Chiral Production Research Articles

Adaptive evolution of Paecilomyces variotii enhanced the biodetoxification of high-titer inhibitors in pretreated lignocellulosic feedstock

High inhibitor concentrations in lignocellulose feedstock negatively affect the degradation rate of biodetoxification strains. This study designed two adaptive laboratory evolutions in solid substrate and liquid medium to boost the biodetoxification capacity of P. variotii to high titers of lignocellulose-derived inhibitors, resulting in two evolved strains AC70 and ZW70. The results showed that the evolutionary adaptation in liquid medium could better boost the acetic acid assimilation compared to that on solid substrate. Transcriptional analysis revealed that the evolved strains exhibited a significant upregulation of adh, acs, ach1, and ackA directly related to the initial steps of acetate and furan aldehydes metabolisms. ZW70 strain can effectively remove the high concentration inhibitors cocktail from the hydrolysates derived from pretreated wheat straw and furfural residues. The biodetoxified hydrolysates by ZW70 were successfully used for cellulose chiral L-lactic acid production with the titers of ∼110 g/L, which were over 20 % higher than that detoxified by parental strain.

Chiral Bell-State Transfer via Dissipative Liouvillian Dynamics.

Chiral state transfer along closed loops in the vicinity of an exceptional point is one of the many counterintuitive observations in non-Hermitian physics. The application of this property beyond proof-of-principle in quantum physics, is an open question. In this work, we demonstrate chiral state conversion between singlet and triplet Bell states through fully quantum Liouvillian dynamics. Crucially, we demonstrate that this property can be used for the chiral production of Bell states from separable states with a high fidelity and for a large range of parameters. Additionally, we show that the removal of quantum jumps from the dynamics through postselection can result in near-perfect Bell states from initially separable states. Our work presents the first application of chiral state transfer in quantum information processing and demonstrates a novel way to control entangled states by means of dissipation engineering.

Chiral α-Aminophosphonates as Ligands in Copper-Catalyzed Asymmetric Oxidative Coupling of 2-Naphthols.

Chiral α-aminophosphonates with adjacent carbon and phosphonate stereogenic centers have been employed as ligands in the copper-catalyzed oxidative coupling of 2-naphthols, resulting in the production of chiral BINOLs in favorable yields and moderate to good enantiomeric excess. This represents the first application of chiral P-based ligands to enable such a transformation. The synthesis of these chiral α-aminophosphonate ligands offers a significant advantage over approaches that typically necessitate elaborate synthetic processes for chiral ligand production.

Enhanced catalytic efficiency for chiral alcohol pharmaceutical intermediate production using polyacrylic acid (PAA)-based nano-bienzyme conjugates at the organic-buffer biphasic interface

Nano-bienzyme conjugates (nano-BECs) for catalysing one-pot reactions can simplify the reaction process and improve the efficiency of biocatalytic reactions. In this work glucose dehydrogenase (GDH) and Candida parapsilosis secondary alcohol dehydrogenase (CpSADH) were immobilised with polyacrylic acid (PAA) to form two types of PAA-based nano-BECs (2-S, 2-P), which achieved in situ regeneration of NADH for the production of chiral alcohols in oil-water biphasic systems, such as the key chiral pharmaceutical intermediate of (S)-1-(3-trifluoromethylphenyl)ethanol ((S)-TMPE). At a substrate concentration of 100 mM 3-trifluoromethylacetophenone (TMAP), the conversion catalyzed by nano BEC was 98.2 % in dipentyl phthalate-phosphate buffer biphasic system. In contrast, the conversion was only 39.5 % and 38.9 % in the PB buffer and 8.4 % and 7.7 % in dipentyl phthalate system. The experiments on kinetic constants demonstrated that the catalytic efficiencies (Kcat/Km) of nano-BECs (2-S, 2-P) were enhanced by 6.7-fold and 7-fold, respectively, compared to free CpSADH. Additionally, nano-BECs showed stronger coenzyme regeneration ability. Under optimal reaction conditions, nano-BECs achieved 96.2 % conversion of TMAP to (S)-TMPE in an organic-buffer system with e.e. > 99.6 % at 300 mM substrate concentration. This research expands the potential applications of polymers and proteins, and offers valuable insights into the synergistic effects of PAA on bienzyme catalytic systems in biocatalysis.

Selective Stereoretention of Carbohydrates upon C-C Cleavage Enabling D-Glyceric Acid Production with High Optical Purity over a Ag/γ-Al2O3 Catalyst.

