI 基质，在离子化过程中，聚糖作为 ISD 片段离子从完整的 mAbs 中直接释放出来。对 0,2A 片段信号的测量得到了与传统方法一致的可重现的聚糖图谱，而且以前所未有的速度完成，提供了与通过完整质量测量获得的信息互补的信息。这些方法被应用于标准曲妥珠单抗和利妥昔单抗以及经过糖工程化的曲妥珠单抗和利妥昔单抗，从而可以通过对选定的 ISD 片段离子进行串联质谱分析来快速绘制糖基化图谱和分析聚糖的结构。这种快速方法可以限制进一步的深入分析，从而促进糖工程过程的早期开发。我们设想这种方法将更广泛地应用于以 mAbs 糖基化变化为重点的研究中。
{"title":"Direct glycosylation analysis of intact monoclonal antibodies combining ESI MS of glycoforms and MALDI-in source decay MS of glycan fragments","authors":"Isabella Senini, Sara Tengattini, Francesca Rinaldi, Gabriella Massolini, Christoph Gstöttner, Dietmar Reusch, Marcello Donini, Carla Marusic, Peter A. van Veelen, Elena Domínguez-Vega, Manfred Wuhrer, Caterina Temporini, Simone Nicolardi","doi":"10.1038/s42004-024-01297-x","DOIUrl":"10.1038/s42004-024-01297-x","url":null,"abstract":"Monoclonal antibody (mAb) glycoengineering has the potential to improve the efficacy of biopharmaceuticals by fine-tuning specific biological properties. Glycosylation analysis is key to monitoring the glycoengineering process. Various mass spectrometry (MS)-based methods are available to characterize mAb glycosylation at different structural levels, but comprehensive analysis is typically time-consuming and costly. Here, we present an approach that combines conventional intact mass measurement of glycoforms by direct infusion ESI-MS with an advanced MALDI-in-source decay FT-ICR MS method for direct analysis of glycans in intact mAbs, without the need for enzymatic release and separation. Using a sodium-doped MALDI matrix, glycans were directly released as ISD fragment ions from the intact mAbs during the ionization process. Measurement of 0,2A fragment signals yielded reproducible glycan profiles that were consistent with conventional methods, yet was achieved with unprecedented speed, providing complementary information to that obtained through intact mass measurement. The methods were applied to standard and glycoengineered trastuzumab and rituximab, allowing rapid glycosylation profiling and structural analysis of glycans by tandem MS of selected ISD fragment ions. This fast approach can facilitate the early-phase development of glycoengineering processes by constraining further in-depth analyses. We envision a broader applicability in studies focused on glycosylation changes in mAbs. Glycoengineering of monoclonal antibodies (mAbs) has the potential to improve the efficacy of biopharmaceuticals, however, monitoring the glycoengineering process by glycosylation analysis often requires a multi-method approach. Here, the authors present a direct glycosylation analysis of intact mAbs by combining conventional ESI-MS of intact glycoforms and MALDI-in-source decay FT-ICR MS of glycan fragments.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-10"},"PeriodicalIF":5.9,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01297-x.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142210250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-09DOI: 10.1038/s42004-024-01279-z
Sanfeng Dong, Hui Huang, Jintian Li, Xiaomei Li, Samuel Jacob Bunu, Yun Yang, Yong Zhang, Qi Jia, Zhijian Xu, Yingxia Li, Hu Zhou, Bo Li, Weiliang Zhu
Multi-functional cysteine-targeting covalent warheads possess significant therapeutic potential in medicinal chemistry and chemical biology. Herein, we present novel unsaturated and asymmetric ketone (oxazolinosene) scaffolds that selectively conjugate cysteine residues of peptides and bovine serum albumin under normal physiological conditions. This unsaturated saccharide depletes GSH in NCI-H1299 cells, leading to anti-tumor effects in vitro. The acetyl group of the ketal moiety on the saccharide ring can be converted to other carboxylic acids in a one-pot synthesis. In this way, the loaded acid can be click-released during cysteine conjugation, making the oxazolinosene a potential multifunctional therapeutic agent. The reaction kinetic model for oxazolinosene conjugation to GSH is well established and was used to evaluate oxazolinosene reactivity. The aforementioned oxazolinosenes were stereoselectively synthesized via a one-step reaction of nitriles with saccharides and conveniently converted into a series of α, β-unsaturated ketone N-glycosides as prevalent synthetic building blocks. The reaction mechanisms of oxazolinosene synthesis were investigated through calculations and validated with control experiments. Overall, these oxazolinosenes can be easily synthesized and developed as cysteine-targeted covalent warheads carrying useful click-releasing groups. Multifunctional cysteine targeting covalent warheads possess significant therapeutic potential in medicinal chemistry and chemical biology. Here, the authors develop an oxazolinosene scaffold from nitrile groups and saccharides that can selectively conjugate cysteine residues within peptides and proteins under physiological conditions, as well as deplete glutathione in cancer cells.
{"title":"Development of ketalized unsaturated saccharides as multifunctional cysteine-targeting covalent warheads","authors":"Sanfeng Dong, Hui Huang, Jintian Li, Xiaomei Li, Samuel Jacob Bunu, Yun Yang, Yong Zhang, Qi Jia, Zhijian Xu, Yingxia Li, Hu Zhou, Bo Li, Weiliang Zhu","doi":"10.1038/s42004-024-01279-z","DOIUrl":"10.1038/s42004-024-01279-z","url":null,"abstract":"Multi-functional cysteine-targeting covalent warheads possess significant therapeutic potential in medicinal chemistry and chemical biology. Herein, we present novel unsaturated and asymmetric ketone (oxazolinosene) scaffolds that selectively conjugate cysteine residues of peptides and bovine serum albumin under normal physiological conditions. This unsaturated saccharide depletes GSH in NCI-H1299 cells, leading to anti-tumor effects in vitro. The acetyl group of the ketal moiety on the saccharide ring can be converted to other carboxylic acids in a one-pot synthesis. In this way, the loaded acid can be click-released during cysteine conjugation, making the oxazolinosene a potential multifunctional therapeutic agent. The reaction kinetic model for oxazolinosene conjugation to GSH is well established and was used to evaluate oxazolinosene reactivity. The aforementioned oxazolinosenes were stereoselectively synthesized via a one-step reaction of nitriles with saccharides and conveniently converted into a series of α, β-unsaturated ketone N-glycosides as prevalent synthetic building blocks. The reaction mechanisms of oxazolinosene synthesis were investigated through calculations and validated with control experiments. Overall, these oxazolinosenes can be easily synthesized and developed as cysteine-targeted covalent warheads carrying useful click-releasing groups. Multifunctional cysteine targeting covalent warheads possess significant therapeutic potential in medicinal chemistry and chemical biology. Here, the authors develop an oxazolinosene scaffold from nitrile groups and saccharides that can selectively conjugate cysteine residues within peptides and proteins under physiological conditions, as well as deplete glutathione in cancer cells.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-10"},"PeriodicalIF":5.9,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01279-z.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142160319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Although luminescent aluminum compounds have been utilized for emitting and electron transporting layers in organic light-emitting diodes, most of them often exhibit not phosphorescence but fluorescence with lower photoluminescent quantum yields in the aggregated state than those in the amorphous state due to concentration quenching. Here we show the synthesis and optical properties of β-diketiminate aluminum complexes, such as crystallization-induced emission (CIE) and room-temperature phosphorescence (RTP), and the substituent effects of the central element. The dihaloaluminum complexes were found to exhibit the CIE property, especially RTP from the diiodo complex, while the dialkyl ones showed almost no emission in both solution and solid states. Theoretical calculations suggested that undesired structural relaxation in the singlet excited state of dialkyl complexes should be suppressed by introducing electronegative halogens instead of alkyl groups. Our findings could provide a molecular design not only for obtaining luminescent complexes but also for achieving triplet-harvesting materials. Luminescent aluminum compounds have been utilized for emitting and electron transporting layers in organic light-emitting diodes, but most exhibit fluorescence as opposed to phosphorescence. Here, the photophysical properties of β-diketiminate aluminum complexes are shown to depend on the nature of the metal substituents, with a diiodoaluminum complex displaying room temperature phosphorescence.