Chiral carboxylic acid production from renewable biomass by chemocatalysis is vitally important for reducing our carbon footprint, but remains underdeveloped. We herein establish a strategy that make use of a stereogenic center of biomass to achieve a rare example of D-glyceric acid production with the highest yield (86.8 %) reported to date as well as an excellent ee value (>99 %). Unlike traditional asymmetric catalysis, chiral catalysts/additives are not required. Ample experiments combined with quantum chemical calculations established the origins of the stereogenic center and catalyst performance. The chirality at C4 in D-xylose was proved to be retained and successfully delivered to C2 in D-glyceric acid during C-C cleavage. The remarkable cooperative-roles of Ag+ and Ag0 in the constructed Ag/γ-Al2O3 catalyst are disclosed as the crucial contributors. Ag+ was responsible for low-temperature activation of D-xylose, while Ag0 facilitated the generation of active O* from O2. Ag+ and active O* cooperatively promoted the precise cleavage of the C2-C3 bond, and more importantly O* allowed the immediate fast oxidization of the D-glyceraldehyde intermediate to stabilize D-glyceric acid, thereby inhibiting the side reaction that induced racemization. This strategy makes a significant breakthrough in overcoming the limitation of poor enantioselectivity in current chemocatalytic conversion of biomass.

Biocatalytic reductive aminations with NAD(P)H-dependent enzymes: enzyme discovery, engineering and synthetic applications.

Chiral amines are pivotal building blocks for the pharmaceutical industry. Asymmetric reductive amination is one of the most efficient and atom economic methodologies for the synthesis of optically active amines. Among the various strategies available, NAD(P)H-dependent amine dehydrogenases (AmDHs) and imine reductases (IREDs) are robust enzymes that are available from various sources and capable of utilizing a broad range of substrates with high activities and stereoselectivities. AmDHs and IREDs operate via similar mechanisms, both involving a carbinolamine intermediate followed by hydride transfer from the co-factor. In addition, both groups catalyze the formation of primary and secondary amines utilizing both organic and inorganic amine donors. In this review, we discuss advances in developing AmDHs and IREDs as biocatalysts and focus on evolutionary history, substrate scope and applications of the enzymes to provide an outlook on emerging industrial biotechnologies of chiral amine production.

Construction novel yeast surface-displaying glucose-leucine dehydrogenase fusions for cofactor recycling and chiral amino acid biosynthesis

Whole-cell biosynthesis of chiral amino acids (AAs) is of significant potential application. However, the difficulty of reactant translocation cell membranes limits their industrial utilization. In this study, a fusion enzyme integrating glucose dehydrogenase and leucine dehydrogenase (BmGDH-L-EsLDH) was engineered and anchored on the yeast surface, thereby enhancing cofactor recycling and catalytic efficiency of chiral AA production. In order to optimize the expression, a strategy of codon optimization was implemented. The surface-displayed opt-BmGDH-L-EsLDH reached to 2380 U/g (wet cell weight), which was 1.8-fold higher than the control. Moreover, the fusion enzyme markedly improved temperature and pH tolerance compared to the single enzyme. The NADH recycling efficiency of the fusion enzyme was augmented by 8-fold compared to the individual enzyme. The production of L-tert-leucine reached 2710 g/L/d in 200 mL scale without the addition of external NADH, achieving a total turnover number (TTN) of 9960 for NADH recycling. The surface-displayed BmGDH-L-EsLDH fusion demonstrated robust performance in biosynthesis of chiral amino acids.

Aqueous Circularly Polarized Luminescence Induced by Homopolypeptide Self-Assembly.

Remarkable advances have been achieved in solution self-assembly of polypeptides from the perspective of nanostructures, mechanisms, and applications. Despite the intrinsic chirality of polypeptides, the promising generation of aqueous circularly polarized luminescence (CPL) based on their self-assembly has been rarely reported due to the weak fluorescence of most polypeptides and the indeterminate self-assembly mechanism. Here, we propose a facile strategy for achieving aqueous CPL based on the self-assembly of simple homopolypeptides modified with a terminal group featuring both twisted intramolecular charge transfer and aggregation-induced emission properties. A morphology-dependent CPL can be observed under different self-assembly conditions by altering the solvents. A nanotoroid-dispersed aqueous solution with detectable CPL can be obtained by using tetrahydrofuran as a good solvent for the self-assembly, which is attributed to the involvement of the terminal group in the chiral environment formed by the homopolypeptide chains. However, such a chiral packing mode cannot be realized in nanorods self-assembled from dioxane, resulting in an inactive CPL phenomenon. Furthermore, CPL signals can be greatly amplified by co-assembly of homopolypeptides with the achiral small molecule derived from the terminal group. This work not only provides a pathway to construct aqueous CPL-active homopolypeptide nanomaterials but also reveals a potential mechanism in the self-assembly for chiral production, transfer, and amplification in polypeptide-based nanostructures.