{"title":"Regulating the photoluminescence of aluminium complexes from non-luminescence to room-temperature phosphorescence by tuning the metal substituents","authors":"Shunichiro Ito, Takuya Hosokai, Kazuo Tanaka, Yoshiki Chujo","doi":"10.1038/s42004-024-01295-z","DOIUrl":"10.1038/s42004-024-01295-z","url":null,"abstract":"Although luminescent aluminum compounds have been utilized for emitting and electron transporting layers in organic light-emitting diodes, most of them often exhibit not phosphorescence but fluorescence with lower photoluminescent quantum yields in the aggregated state than those in the amorphous state due to concentration quenching. Here we show the synthesis and optical properties of β-diketiminate aluminum complexes, such as crystallization-induced emission (CIE) and room-temperature phosphorescence (RTP), and the substituent effects of the central element. The dihaloaluminum complexes were found to exhibit the CIE property, especially RTP from the diiodo complex, while the dialkyl ones showed almost no emission in both solution and solid states. Theoretical calculations suggested that undesired structural relaxation in the singlet excited state of dialkyl complexes should be suppressed by introducing electronegative halogens instead of alkyl groups. Our findings could provide a molecular design not only for obtaining luminescent complexes but also for achieving triplet-harvesting materials. Luminescent aluminum compounds have been utilized for emitting and electron transporting layers in organic light-emitting diodes, but most exhibit fluorescence as opposed to phosphorescence. Here, the photophysical properties of β-diketiminate aluminum complexes are shown to depend on the nature of the metal substituents, with a diiodoaluminum complex displaying room temperature phosphorescence.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-9"},"PeriodicalIF":5.9,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01295-z.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142160258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-07DOI: 10.1038/s42004-024-01288-y
Guiyeoul Lim, Donato Calabrese, Allison Wolder, Paul R. F. Cordero, Dörte Rother, Florian F. Mulks, Caroline E. Paul, Lars Lauterbach
Despite the increasing demand for efficient and sustainable chemical processes, the development of scalable systems using biocatalysis for fine chemical production remains a significant challenge. We have developed a scalable flow system using immobilized enzymes to facilitate flavin-dependent biocatalysis, targeting as a proof-of-concept asymmetric alkene reduction. The system integrates a flavin-dependent Old Yellow Enzyme (OYE) and a soluble hydrogenase to enable H2-driven regeneration of the OYE cofactor FMNH2. Molecular hydrogen was produced by water electrolysis using a proton exchange membrane (PEM) electrolyzer and introduced into the flow system via a designed gas membrane addition module at a high diffusion rate. The flow system shows remarkable stability and reusability, consistently achieving >99% conversion of ketoisophorone to levodione. It also demonstrates versatility and selectivity in reducing various cyclic enones and can be extended to further flavin-based biocatalytic approaches and gas-dependent reactions. This electro-driven continuous flow system, therefore, has significant potential for advancing sustainable processes in fine chemical synthesis. Flavin-based biocatalysis using flavin mononucleotide (FMN) cofactor attracts significant attention for its application in asymmetric alkene reduction and various other reactions, however, the scale-up of flavin-based biocatalysis in flow remains unexplored. Here, the authors develop a closed-loop flow platform for H2-driven regeneration of cofactor FMNH2 and ene-reduction using immobilized Old Yellow Enzyme, achieving >99% conversion of ketoisophorone to levodione.
{"title":"H2-driven biocatalysis for flavin-dependent ene-reduction in a continuous closed-loop flow system utilizing H2 from water electrolysis","authors":"Guiyeoul Lim, Donato Calabrese, Allison Wolder, Paul R. F. Cordero, Dörte Rother, Florian F. Mulks, Caroline E. Paul, Lars Lauterbach","doi":"10.1038/s42004-024-01288-y","DOIUrl":"10.1038/s42004-024-01288-y","url":null,"abstract":"Despite the increasing demand for efficient and sustainable chemical processes, the development of scalable systems using biocatalysis for fine chemical production remains a significant challenge. We have developed a scalable flow system using immobilized enzymes to facilitate flavin-dependent biocatalysis, targeting as a proof-of-concept asymmetric alkene reduction. The system integrates a flavin-dependent Old Yellow Enzyme (OYE) and a soluble hydrogenase to enable H2-driven regeneration of the OYE cofactor FMNH2. Molecular hydrogen was produced by water electrolysis using a proton exchange membrane (PEM) electrolyzer and introduced into the flow system via a designed gas membrane addition module at a high diffusion rate. The flow system shows remarkable stability and reusability, consistently achieving >99% conversion of ketoisophorone to levodione. It also demonstrates versatility and selectivity in reducing various cyclic enones and can be extended to further flavin-based biocatalytic approaches and gas-dependent reactions. This electro-driven continuous flow system, therefore, has significant potential for advancing sustainable processes in fine chemical synthesis. Flavin-based biocatalysis using flavin mononucleotide (FMN) cofactor attracts significant attention for its application in asymmetric alkene reduction and various other reactions, however, the scale-up of flavin-based biocatalysis in flow remains unexplored. Here, the authors develop a closed-loop flow platform for H2-driven regeneration of cofactor FMNH2 and ene-reduction using immobilized Old Yellow Enzyme, achieving >99% conversion of ketoisophorone to levodione.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-7"},"PeriodicalIF":5.9,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01288-y.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142145335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-04DOI: 10.1038/s42004-024-01270-8
Chang Chen, Ketan A. Ganar, Robbert J. de Haas, Nele Jarnot, Erwin Hogeveen, Renko de Vries, Siddharth Deshpande
Compartmentalization is a vital aspect of living cells to orchestrate intracellular processes. In a similar vein, constructing dynamic and responsive sub-compartments is key to synthetic cell engineering. In recent years, liquid-liquid phase separation via coacervation has offered an innovative avenue for creating membraneless organelles (MOs) within artificial cells. Here, we present a lab-on-a-chip system to reversibly trigger peptide-based coacervates within cell-mimicking confinements. We use double emulsion droplets (DEs) as our synthetic cell containers while pH-responsive elastin-like polypeptides (ELPs) act as the coacervate system. We first present a high-throughput microfluidic DE production enabling efficient encapsulation of the ELPs. The DEs are then harvested to perform multiple MO formation-dissolution cycles using pH as well as temperature variation. For controlled long-term visualization and modulation of the external environment, we developed an integrated microfluidic device for trapping and environmental stimulation of DEs, with negligible mechanical force, and demonstrated a proof-of-principle osmolyte-based triggering to induce multiple MO formation-dissolution cycles. In conclusion, our work showcases the use of DEs and ELPs in designing membraneless reversible compartmentalization within synthetic cells via physicochemical triggers. Additionally, presented on-chip platform can be applied over a wide range of phase separation and vesicle systems for applications in synthetic cells and beyond. Compartmentalization within living cells is vital to orchestrate intracellular processes, but effective compartmentalization and organization within synthetic cells remains a key challenge. Here, the authors report a lab-on-a-chip system to reversibly trigger the formation of peptide-based coacervates as membraneless organelles via pH/temperature/osmolyte variations within cell-mimicking confinements.