Light Controlled Nanobiohybrids for Modulating Chiral Alcohol Synthesis.

The modulation of whole-cell activity presents a considerable challenge in biocatalysis. Conventional approaches to whole-cell catalysis, while having their strengths, often rely on complex and deliberate enzyme designs, which could result in difficulties in activity modulation and prolonged response times. Additionally, the activity of intracellular enzymes in whole-cell catalysis is influenced by temperature. To address these limitations, we introduced a relationally designed nanobiohybrid system that utilized light to modulate whole-cell catalysis for chiral alcohol production. By incorporating platinum nanoparticles onto Rhodotorula sp. cell surfaces, the nanobiohybrid capitalized on the photothermal properties of the nanoparticles to regulate the overall cell activity. When exposed to light, the Pt nanoparticles generate heat through the photothermal effect, consequently leading to an increase in the catalytic activity of the whole cells. This innovative approach facilitates control over whole-cell production and provides an efficient method for regulating biocatalytic processes. The findings of this study demonstrate the significant potential of switchable control strategies in biomanufacturing across a wide range of industries.

Stairway to Stereoisomers: Engineering Short- and Medium-Chain Ketoreductases To Produce Chiral Alcohols.

The short- and medium-chain dehydrogenase/reductase superfamilies are responsible for most chiral alcohol production in laboratories and industries. In nature, they participate in diverse roles such as detoxification, housekeeping, secondary metabolite production, and catalysis of several chemicals with commercial and environmental significance. As a result, they are used in industries to create biopolymers, active pharmaceutical intermediates (APIs), and are also used as components of modular enzymes like polyketide synthases for fabricating bioactive molecules. Consequently, random, semi-rational and rational engineering have helped transform these enzymes into product-oriented efficient catalysts. The rise of newer synthetic chemicals and their enantiopure counterparts has proved challenging, and engineering them has been the subject of numerous studies. However, they are frequently limited to the synthesis of a single chiral alcohol. The study attempts to defragment and describe hotspots of engineering short- and medium-chain dehydrogenases/reductases for the production of chiral synthons.

Recent Progress in the Mechanism and Engineering of α/β Hydrolases for Chiral Chemical Production

Chiral compounds are valuable industrial products and intermediates, and the production of chemicals with high enantiopurity is one of the major objects in asymmetric catalysis. Compared with traditional chemical synthesis, enzymatic synthesis can produce chiral molecules under sustainable conditions which are much greener, more economical, and more environmentally friendly. The superfamily of α/β hydrolases includes a lot of diverse enzymes showing excellent chemo-, regio-, and enantio-selectivity in asymmetric synthesis and many of them are biocatalysts in industry. This review outlines the current knowledge of the structures and reaction mechanism of α/β hydrolases and summarizes the screening and protein engineering efforts to develop biocatalysts for chiral chemicals production in recent years. Other strategies such as whole-cell catalysis and protein immobilization to improve the performance of α/β hydrolases are also discussed. The progress in biocatalyst development based on α/β hydrolases will promote the biosynthesis of chiral compounds, thus contributing to the green and sustainable development of the chemical and pharmaceutical industry.

Protein‐Mediated Biosynthesis of Semiconductor Nanocrystals for Photocatalytic NAD(P)H Regeneration and Chiral Amine Production

Protein‐Mediated Biosynthesis of Semiconductor Nanocrystals for Photocatalytic NAD(P)H Regeneration and Chiral Amine Production

Protein-Mediated Biosynthesis of Semiconductor Nanocrystals for Photocatalytic NAD(P)H Regeneration and Chiral Amine Production.

The use of predesigned bioengineered proteins for self-grown nanomaterials is a promising strategy that opens new scientific directions for biotic-abiotic nano-bio hybrid configurations. The unique properties of nanomaterials can alter the original biological paradigm to allow novel metabolic routes or new activation triggers. In this work, we present a synthetic methodology for self-grown cadmium sulfide quantum dots in a 12-mer bioengineered stable protein 1 under ambient conditions. The sized controlled crystalline QDs are characterized and utilized for NADPH regeneration that is in turn used for the activation of the imine reductase enzyme. The presented nano-bio hybrid system enables the production of a single enantiomeric product that is required for the pharmaceutical industry. Our designed system presents superior activity and can continuously operate for at least 22 hrs with 82 % conversion efficiency. The obtained results may lay the foundations for future nano-bio hybrid systems that can operate both in vitro and in vivo.