区隔化是活细胞协调细胞内过程的一个重要方面。同样，构建动态和反应灵敏的亚细胞器也是合成细胞工程的关键。近年来，通过共凝的液-液相分离为在人造细胞内创建无膜细胞器（MO）提供了一条创新途径。在这里，我们展示了一种片上实验室系统，用于在模拟细胞的限制条件下可逆地触发基于肽的共凝物。我们使用双乳液液滴（DEs）作为合成细胞容器，而 pH 响应型弹性蛋白样多肽（ELPs）则作为凝聚剂系统。我们首先介绍了一种高通量微流控双乳液滴生产技术，它能有效地封装 ELPs。然后收获 DE，利用 pH 值和温度变化执行多个 MO 形成-溶解循环。为了实现可控的长期可视化和外部环境调控，我们开发了一种用于捕获和环境刺激 DEs 的集成微流控装置，其机械力可以忽略不计，并展示了一种基于渗透剂的触发原理验证，可诱导多个 MO 形成-溶解循环。总之，我们的工作展示了如何利用 DEs 和 ELPs 通过物理化学触发器在合成细胞内设计无膜可逆区隔。此外，所展示的片上平台可广泛应用于合成细胞内外的相分离和囊泡系统。活细胞内的区隔对于协调细胞内过程至关重要，但合成细胞内有效的区隔和组织仍然是一个关键挑战。在此，作者报告了一种片上实验室系统，该系统在模拟细胞的限制条件下，通过 pH 值/温度/溶质的变化，可逆地触发基于肽的凝聚态形成无膜细胞器。
{"title":"Elastin-like polypeptide coacervates as reversibly triggerable compartments for synthetic cells","authors":"Chang Chen, Ketan A. Ganar, Robbert J. de Haas, Nele Jarnot, Erwin Hogeveen, Renko de Vries, Siddharth Deshpande","doi":"10.1038/s42004-024-01270-8","DOIUrl":"10.1038/s42004-024-01270-8","url":null,"abstract":"Compartmentalization is a vital aspect of living cells to orchestrate intracellular processes. In a similar vein, constructing dynamic and responsive sub-compartments is key to synthetic cell engineering. In recent years, liquid-liquid phase separation via coacervation has offered an innovative avenue for creating membraneless organelles (MOs) within artificial cells. Here, we present a lab-on-a-chip system to reversibly trigger peptide-based coacervates within cell-mimicking confinements. We use double emulsion droplets (DEs) as our synthetic cell containers while pH-responsive elastin-like polypeptides (ELPs) act as the coacervate system. We first present a high-throughput microfluidic DE production enabling efficient encapsulation of the ELPs. The DEs are then harvested to perform multiple MO formation-dissolution cycles using pH as well as temperature variation. For controlled long-term visualization and modulation of the external environment, we developed an integrated microfluidic device for trapping and environmental stimulation of DEs, with negligible mechanical force, and demonstrated a proof-of-principle osmolyte-based triggering to induce multiple MO formation-dissolution cycles. In conclusion, our work showcases the use of DEs and ELPs in designing membraneless reversible compartmentalization within synthetic cells via physicochemical triggers. Additionally, presented on-chip platform can be applied over a wide range of phase separation and vesicle systems for applications in synthetic cells and beyond. Compartmentalization within living cells is vital to orchestrate intracellular processes, but effective compartmentalization and organization within synthetic cells remains a key challenge. Here, the authors report a lab-on-a-chip system to reversibly trigger the formation of peptide-based coacervates as membraneless organelles via pH/temperature/osmolyte variations within cell-mimicking confinements.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-11"},"PeriodicalIF":5.9,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01270-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142130454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-04DOI: 10.1038/s42004-024-01285-1
Hsiu-Wen Wang, Emily T. Nienhuis, Trent R. Graham, Maxime Pouvreau, Jacob G. Reynolds, Mark Bowden, Gregory K. Schenter, James J. De Yoreo, Kevin M. Rosso, Carolyn I. Pearce
Aluminum hydroxide polymorphs are of widespread importance yet their kinetics of nucleation and growth remain beyond the reach of current models. Here we attempt to unveil the reaction processes underlying the polymorphs formation at high chemical potential. We examine their formation in-situ from supersaturated alkaline sodium aluminate solutions using deuteration and time-resolved neutron pair distribution function analyses, which indicate the formation of individual Al(OD)3 layers as an intermediate particle phase. These layers ultimately stack to form gibbsite- or bayerite-like layered heterostructures. Ex-situ characterization of the recovered precipitates using 27Al magic angle spinning nuclear magnetic resonance spectroscopy, Raman, X-ray diffraction, and scanning electron microscopy, suggests the presence of additional intermediate states, an amorphous compound bearing both tetrahededrally- and penta-coordinated Al3+. These observations reveal the complex pathways to form Al(OD)3 monolayers via either transient pentacoordinate species or amorphous-to-ordered transitions. The subsequent crystallization of admixed gibbsite/bayerite is followed by an Al(OD)3 monolayer attachment process. Aluminum hydroxide polymorphs play a key role in industrial aluminum production, yet their nucleation and growth kinetics remain beyond the reach of current models. Here, the authors study polymorph formation in situ from supersaturated alkaline sodium aluminate solutions using time-resolved neutron pair distribution function and complementary spectroscopy analyses, which indicate the formation of individual Al(OD)3 layers as an intermediate particle phase.
{"title":"Resolving intermediates during the growth of aluminum deuteroxide (Hydroxide) polymorphs in high chemical potential solutions","authors":"Hsiu-Wen Wang, Emily T. Nienhuis, Trent R. Graham, Maxime Pouvreau, Jacob G. Reynolds, Mark Bowden, Gregory K. Schenter, James J. De Yoreo, Kevin M. Rosso, Carolyn I. Pearce","doi":"10.1038/s42004-024-01285-1","DOIUrl":"10.1038/s42004-024-01285-1","url":null,"abstract":"Aluminum hydroxide polymorphs are of widespread importance yet their kinetics of nucleation and growth remain beyond the reach of current models. Here we attempt to unveil the reaction processes underlying the polymorphs formation at high chemical potential. We examine their formation in-situ from supersaturated alkaline sodium aluminate solutions using deuteration and time-resolved neutron pair distribution function analyses, which indicate the formation of individual Al(OD)3 layers as an intermediate particle phase. These layers ultimately stack to form gibbsite- or bayerite-like layered heterostructures. Ex-situ characterization of the recovered precipitates using 27Al magic angle spinning nuclear magnetic resonance spectroscopy, Raman, X-ray diffraction, and scanning electron microscopy, suggests the presence of additional intermediate states, an amorphous compound bearing both tetrahededrally- and penta-coordinated Al3+. These observations reveal the complex pathways to form Al(OD)3 monolayers via either transient pentacoordinate species or amorphous-to-ordered transitions. The subsequent crystallization of admixed gibbsite/bayerite is followed by an Al(OD)3 monolayer attachment process. Aluminum hydroxide polymorphs play a key role in industrial aluminum production, yet their nucleation and growth kinetics remain beyond the reach of current models. Here, the authors study polymorph formation in situ from supersaturated alkaline sodium aluminate solutions using time-resolved neutron pair distribution function and complementary spectroscopy analyses, which indicate the formation of individual Al(OD)3 layers as an intermediate particle phase.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-10"},"PeriodicalIF":5.9,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01285-1.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142130455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-04DOI: 10.1038/s42004-024-01283-3
Samuel Mckeating, Oliver B. Penrhyn-Lowe, Sean Flynn, Savannah R. Cassin, Sarah Lomas, Christopher Fidge, Paul Price, Stephen Wright, Pierre Chambon, Steve P. Rannard
With the ever-growing reliance on polymeric materials for numerous applications, new avenues to induce, design and control degradation are clearly important. Here, we describe a previously unreported approach to controlling enzymatic hydrolysis of high molecular weight branched polymers formed from the new free-radical polymer synthesis strategy transfer-dominated branching radical telomerisation (TBRT). Modifying the chemical nature of TBRT polymers may be accomplished through telogen selection and multi-vinyl taxogen (MVT) design, and we show telogen-driven control of enzyme-catalysed hydrolysis and the impact of careful placement of hydrolytically susceptible groups within readily synthesised MVTs. Our results indicate that utilising conventional free-radical chemistries and unsaturated monomers as feedstocks for highly branched polymer architectures has considerable potential for the design of future materials that degrade into very low molecular weight byproducts at variable and controllable rates. With the ever-growing reliance on polymeric materials for numerous applications, new avenues to induce, design and control degradation are clearly important. Here, the authors report an approach to controlling the enzymatic hydrolysis of high molecular weight branched polymers formed from transfer-dominated branching radical telomerisation, through telogen selection and multi-vinyl taxogen design.