Solid-State Deracemization via Temperature Cycles in Continuous Operation: Model-Based Process Design.

Solid-state deracemization via temperature cycles converts a racemic crystal mixture into an enantiopure product by periodic cycling of the temperature in the presence of a racemization catalyst. A continuous counterpart of this conventional batch-operated process is proposed that can be performed in mixed suspension mixed product removal crystallizers (MSMPRCs). More specifically, three different configurations are described to perform periodic forcing via temperature cycles, which differ from each other in the type of the feed and in the withdrawal system. We have developed a model by extending our recent population balance equation model of batch solid-state deracemization via temperature cycles, and we exploit this tool to analyze the start-up and periodic steady-state behavior. Moreover, we compare the performance of the different configurations based on the selected key performance indicators, namely, average periodic steady-state enantiomeric excess and productivity. The process with solution feed yields pure enantiomers, while the solid and suspension-fed process alternatives result in highly enantiomerically enriched crystals. We further design an MSMPRC cascade to overcome this purity limitation. This work discusses guidelines on how to transform the batch process of temperature cycles into a continuous operation, which enables stable, unattended operation and chiral crystal production with consistent product quality.

Enhancing cofactor regeneration of cyanobacteria for the light-powered synthesis of chiral alcohols

Cyanobacteria Synechocystis sp. PCC 6803 was exploited as green cell factory for light-powered asymmetric synthesis of aromatic chiral alcohols. The effect of temperature, light, substrate and cell concentration on substrate conversions were investigated. Under the optimal condition, a series of chiral alcohols were synthesized with conversions up to 95% and enantiomer excess (ee) > 99%. We found that the addition of Na2S2O3 and Angeli’s Salt increased the NADPH content by 20% and 25%, respectively. As a result, the time to reach 95% substrate conversion was shortened by 12 h, which demonstrated that the NADPH regeneration and hence the reaction rates can be regulated in cyanobacteria. This blue-green algae based biocatalysis showed its potential for chiral compounds production in future.

Rational engineering of the Plasmodium falciparuml-lactate dehydrogenase loop involved in catalytic proton transfer to improve chiral 2-hydroxybutyric acid production

l-lactate dehydrogenases (LDHs) has been widely studied for their ability to reduce 2-keto acids for the production of 2-hydroxy acids, whereby 2-hydroxybutyric acids (2-HBA) is among the most important fundamental building blocks for synthesizing pharmaceuticals and biodegradable materials. However, LDHs usually show low activity towards 2-keto acids with longer side chain such as 2-oxobutyric acid (2-OBA). Here rational engineering of the Plasmodium falciparum LDH loop with residue involved in the catalytic proton transfer was initially studied. By combining homology alignment and structure-based design approach, we found that changing the charge characteristics or hydrogen bond network interactions of this loop could improve enzymatic catalytic activities and stabilities towards 2-OBA. Compared with wild type, variant N197Dldh showed 1.15 times higher activity and 2.73 times higher Kcat/Km. The half-life of variant N197Dldh at 40 °C increased to 77.9 h compared with 50.4 h of wild type. Furthermore, asymmetric synthesis of (S)-2-HBA with coenzyme regeneration revealed 95.8 g/L production titer within 12 h for variant N197Dldh, 2.05 times higher than using wild type. Our study indicated the importance of loop with residues involved in the catalytic proton transfer process, and the engineered LDH would be more suitable for (S)-2-HBA production.

Synthesis of (9,8) single-walled carbon nanotubes on CoSO4/SiO2 catalysts: The effect of Co mass loadings

Selective synthesis of single-walled carbon nanotubes (SWCNTs) with narrow chirality distribution at high production yield is critical to realize their applications in electronics and medicine. However, there is a significant trade-off between chirality selectivity and SWCNT yield. In this study, a series of CoSO4/SiO2 catalysts containing from 1 to 9 wt% of Co were prepared to systematically study how Co mass loadings affect the chirality selectivity and carbon production yield. SWCNTs were analyzed by spectroscopies and microscopes, while catalysts at three stages of SWCNT synthesis were studied by multiple characterization tools. Our results show that all CoSO4/SiO2 catalysts can grow (9,8) tubes. Specifically, the catalyst containing 3 wt% Co has the highest yield of (9,8) tubes, which grow from metallic Co particles with a size of around 1.2 nm. These Co particles are in-situ produced by reducing small Co9S8 particles of 2 nm in size, while unreduced Co9S8 particles help to stabilize metallic Co particles. High Co mass loadings lead to the formation of large Co and Co9S8 particles, which favor the formation of graphitic carbon and multi-walled carbon nanotubes. These findings provide useful insights to guide the development of novel catalysts for the chiral selective production of SWCNTs.