{"title":"Controlling enzyme hydrolysis of branched polymers synthesised using transfer-dominated branching radical telomerisation via telogen and taxogen selection","authors":"Samuel Mckeating, Oliver B. Penrhyn-Lowe, Sean Flynn, Savannah R. Cassin, Sarah Lomas, Christopher Fidge, Paul Price, Stephen Wright, Pierre Chambon, Steve P. Rannard","doi":"10.1038/s42004-024-01283-3","DOIUrl":"10.1038/s42004-024-01283-3","url":null,"abstract":"With the ever-growing reliance on polymeric materials for numerous applications, new avenues to induce, design and control degradation are clearly important. Here, we describe a previously unreported approach to controlling enzymatic hydrolysis of high molecular weight branched polymers formed from the new free-radical polymer synthesis strategy transfer-dominated branching radical telomerisation (TBRT). Modifying the chemical nature of TBRT polymers may be accomplished through telogen selection and multi-vinyl taxogen (MVT) design, and we show telogen-driven control of enzyme-catalysed hydrolysis and the impact of careful placement of hydrolytically susceptible groups within readily synthesised MVTs. Our results indicate that utilising conventional free-radical chemistries and unsaturated monomers as feedstocks for highly branched polymer architectures has considerable potential for the design of future materials that degrade into very low molecular weight byproducts at variable and controllable rates. With the ever-growing reliance on polymeric materials for numerous applications, new avenues to induce, design and control degradation are clearly important. Here, the authors report an approach to controlling the enzymatic hydrolysis of high molecular weight branched polymers formed from transfer-dominated branching radical telomerisation, through telogen selection and multi-vinyl taxogen design.","PeriodicalId":10529,"journal":{"name":"Communications Chemistry","volume":" ","pages":"1-11"},"PeriodicalIF":5.9,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.com/articles/s42004-024-01283-3.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142125077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-03DOI: 10.1038/s42004-024-01272-6
Mayukh Kansari, Fathia Idiris, Hendrik Szurmant, Tomáš Kubař, Alexander Schug
Histidine kinases (HK) are one of the main prokaryotic signaling systems. Two structurally conserved catalytic domains inside the HK enable autokinase, phosphotransfer, and phosphatase activities. Here, we focus on a detailed mechanistic understanding of the functional cycle of the WalK HK by a multi-scale simulation approach, consisting of classical as well as hybrid QM/MM molecular dynamics simulation. Strikingly, a conformational transition induced solely in DHp leads to the correct activated conformation in CA crucial for autophosphorylation. This finding explains how variable sensor domains induce the transition from inactive to active state. The subsequent autophosphorylation inside DHp proceeds via a penta-coordinated transition state to a protonated phosphohistidine intermediate. This intermediate is consequently deprotonated by a suitable nearby base. The reaction energetics are controlled by the final proton acceptor and presence of a magnesium cation. The slow rates of the process result from the high energy barrier of the conformational transition between inactive and active states. The phosphorylation step exhibits a lower barrier and down-the-hill energetics. Thus, our work suggests a detailed mechanistic model for HK autophosphorylation. Histidine kinases (HK) are the main component of a wide-spread signal transduction system in bacteria that are essential for cell viability, however, the details of HK autophosphorylation remain poorly understood. Here, the authors utilize a multi-scale simulation approach to investigate the mechanism of activation and autophosphorylation process, revealing the rate determining step and reaction free energy of the process.
组氨酸激酶（HK）是原核生物的主要信号系统之一。HK内部有两个结构一致的催化结构域，可实现自激酶、磷酸转移酶和磷酸酶活性。在这里，我们通过经典以及 QM/MM 混合分子动力学模拟等多尺度模拟方法，重点从机理上详细了解了 WalK HK 的功能循环。令人震惊的是，仅在 DHp 中诱导的构象转变导致 CA 中正确的激活构象，这对自动磷酸化至关重要。这一发现解释了可变传感器结构域如何诱导从非活性状态到活性状态的转变。随后，DHp 内部的自动磷酸化通过五配位过渡态进入质子化的磷组氨酸中间体。该中间体随后被附近合适的碱基去质子化。反应能量受最终质子接受体和镁阳离子的控制。由于非活性态和活性态之间构象转变的能垒较高，因此该过程的速率较慢。而磷酸化步骤则表现出较低的能量势垒和下坡能量。因此，我们的工作为 HK 自磷酸化提供了一个详细的机理模型。
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