Rational design of halohydrin dehalogenase for efficient chiral epichlorohydrin production with high activity and enantioselectivity in aqueous-organic two-phase system

Halohydrin dehalogenases (HHDHs) represent as important biocatalysts for the production of valuable chiral epoxides and β-substituted alcohols. The HHDH from A. radiobacter (HheC) showed great potential for biosynthesis of (S)-epichlorohydrin ((S)-ECH). However, two bottlenecks including self-degradation and enzyme-mediated racemization of product in aqueous solution restricted its wide-spread applications. To solve the problems, the biocatalytic process was performed in aqueous-organic two-phase system and structure-based engineered of HheC was performed. A newly found zigzag tunnel connecting the active site region and outside space, as well as a depressed cave structure in formation of an extra three-pot herringbone channel were demonstrated to play a pivotal role affecting the activity and enantioselectivity of the enzyme. By mutation of the involved amino acid residues, two variants E255 T and E255 G showed significantly improved activity, with specific activity increased from 59.9 U/mg to 101.6 U/mg and 77.5 U/mg, respectively, while one variant Y93 L exhibited high enantioselectivity, with ee value of (S)-ECH maintained at >99 %. In addition, the synergic variant Y93 L/E255 T had both improved catalytic efficiency and enantioselectivity. The results not only facilitate the applicability of HheC for efficient production of (S)-ECH, but also give new insights into the structural-functional relationship of the enzyme.

Magnetic fields in heavy ion collisions: flow and charge transport

At the earliest times after a heavy-ion collision, the magnetic field created by the spectator nucleons will generate an extremely strong, albeit rapidly decreasing in time, magnetic field. The impact of this magnetic field may have detectable consequences, and is believed to drive anomalous transport effects like the Chiral Magnetic Effect (CME). We detail an exploratory study on the effects of a dynamical magnetic field on the hydrodynamic medium created in the collisions of two ultrarelativistic heavy-ions, using the framework of numerical ideal MagnetoHydroDynamics (MHD) with the ECHO-QGP code. In this study, we consider a magnetic field captured in a conducting medium, where the conductivity can receive contributions from the electromagnetic conductivity sigma and the chiral magnetic conductivity sigma _{chi }. We first study the elliptic flow of pions, which we show is relatively unchanged by the introduction of a magnetic field. However, by increasing the magnitude of the magnetic field, we find evidence for an enhancement of the elliptic flow in peripheral collisions. This effect is stronger at RHIC than the LHC, and it is evident already at intermediate collision centralities. Next, we explore the impact of the chiral magnetic conductivity on electric charges produced at the edges of the fireball. This initial sigma _chi can be understood as a long-wavelength effective description of chiral fermion production. We then demonstrate that this chiral charge, when transported by the MHD medium, produces a charge dipole perpendicular to the reaction plane which extends a few units in rapidity. Assuming charge conservation at the freeze-out surface, we show that the produced charge imbalance can have measurable effects on some experimental observables, like v_1 or langle sin phi rangle . This demonstrates the ability of a MHD fluid to transport the signature of the initial chiral magnetic fields to late times. We also comment on the limitations of the ideal MHD approximation and detail how further development of a dissipative-resistive model can provide a more realistic description of the QGP.

Baryogenesis from axion inflation

The coupling of an axion-like particle driving inflation to the Standard Model particle content through a Chern-Simons term generically sources a dual production of massless helical gauge fields and chiral fermions. We demonstrate that the interplay of these two components results in a highly predictive baryogenesis model, which requires no further ingredients beyond the Standard Model. If the helicity stored in the hyper magnetic field and the effective chemical potential induced by the chiral fermion production are large enough to avoid magnetic diffusion from the thermal plasma but small enough to sufficiently delay the chiral plasma instability, then the non-vanishing helicity survives until the electroweak phase transition and sources a net baryon asymmetry which is in excellent agreement with the observed value. If any of these two conditions is violated, the final baryon asymmetry vanishes. The observed baryon asymmetry can be reproduced if the energy scale of inflation is around Hinf ∼ 1010–1012 GeV with a moderate dependence on inflation model parameters.